# Medicinal Plants in the Prevention and Treatment of Colon Cancer

Authors: Paola Aiello; Maedeh Sharghi; Shabnam Malekpour Mansourkhani; Azam Pourabbasi Ardekan; Leila Jouybari; Nahid Daraei; Khadijeh Peiro; Sima Mohamadian; Mahdiyeh Rezaei; Mahdi Heidari; Ilaria Peluso; Fereshteh Ghorat; Anupam Bishayee; Wesam Kooti
Journal: Oxidative Medicine and Cellular Longevity
 (2019)

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## Abstract

The standard treatment for cancer is generally based on using cytotoxic drugs, radiotherapy, chemotherapy, and surgery. However, the use of traditional treatments has received attention in recent years. The aim of the present work was to provide an overview of medicinal plants effective on colon cancer with special emphasis on bioactive components and underlying mechanisms of action. Various literature databases, including Web of Science, PubMed, and Scopus, were used and English language articles were considered. Based on literature search, 172 experimental studies and 71 clinical cases on 190 plants were included. The results indicate that grape, soybean, green tea, garlic, olive, and pomegranate are the most effective plants against colon cancer. In these studies, fruits, seeds, leaves, and plant roots were used forin vitro and in vivo models. Various anticolon cancer mechanisms of these medicinal plants include induction of superoxide dismutase, reduction of DNA oxidation, induction of apoptosis by inducing a cell cycle arrest in S phase, reducing the expression of PI3K, P-Akt protein, and MMP as well; reduction of antiapoptotic Bcl-2 and Bcl-xL proteins, and decrease of proliferating cell nuclear antigen (PCNA), cyclin A, cyclin D1, cyclin B1 and cyclin E. Plant compounds also increase both the expression of the cell cycle inhibitors p53, p21, and p27, and the BAD, Bax, caspase 3, caspase 7, caspase 8, and caspase 9 proteins levels. In fact, purification of herbal compounds and demonstration of their efficacy in appropriate in vivo models, as well as clinical studies, may lead to alternative and effective ways of controlling and treating colon cancer.


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## Body

## 1. Introduction

An uncontrolled growth of the body’s cells can lead to cancer. Cancer of the large intestine (colon) is one of the main cause of death due to cancer. While the numbers for colon cancer are somewhat equal in women (47,820) and men (47,700), it will be diagnosed in (16,190) men (23,720) more than women. Multiple factors are involved in the development of colorectal cancer, such as lack of physical activity [1], excessive alcohol consumption [2], old age [3], family history [4], high-fat diets with no fiber and red meat, diabetes [5], and inflammatory bowel diseases, including ulcerative colitis and Crohn’s disease [6].Prevention of colorectal cancer usually depends on screening methods to diagnose adenomatous polyps which are precursor lesions to colon cancer [7]. The standard treatment for cancer is generally based on using cytotoxic drugs, radiotherapy, chemotherapy, and surgery [8]. Apart from these treatments, antiangiogenic agents are also used for the treatment and control of cancer progression [9].Colon cancer has several stages: 0, I, II, III, and IV. Treatment for stages 0 to III typically involves surgery, while for stage IV and the recurrent colon cancer both surgery and chemotherapy are the options [10]. Depending on the cancer stage and the patient characteristics, several chemotherapeutic drugs and diets have been recommended for the management of colorectal cancer. Drugs such as 5-fluorouracil (5-FU), at the base of the neoadjuvant therapies folfox and folfiri, are used together with bevacizumab, panitumumab, or cetuximab [7].Chemotherapy works on active cells (live cells), such as cancerous ones, which grow and divide more rapidly than other cells. But some healthy cells are active too, including blood, gastrointestinal tract, and hair follicle ones. Side effects of chemotherapy occur when healthy cells are damaged. Among these side effects, fatigue, headache, muscle pain, stomach pain, diarrhea and vomiting, sore throat, blood abnormalities, constipation, damage to the nervous system, memory problems, loss of appetite, and hair loss can be mentioned [11].Throughout the world, early diagnosis and treatment of cancer usually increase the individual’s chances of survival. But in developing countries, access to effective and modern diagnostic methods and facilities is usually limited for most people, especially in rural areas [12]. Accordingly, the World Health Organization (WHO) has estimated that about 80% of the world population use traditional treatments [13]. One of these treatments is phytotherapy, also known as phytomedicine, namely, the use of plants or a mixture of plant extracts for the treatment of diseases. The use of medicinal plants can restore the body’s ability to protect, regulate, and heal itself, promoting a physical, mental, and emotional well-being [14–16]. Various studies have shown the therapeutic effects of plants on fertility and infertility [17], hormonal disorders, hyperlipidemia [18], liver diseases [19], anemia [20], renal diseases [21], and neurological and psychiatric diseases [22]. Therefore, due to all the positive effects showed by medicinal plants, their potential use in cancer prevention and therapy has been widely suggested [23–25].Since the current treatments usually have side effects, plants and their extracts can be useful in the treatment of colon cancer with fewer side effects. The aims of this review are to present and analyse the evidence of medicinal plants effective on colon cancer, to investigate and identify the most important compounds present in these plant extracts, and to decipher underlying molecular mechanisms of action.


## 2. Literature Search Methodology

This is a narrative review of all research (English full text or abstract) studies conducted on effective medicinal plants in the treatment or prevention of colon cancer throughout the world. Keywords, including colon cancer, extract, herbs, plant extracts, and plants, were searched separately or combined in various literature databases, such as Web of Science, PubMed, and Scopus. Only English language articles published until July 2018 were considered.In the current narrative review, studies (published papers) were accepted on the basis of inclusion and exclusion criteria. The inclusion criterion was English language studies, which demonstrated an effective use of whole plants or herbal ingredients, as well as studies which included standard laboratory tests.In vivo and in vitro studies that were published as original articles or short communications were also included. The exclusion criteria included irrelevancy of the studies to the subject matter, not sufficient data in the study, studies on mushrooms or algae, and the lack of access to the full text. Reviews, case reports/case series, and letters to editors were also excluded but used to find appropriate primary literature.The abstracts of the studies were reviewed independently by two reviewers (authors of this study) on the basis of the inclusion and exclusion criteria. In case of any inconsistency, both authors reviewed the results together and solved the discrepancy. Data extracted from various articles were included in the study and entered into a check list after the quality was confirmed. This check list included some information: authors’ name, year of publication, experimental model, type of extract and its concentration or dose, main components, and mechanisms of action (if reported).


## 3. Results

### 3.1. Medicinal Plants and Colon Cancer

Overall, 1,150 articles were collected in the first step and unrelated articles were excluded later on according to title and abstract evaluation. Moreover, articles that did not have complete data along with congress and conference proceedings were excluded. Accordingly, a total of 1,012 articles were excluded in this step. Finally, 190 articles fulfilled the criteria and were included in this review. These papers were published within 2000-2017. A total of 190 plants were included in this study. Based on literature search, 172 experimental studies and 71 clinical cases were included.Overall, results indicate that grape, soybean, green tea, garlic, olive, and pomegranate are the most effective plants against colon cancer. In these studies, fruits, seeds, leaves, and plant roots were used forin vitro and in vivo studies.


#### 3.1.1.In Vitro Studies

Out of 172 studies, 75 were carried out on HT-29, 60 on HCT116, and 24 on Caco-2 cells (Table1). On HT-29 cells, both Allium sativum root extracts and Camellia sinensis leaf extracts induced cell apoptosis by two different mechanisms, respectively. In fact, the former showed inhibition of the PI3K/Akt pathway, upregulation of PTEN, and downregulation of Akt and p-Akt expression, while the latter was involved in attenuation of COX-2 expression and modulation of NFκB, AP-1, CREB, and/or NF-IL-6. Moreover, an antiproliferative activity has also been detected in Olea europaea fruit extracts, which increased caspase 3-like activity and were involved in the production of superoxide anions in mitochondria. An antiproliferative activity, by means of a blockage in the G2/M phase, has also been reported in Caco-2 cells by Vitis vinifera fruit extracts. Concerning HCT116 cells, several plants, such as American ginseng and Hibiscus cannabinus, induced cell cycle arrest in different checkpoints.Table 1
Cytotoxic effects of medicinal plants on colon cancer inin vitro models.
Scientific name
Parts used
Cell line
Conc.
Type of extract
Important compounds
Cellular effect
Mechanisms
References
Vitis vinifera
Fruit
HCT116
NM
Lyophilized
Hydroxycinnamic acids, proanthocyanidins, stilbenoids
Increase of dihydroceramides, sphingolipid mediators involved in cell cycle arrest, and reduction of the proliferation rate
(i) Increase of p53 and p21 cell cycle gate keepers(ii) Activation of the transcriptional factor Nrf2
[26, 27]
Fruit
Caco-2
365 mg/g
Methanolic
Catechin, epicatechin, quercetin, gallic acid
Antiproliferative activity and direct initiation of cell death
Blockage in the G2/M phase
[28, 29]
Seed
Caco-2
10–25μg/mL
Aqueous
Procyanidins
(i) Increased crypt depth(ii) Inhibited cell viability and decreased histological damage score
Reduced MPO (myeloperoxidase) activity
[29]
Skin
NM
7.5, 30, 60μg/mL
Methanolic
4′-Geranyloxyferulic acid
NM
NM
[30]
Seed
Colon cancer stem cells
6.25, 12.5, 25μg/mL
NM
(+)-catechin, (−)-epicatechin
NM
(i) Increment of p53, Bax/Bcl-2 ratio, and cleaved PARP(ii) Inhibition of Wnt/β-catenin signaling
[31]
Allium sativum
Root
HT-29
20, 50, 100 mg/mL
Ethanolic
NM
Induction of apoptosis and cell cycle arrest
(i) Inhibition of the PI3K̸Akt pathway(ii) Upregulation of PTEN and downregulation of Akt and p-Akt expression
[32]
Glycine max
Seed
Caco-2, SW620, HT-29
12.5μg/mL
Aqueous
Anthoxanthin
Cell death and significant reduction of cell density
Enhancement of Rab6 protein levels
[33]
Seed
HT-29
240, 600 ppm
Crude
Saponin
Suppression of PKC activation and increase of alkaline phosphatase activity
[33]
Seed
HT-29
NM
Crude
Saponin
NM
(i) Suppression of the degradation of IκBα in PMA-stimulated cells(ii) Downregulation of COX-2 and PKC expressions
[34]
Camellia sinensis
Leaf
HT-29
0, 10, 30, 50μM
Aqueous
Catechin, epigallocatechin gallate
1.9-fold increase in tumor cell apoptosis and a 3-fold increase in endothelial cell apoptosis
(i) Suppression of ERK-1 and ERK-2 activation(ii) Suppression of VEGF expression
[35]
Leaf
Caco-2, HT-29
300μM
Aqueous
Theaflavins (TF-2T, F-3, TF-1)
Human colon cancer cell apoptosis induction
Modulation of NFκB, AP-1, CREB, and/or IL-6
[36]
Leaf
HT-29
68-800.73 μg/mL
Hot water extract
Flavan-3-ol (catechin & tannin) & polyphenols (teadenol B)
Inhibition of proliferation of HT-29 cells
Increased expression levels of caspases 3/7, 8, and 9
[35]
Olea europaea
Fruit
HT-29
150, 55.5200 and 74 μmol/L
Methanolic and chloroform
Maslinic acid, oleanolic acid
Antiproliferative activity
(i) Increased caspase 3-like activity to 6-fold(ii) Production of superoxide anions in the mitochondria
[37]
Fruit, leaf
SW480 and HT-29
100–400 m/z
Methanolic & hexane
Oleic acid, linoleic acid, gamma-linolenic acid, lignans, flavonoids, secoiridoids
Reduced cell growth in both cell lines
(i) Limited G2M cell cycle(ii) Depressed cyclooxygenase-2 expression in HT-29 cells(iii) Suppression of β-catenin/TCF signaling in SW480 cells(iv) Promotion of the entry into subG1 phase
[38]
Fruit
Caco-2
50μM
Aqueous
Phenolic compounds, authentic hydroxyl tyrosol (HT)
Reduced proliferation of Caco-2 cells
Reduction of the methylation levels of CNR1 promoter
[39]
Fruit
HT115
25μg/mL
Hydroethanolic
Phenolic compounds (p-hydroxyphenyl ethanol, pinoresinol & dihydroxyphenyl ethanol)
NM
Inhibition by reduced expression of a range ofα5 & β1
[40]
Olive mill wastewater
HT-29, HCT116, CT26
NM
Methanolic
Hydroxytyrosol
(i) Inhibited proliferation(ii) Inhibited migration and invasion
(i) Reduced sprout formation(ii) Inhibited VEGF and IL-8 levels
[41]
Fruit
Caco-2
0-2,000μg/mL
Ethanolic
Tyrosol, hydroxytyrosol, oleuropein, rutin, quercetin and glucoside forms of luteolin and apigenin
NM
(i) Induction of the cell cycle arrest in S-phase
[42]
Punica granatum
Juice
HT-29
50 mg/L
Aqueous
Ellagitannins, punicalagin
Inhibition of cancer cell proliferation
(i) Suppressed TNFR-induced COX-2 protein expression(ii) Reduced phosphorylation of the p65 subunit and binding to the NFκB response element
[43]
Seed
LS174
63.2μg/mL
Supercritical fluid
Punicic acid,γ-tocopherol, α-tocopherol
Cytotoxic activity
(i) Slightly decreased development of tubules from elongated cell bodies(ii) Reduction of the number of cell connections
[44]
Glycyrrhiza glabra
Root
HT-29
12.2 and 31μg/mL
Ethanolic
Licochalcone
NM
Increase of the protein levels of proapoptotic Bax
[37]
Opuntia ficus-indica
Fruit
Caco-2
115μM
Aqueous
Betalain pigment indicaxanthin
Apoptosis of proliferating cells
(i) Demethylation of the tumor suppressor p16INK4a gene promoter(ii) Reactivation of the silenced mRNA expression and accumulation of p16INK4a
[38]
Fruit
HT-29 & Caco-2 & NIH 3 T3 (as control)
Against HT-29 (4.9μg/mL) against Caco-2 (8.2 μg/mL)
Alkaline hydrolysis with NaOH
Isorhamnetin glycosides (IG5 and IG6)-phenol
Cell death through apoptosis and necrosis
Increased activity of caspase 3/7
[45]
Piper betle
Leaf
HT-29 and HCT116
200.0μg/mL
Aqueous
Hydroxychavicol
Antioxidant capacity and induction of a greater apoptotic effect
(i) Scavenging activity(ii) Formation of electrophilic metabolites
[41, 46]
Fragaria×ananassa
Fruit
HT-29
0.025, 0.05, 0.25, 0.5%
Ethanolic
Ascorbate, ellagic acid
Decreased proliferation of HT-29 cells
Increase in the levels of 8OHA and decrease in the levels of 8OHG
[40]
Sasa quelpaertensis
Leaf
HT-29 HCT116
0, 100, 200, 300 mg/L
Ethanolic
p-Coumaric acid, tricin
Inhibited colony formation
Nonadherent sphere formation suppressed CD133+ & CD44+ population
[41]
Salvia chinensis
Stem
HCT116, COLO 205
10, 20, 40,60, 80, 100 mg/L
Polyphenolic
Terpenoids, phenolic acid, flavonoids, dibenzylcyclooctadiene
Apoptosis & loss of mitochondrial membrane
Induced G0/G1 cell cycle
[42]
Rubus idaeus L.
Fruit
HT-29, HT-115, Caco-2
3.125, 6.25, 12.5, 25, 50 mg/L
Acetate
Polyphenol, anthocyanin, ellagitannin
NM
Decreased population of cells in G1 phase
[47]
LoVo
50μL
Aqueous
NM
Inhibited proliferation of LoVo
Suppression of the NFκB pathway
[48]
Curcuma longa
Root
HT-29, HCT15, DLD1, HCT116
(i) Short-term assay: four 10-fold dilutions (100 to 0.1 mg/L)(ii) Long-term assay: 5, 10, 20 mg/L
Ethanolic
Curcumin (diferuloylmethane)
Inhibited formation of HCT116 spheroids
NM
[49]
Eleutherococcus senticosus
Root
HCT116
12.5, 25, 50, 100
Methanolic
Eleutherosides, triterpenoid saponins, glycans
NM
Activation of natural killer cells and thus enhancement of immune function
[50]
Tabernaemontana divaricata L.
Leaf
HT-29, HCT15
10, 30, 100 mg/L
Ethyl acetate, chloroform, methanolic
Alkaloids
NM
Inhibited the unwinding of supercoiled DNA
[45]
Millingtonia hortensis
Root, flower, leaf
RKO
50, 100, 200, 400, 800 mg/L
Aqueous
Phenylethanoid glycoside, squalene, salidroside, 2-phenyl rutinoside
Apoptosis induction
(i) Increase of fragmented DNA(ii) Decrease of the expression of antiapoptotic proteins, Bcl-xL and p-BAD
[46]
Powder
RKO
200, 400, 800μg/mL
Aqueous
Water soluble compounds
Antiproliferative effect
NM
[51]
Thai purple rice
Seed
Caco-2, Cat. No. HTB-37
16.11μg/mL
Methanol acidified
Cyanidin-3-glucoside and peonidin-3-glucoside, anthocyanins, phenolic compounds
(i) Antioxidation of anthocyanins and phenols(ii) Antiproliferation of colon cancer cells
NM
[52]
Annona muricata
Leaf
HCT116, HT-29
11.43±1.87μg/ml and 8.98±1.24μg/ml
Ethanolic
Alkaloids, acetogenins, essential oils
Block of the migration and invasion of HT-29 and HCT116 cells
(i) Cell cycle arrest at G1 phase(ii) Disruption of MMP, cytochrome c leakage and activation
[53]
NM
HT-29, HCT116
<4, <20μg/mL
EtOAc
Annopentocin A, annopentocin B, annopentocin C, cis- and trans-annomuricin-D-ones, annomuricin E
NM
Suppression of ATP production and NADH oxidase in cancer cells
[54]
Pistacia lentiscus L. var. chia
Leaf
HCT116
NM
Ethanolic
Resin, known as Chios mastic gum (CMG)
Causes several morphological changes typical of apoptosis in cell organelles
(i) Induction of cell cycle arrest at G1 phase(ii) Activation of pro-caspases 8, 9, 3
[55]
Resin
HCT116
100μg/mL
Hexane
Caryophyllene
Induction of the anoikis form of apoptosis in human colon cancer HCT116 cells
(i) Induction of G1 phase arrest(ii) Loss of adhesion to the substrate
[56]
American ginseng (Panax quinquefolius)
Biological constituents
HCT116
0-2.0 mg/mL
Aqueous
Ginseng (GE) or its ginsenoside (GF) and polysaccharide (PS)
Proliferation was inhibited by GE, GF, and PS in wild-type and p21 cells
(i) Cells arrest in G0/G1 phase and increment of p53 and p21 proteins(ii) Increment of Bax and caspase 3 proteins expression
[57]
Purple-fleshed potatoes
Fruit
Colon cancer stem cells
5.0μg/mL
Ethanol, methanol, ethyl acetate
Anthocyanin,β-catenin, cytochrome c
Critical regulator of CSC proliferation and its downstream proteins (c-Myc and cyclin D1) and elevated Bax and cytochrome c
(i) Cytochrome c levels were elevated regardless of p53 status(ii) Mitochondria-mediated apoptotic pathway(iii) Suppressed levels of cytoplasmic and nuclear β-catenin
[58]
Phaseolus vulgaris
Leaf
HT-29
NM
Ethanolic
Polysaccharides, oligosaccharides
Changes in genes involved or linked to cell cycle arrest
(i) Inactivation of the retinoblastoma phosphoprotein(ii) Induction of G1 arrest(iii) Suppression of NF-jB1(iv) Increase in EGR1 expression
[59]
Opuntia spp.
Fruit
HT-29
5.8±1.0, 7.5±2.0, 12±1% (V/V)
Hydroalcoholic
Betacyanins, flavonoids (isorhamnetin derivatives) and phenolic acids (ferulic acid)
NM
Induced cell cycle arrest at different checkpoints—G1, G2/M, and S
[60]
Suillus luteus
NM
HCT15
400μg/mL
Methanolic
Protocatechuic acid, cinnamic acid,α-tocopherol, β-tocopherol, mannitol, trehalose, polyunsaturated fatty acids, monounsaturated fatty acids, saturated fatty acids
(i) Increase in the cellular levels of p-H2A.X, which is suggestive of DNA damage
(i) Inhibition of cell proliferation in G1 phase(ii) Increase in the cellular levels of p-H2A.X
[61]
Poncirus trifoliata
Leaf
HT-29
0.63μM
Aqueous (in acetone)
β-Sitosterol, 2-hydroxy-1,2,3-propanetricarboxylic acid 2-methyl ester
Arrest of cell growth was observed withβ-sitosterol
NM
[62]
Rosmarinus officinalis L.
Leaf
SW 620, DLD-1
0-120μg/mL
Methanolic
Polyphenols
Antiproliferative effect of 5-FU
Downregulation of TYMS and TK1, enzymes related to 5-FU resistance
[63]
Leaf
HT-29
SC-RE 30μg/mL and CA 12.5 μg/mL
Ethanolic
Polyphenols (carnosic acid (CA) and carnosol)
(i) Upregulation of VLDLR gene as the principal contributor to the observed cholesterol accumulation in SC-RE-treated cells(ii) Downregulation of several genes involved in G1-S
Activation of Nrf2 transcription factor and common regulators, such as XBP1 (Xbp1) gene related to the unfolded protein response (UPR)
[64]
NM
HT-29
10, 20, 30, 40, 50, 60, 70μg/mL
NM
Carnosic acid, carnosol, rosmarinic acid, rosmanol
NM
NM
[65]
Leaf
HGUE-C-1, HT-29, and SW480
20–40 mg/mL
CO2-supercritical fluid extract
Carnosic acid, carnosol, and betulinic acid
NM
(i) Prooxidative capability by increasing the intracellular generation of ROS(ii) Activation of Nrf2
[66]
Glehnia littoralis
Leaf
HT-29
50 mg/mL
Methanolic
Bergapten, isoimpinellin, xanthotoxin, imperatorin, panaxydiol, falcarindiol, falcarinol
Induced apoptosis by the decreased expression of the antiapoptotic Bcl-2 mRNA
(i) Reduced expression of Bcl-2(ii) Reduced expression levels of iNOS and COX-2
[67]
Verbena officinalis
Leaf
HCT116
20 mg/mL
Aqueous
Phenylethanoid glycosides, diacetyl-O-isoverbascoside, diacetyl-O-betonyoside A, and diacetyl-O-betonyoside A
(i) Substantial tumor cell growth inhibitory activity(ii) Time-dependent cytotoxicity against both cell lines
(i) Increased lipophilicity of molecules seemed to be responsible for enhanced cytotoxicity(ii) Antiproliferative activity is determined by the number of acetyl groups and also by their position in the aliphatic rings
[68]
Mentha spicata
Leaf
RCM-1
12.5μg/mL
N-Hexane
Acetic acid 3-methylthio propyl ester (AMTP), methyl thio propionic acid ethyl ester (MTPE)
Exhibited antimutagenic activity
Auraptene (7-geranyloxycoumarin) having a monoterpene moiety andβ-cryptoxanthin (one of the tetraterpenes) increased antibody production
[69]
Euphoria longana Lam.
Seed
SW 480
0–100μg/mL
Ethanolic
Corilagin, gallic acid, ellagic acid
(i) Antiangiogenetic properties(ii) All fractions showed the anti-VEGF secretion activity
Release and expression of VEGF indicated that all fractions showed the anti-VEGF secretion activity
[70]
Sutherlandia frutescens
Flower
Caco-2
1/50 dilution of the ethanolic extract
Ethanolic
Amino acids, including L-arginine and L-canavanine, pinitol, flavonoids, and triterpenoid saponins as well as hexadecanoic acid andγ-sitosterol
Disruption of the key molecules in the PI3K pathway thereby inducing apoptosis
Decrease in cell viability and increment in pyknosis as well as loss in cellular membrane integrity
[71]
Melissa officinalis
Leaf
HT-29, T84
346, 120μg/mL
Ethanolic
Phenolic acids (rosmarinic acid, coumaric acid, caffeic acid, protocatechuic acid, ferulic acid, chlorogenic acid), flavonoids, sesquiterpenes, monoterpenes, triterpenes
(i) Inhibited proliferation of colon carcinoma cells(ii) Induced apoptosis through formation of ROS
(i) G2/M cell cycle arrest(ii) Cleavage of caspases 3 and 7(iii) Induced phosphatidylserine externalization in colon carcinoma cells(iv) Induced formation of ROS in colon carcinoma cells
[72]
Sargassum cristaefolium
Leaf
HT-29
500 mg/mL
Ethanolic
Fucoidans
(i) Reduction of free radicals(ii) DPPH radical scavenging
Accumulation of cells in G0/G1 phase
[73]
Hedyotis diffusa
NM
HT-29
400 mg/mL
Ethanolic and then DMSO
Octadecyl (E)-p-coumarate, P-E-methoxy-cinnamic acid, ferulic acid, scopoletin, succinic acid, aurantiamide acetate, rubiadin
Suppress tumor cell growth and induce the apoptosis of human CRC cells
(i) Block G1/S progression(ii) Induce the activation of caspases 9 and 3(iii) Inhibit IL-6-mediated STAT3 activation(iv) Downregulate the mRNA and protein expression levels of cyclin D1, CDK4, Bcl-1, and Bax
[74]
Zingiber officinale Roscoe
Peel
LoVo
100 mg/mL
Ethanolic
Linoleic acid methyl ester,α-zingiberene, and zingiberone
Interesting antiproliferative activity against colorectal carcinoma
NM
[75]
Scutellaria barbata
Leaf
LoVo
413.3 mg/L
Methanolic
Scutellarein, scutellarin, carthamidin, isocarthamidin, wogonin
Induce cell death in the human colon cancer cell line
Increase in the sub-G1 phase and inhibition of cell growth
[76]
Pistacia atlantica, Pistacia lentiscus
Resin
HCT116
100μg/mL
Hexane extract
Caryophyllene
Induce the anoikis form of apoptosis in human colon cancer HCT116 cells
(i) Induce G1 phase arrest(ii) Loss of adhesion to the substrate
[56]
Citrus reticulata
Peel
SNU-C4
100μg/mL
Methanolic
Limonene, geranial, neral, geranyl acetate, geraniol,β-caryophyllene, nerol, neryl acetate
Induce the apoptosis on SNU-C4, human colon cancer cells
Expression of proapoptotic gene, Bax, and major apoptotic gene, caspase 3
[77]
Echinacea pallida, Echinacea angustifolia, Echinacea purpurea
Root
COLO320
150 mg/mL
Hexanic
Caffeic acid derivatives, alkylamides, polyacetylenes, polysaccharides
Induce apoptosis and promote nuclear DNA fragmentation
(i) Induce apoptosis by increasing caspase 3/7 activity(ii) Promote nuclear DNA fragmentation
[78]
Nasturtium officinale
Leaf
HT-29
50μL/mL
Methanolic
Phenethyl isothiocyanate, 7-methylsulfinylheptyl, 8-methylsulfinyl
(i) Inhibition of initiation, proliferation, and metastasis
(i) Inhibited DNA damage(ii) Accumulation of cells in S phase of the cell cycle
[79]
Polysiphonia
NM
SW480, HCT15, HCT116, DLD-1
20 and 40μg/mL
Methanolic
2,5-Dibromo-3,4-dihydroxybenzyl n-propyl ether
Potentially could be used as a chemopreventive agent against colon cancer
(i) Inhibited Wnt/β-catenin pathway(ii) Repressed CRT in colon cancer cells(iii) Downregulated cyclin D1(iv) Activated the NFκB pathway
[80]
Aristolochia debilis Sieb. et Zucc.
Stem
HT-29
200μg/mL
Methanolic
Aristolochic acid, nitrophenanthrene carboxylic acids
Inhibition of proliferation and induction of apoptosis in HT-29 cells
(i) Induction of sub-G1 cell cycle(ii) Generation of ROS and decrease of the MMP(iii) Bax overexpression and increase of Bax/Bcl-2 ratio
[81]
Myrtaceae
Leaf
HCT116
100μg/mL (in vitro), 200 and 100 μg/disc (in vivo)
Methanolic
Phenols, flavonoid, betulinic acid
Strong inhibition of microvessel outgrowth
(i) Inhibition of tube formation on Matrigel matrix(ii) Inhibition of HUVECS migration (in vitro)(iii) Decreased nutrient and oxygen supply
[82]
Spica prunellae
Leaf
HT-29
200 mg/mL (in vitro), 600 mg/mL (in vivo)
Ethanolic
Rosmarinic acid
Inhibits CRC cell growth
(i) Suppresses STAT3 phosphorylation(ii) Regulates the expression of Bcl-2, Bax, cyclin D1, CDK4, VEGF-A, and VEGFR-2
[83]
Phytolacca americana
Root
HCT116
3200μg/mL
Ethanolic
Jaligonic acids, kaempferol, quercetin, quercetin 3-glucoside, isoquercitrin, ferulic acid
Control of growth and spread of cancer cells
Reduction in the expressions of MYC, PLAU, and TEK
[84]
Morus alba
Leaf
HCT15
13.8μg/mL
Methanolic
Epicatechin, myricetin, quercetin hydrate, luteolin, kaempferol, ascorbic acid, gallic acid, pelargonidine, p-coumaric acid
Cytotoxic effect on human colon cancer cells (HCT15)
(i) Apoptosis induction also involved in the downregulation of iNOS(ii) Fragmentation of DNA(iii) Upregulation of caspase 3 activity
[85]
Rhodiola imbricata
Leaf
HT-29
200μg/mL
Acetone and methanolic
Phenols, tannins, and flavonoids
(i) Antioxidant activity(ii) Inhibited proliferation of HT-29 cells
(i) Scavenge free radicals(ii) DPPH radical scavenging activity(iii) Increased metal chelating activity
[86]
Asiasarum heterotropoides F.
DriedA. radix
HCT116
20 mg/mL
Ethanolic
Asarinin and xanthoxylol
Inhibition of the growth of HCT116 cells
(i) Caspase-dependent apoptosis(ii) Regulation of p53 expression at transcription level
[87]
Podocarpus elatus
Fruit
HT-29
500 mg/mL
Methanolic
Phenolic and anthocyanin
Reduction of proliferation of colon cancer cells
(i) Cell cycle delay in S phase(ii) 93% downregulation of telomerase activity and decrease in telomere length(iii) Induced morphological alterations to HT-29 cells
[88]
Echinacea purpurea
Flower
Caco-2, HCT116
0–2,000 mg/mL
Hydroethanolic
Cichoric acid
(i) Inhibition of proliferation(ii) Decreased telomerase activity in HCT116 cells
(i) Decreased telomerase activity(ii) Activation of caspase 9(iii) Cleavage of PARP(iv) Downregulation of β-catenin
[89]
Root
COLO320
150 mg/mL
Hexanic
Caffeic acid derivatives, alkylamides, polyacetylenes, polysaccharides
Induce apoptosis by increasing significantly caspase 3/7 activity and promote nuclear DNA fragmentation
(i) Increase significantly caspase 3/7 activity(ii) Promote nuclear DNA fragmentation
[78]
Hop (Humulus lupulus L.), Franseria artemisioides
Leaf
NM
100 mg/kg b.w./day
Aqueous
Coumarin, lignans, quinones
30% reduction of tumor-induced neovascularization
NM
[90]
NM
Caco-2
NM
Ethanolic
Phenolic compounds, flavonoid, diterpenes
Digestive, gastroprotective, antiseptic, anti-inflammatory, and antiproliferative activity
NM
[91]
Fruit
NL-17
0, 50, 100, 150μg/mL
Methanolic
α-Mangostin (xanthone)
NM
(i) Induction of caspase 3 and caspase 9 activation(ii) Induced cell cycle arrest at G1/G0 phase
[92]
Stem, bark
HT-29
50μg/mL
Chloroform-soluble
β-Mangostin, garcinone D, cratoxyxanthone
Cytotoxic activity against HT-29 human colon cancer
Inhibition of p50 and p65 activation
[93]
Annona squamosa Linn
Leaf
HCT116
8.98μg/mL
Crude, Aq ethyl acetate
Acetogenins (annoreticuin & isoannoreticuin) and alkaloids dopamine, salsolinol, and coclaurine
Inhibition of growth and proliferation of tumor cells
(i) Reactive oxygen species (ROS) formation, lactate dehydrogenase (LDH) release(ii) Activation of caspases 3/7, 8, and 9
[94]
Derris scandens
Stem
HT-29
5-15μg/mL
Ethanolic
Benzyls and isoflavones (genistein, coumarins, scandinone)
Apoptosis and mitotic catastrophe of human colon cancer HT-29 cells
(i) Inhibition ofα-glucosidase activity(ii) Scavenge free radicals
[95]
Eupatorium cannabinum
Aerial parts
HT-29
25μg/mL
Ethanolic
Pyrrolizidine alkaloids (senecionine, senkirkine, monocrotaline, echimidine)
Induced alteration of colony morphology
(i) Upregulation of p21 and downregulation of NCL, FOS, and AURKA(ii) Mitotic disruption and nonapoptotic cell death via upregulation of Bcl-xL, limited TUNEL labeling, and nuclear size increase
[96]
Sorghum bicolor
The dermal layer of stalk
HCT116 & colon cancer stem cells
>16 and 103μg/mL
Phenolic-rich ethanolic, acetone
Apigeninidin & luteolinidin
Antiproliferative
Target p53-dependent and p53-independent pathways
[97]
Dermal and seed head
CCSC
NM
Methanolic
Apigeninidin, luteolinidin, malvidin 3-O-glucoside, apigenin, luteolin, naringenin, naringenin 7-O-glucoside, eriodictyol 5-glucoside, taxifolin, catechins
NM
(i) Elevation of caspase 3/7 activity(ii) Decrease in β-catenin, cyclin D1, c-Myc, and survivin protein levels(iii) Suppression of Wnt/β-catenin signaling in a p53-dependent (dermal layer) and partial p53-dependent (seed head) manner
[98]
Hibiscus cannabinus
Seed
HCT116
KSE (15.625μg/mL to 1,000 μg/mL)
Ethanolic
Gallic acid, p-hydroxybenzoic acid, caffeic acid, vanillic acid, syringic acid, and p-coumaric and ferulic acids
Cytotoxic activity against human colon cancer HCT116 cells
Apoptosis via blockade of mid G1-late G1-S transition thereby causing G1 phase cell cycle arrest
[99]
Salix aegyptiaca L.
Bark
HCT116 & HT-29
300μg/mL
Ethanolic
Catechin, salicin, catechol and smaller amounts of gallic acid, epigallocatechin gallate (EGCG), quercetin, coumaric acid, rutin, syringic acid, and vanillin
Anticarcinogenic effects in colon cancer cells
Apoptosis via inhibition of phosphatidylinositol 3-kinase/protein kinase B and mitogen-activated protein kinase signaling pathways
[100]
Rubus coreanum
Fruit
HT-29
400μg/mL
Aqueous
Polyphenols, gallic acid, sanguine
Induction of apoptosis
(i) Induced activity of caspases 3, 7, and 9(ii) Cleavage of poly(adenosine diphosphate-ribose) polymerase
[101]
Codonopsis lanceolata
Root
HT-29
200μg/mL
N-Butanol fraction
Tannins, saponins, polyphenolics, alkaloids
Apoptosis in human colon tumor HT-29 cells
(i) Induced G0/G1 arrest(ii) Enhancement of expression of caspase 3 and p53 and of the Bax/Bcl-2 ratio
[102]
Gleditsia sinensis
Thorn
HCT116
800μg/mL
Aqueous
Flavonoid, lupine acid, ellagic acid glycosides
(i) Increase in p53 levels(ii) Downregulation of the checkpoint proteins, cyclin B1, Cdc2, and Cdc25c
Inhibition of proliferation of colon cancer cells
[90]
Thorn
HCT116
600μg/mL
Ethanolic
NM
Inhibitory effect on proliferation of human colon cancer HCT116 cells
(i) Caused cell cycle arrest at G2/M phase together with a decrease of cyclin B1 and Cdc2(ii) Progression from G2/M phase
[91]
Ligustrum lucidum
Fruit
DLD-1
50μg/mL
Aqueous
Oleanolic acid, ursolic acid
Inhibited proliferation
(i) Reduction of Tbx3 rescued the dysregulated P14ARF-P53 signaling
[94]
Zingiber officinale
Rhizome
HCT116
5μM
Ethanolic
6-Paradol, 6- and 10-dehydrogingerdione, 6- and 10-gingerdione, 4-, 6-, 8-, and 10-gingerdiol, 6-methylgingerdiol, zingerone, 6-hydroxyshogaol, 6-, 8-, 10-dehydroshogaol, diarylheptanoids
Inhibitory effects on the proliferation of human colon cancer cells
(i) Arrest at G0/G1 phase(ii) Reduced DNA synthesis
[103]
Grifola frondosa
Fruit
HT-29
10 ng/mL
Aqueous
Phenolic compounds (pyrogallol, caffeic acid, myricetin, protocatechuic acid)
Inhibition of TNBS-induced rat colitis
Induced cell cycle progression in G0/G1 phase
[104]
Cucumaria frondosa
The enzymatically hydrolyzed epithelium of the edible
HCT116
<150μg/mL
Hydroalcoholic
Monosulphated triterpenoid glycoside frondoside A, the disulphated glycoside frondoside B, the trisulphated glycoside frondoside C
Inhibition of human colon cancer cell growth
(i) Inhibition at S and G2-M phases with a decrease in Cdc25c and increase in p21WAF1/CIP(ii) Apoptosis associated with H2AX phosphorylation and caspase 2
[105]
Rolandra fruticosa
Leaf & twigs
HT-29
10 and 5 mg/kg/day
Methanolic
Sesquiterpene lactone (13-acetoxyrolandrolide)
Antiproliferative effect against human colon cancer cells
Inhibition of the NFκB pathway, NFκB subunit p65 (RelA), upstream mediators IKKβ and oncogenic K-ras
[106]
Cydonia oblonga Miller
Leaf & Fruit
Caco-2
250–500μg/mL
Methanolic
Phenolic compound (flavonol and flavone heterosides, 5-O-caffeoylquinic acid)
Antiproliferative effect against human kidney and colon cancer cells
(i) Suppression of factor activation, nuclear factor-kB (NFκB) activation, protein-1 (AP-1) transcription factor, mitogen protein kinases (MAPKs), protein kinases (PKs), namely, PKC, growth-factor receptor- (GFR-) mediated pathways and angiogenesis(ii) Cell cycle arrest and induction of apoptosis, antioxidant, and anti-inflammatory effects
[107]
Morchella esculenta
Fruits
HT-29
820 mg/mL
Methylene chloride
Steroids (mainly ergosterol derivatives) & polysaccharides & galactomannan
Antioxidant activity in HT-29 colon cancer cells
Inhibition of NF-B activation in the NF-B assay
[108]
Sedum kamtschaticum
Aerial part
HT-29
0–0.5 mg/mL
Methanolic
Buddlejasaponin IV
Induced apoptosis in HT-29 human colon cancer cells
Induction of apoptosis via mitochondrial pathway by downregulation of Bcl-2 protein levels, caspase 3 activation, and subsequent PARP cleavage
[109]
Ginseng and Glycyrrhiza glabra
Leaf
HT-29
500μL
Aqueous
Uracil, adenine, adenosine, Li-glycyrrhetinic acid, quiritin
NM
Antiproliferative effect determination of the protein levels of p21, cyclin D1, PCNA, and cdk-2, which are the key regulators for cell cycle progression
[110]
Orostachys japonicus
Leaf & stem
HT-29
2 mg/mL
Aqueous
Flavonoids, triterpenoids, 4-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, polysaccharide
Antiproliferation in HT-29 colon cancer cells
Inhibited proliferation at G2 point of the cell cycle and apoptosis via tumor suppressor protein p53
[111]
Ginkgo biloba
Fruit & leaf
HT-29
20–320mg/L
Aqueous
Terpene lactones and flavonoid glycosides
(i) Inhibited progression of human colon cancer cells(ii) Induced HT-29 cell apoptosis
Increase in caspase 3 activities and elevation in p53 MRN reduction in Bcl-2 mRNA
[112]
Oryza sativa
Seed
HT-29, SW 480, HCEC
100μg/mL
Ethyl acetate
Phenolic compound (tricin, ferulic acid, caffeic acid, and methoxycinnamic acid)
Inhibition of the human colon cancer cell growth
(i) Induced apoptosis by enhanced activation of caspases 8 and 3(ii) Decrease of the number of viable SW480 and HCEC cells(iii) Reduced colony-forming ability of these cells
[113]
Cnidium officinale Makino
Root
HT-29
305.024/mL
Ethanolic
Osthole, auraptenol, imperatorin
Inhibited proliferation of human colon cancer cells (HT-29)
Inhibition of the cellular proliferation via G0/G1 phase arrest of the cell cycle and induced apoptosis
[114]
Cnidium officinale Makino
Root
HT-29
0.1-5 mg/mL
Aqueous
N-(3-(Aminomethyl)benzyl)acetamidine
Inhibited the invasiveness of cytokine-treated HT-29 cells through the Matrigel-coated membrane in a concentration-dependent manner
(i) Reduction of HT-29 cell invasion through the Matrigel(ii) Inhibited cytokine-mediated NO production, iNOS expression, and invasiveness of HT-29 cells(iii) Inhibited MMP-2 activity
[115]
Long pepper (PLX)
Fruit
HT-29 and HCT116
0.10 mg/mL
Ethanolic
Piperidine alkaloids, piperamides, piperlongumine
(i) Induction of apoptosis, following DNA fragmentation in HT-29 colon cancer cells in a time-dependent manner(ii) Induced caspase-independent apoptosis
Induced whole cell ROS production
[116]
Achyranthes aspera
Root
COLO 205
50-100 and 150-200μg/mL
Ethanolic (EAA) and aqueous (AAA) root extractsAqueous
Phenolic compounds
(i) Enhanced growth inhibitory effects of AAA towards COLO 205 cells in contrast to EAA(ii) Stimulatory role of AAA in the activation of cell cycle inhibitors
(i) Triggered mitochondrial apoptosis pathway and S phase cell cycle arrest(ii) Increased levels of caspase 9, caspase 3, and caspase 3/7 activity
[117]
Thymus vulgaris
Leaf
HCT116
0.2, 0.4, 0.6, 0.8 mg/mL
Carvacrol and thymol
Inhibited proliferation, adhesion, migration, and invasion of cancer cells
[118]
Dictyopteris undulata
NM
SW480
40μg/mL
Ethanolic
Cyclozonarone benzoquinone
NM
Induced apoptosis by reducing Bcl-2 levels, upregulating Bax, and disrupting the mitochondrial membrane potential, leading to the activation of caspases 3 and 9
[119]
Dendrobium microspermae
NM
HCT116
0.25, 0.5, 1.0 mg/mL
Methanolic
NM
NM
Upregulation of Bax and caspases 9 and 3 and downregulation of Bcl-2 expression of genes
[120]
Cannabis sativa
Dry flower & leaf
DLD-1 and HCT116
0.3–5μM
Methanolic
Cannabidiol, phytocannabinoids
Reduced cell proliferation in a CB1-sensitive
(i) Reduced AOM-induced preneoplastic lesions and polyps(ii) Inhibited colorectal cancer cell proliferation via CB1 and CB2 receptor activation
[121]
Phoenix dactylifera L.
Fruit
Caco-2
0.2 mg/mL
Aqueous
Phenolic acids (gallic, protocatechuic, hydroxybenzoic, vanillic, isovanillic, syringic, caffeic, ferulic, sinapic, p-coumaric, isoferulic), flavonoid glycosides (quercetin, luteolin, apigenin, and kaempferol), and anthocyanidins
Increasing beneficial bacterial growth and inhibition of proliferation of colon cancer cells
NM
[122]
Melia toosendan
Fruit
SW480, CT26
0, 10, 20, 30, 40, 50μg/mL
Ethanolic
Triterpenoids, flavonoids, polysaccharide, limonoids
NM
(i) Inhibited cell proliferation of SW480 and CT26 by promoting apoptosis as indicated by nuclear chromatin condensation and DNA fragmentation(ii) Induced caspase 9 activity which further activated caspase 3 and poly(ADP-ribose) polymerase cleavage, leading the tumor cells to apoptosis
[123]
Crocus sativus L.
Flower
HCT116
0.25, 0.5, 1, 2, 4μg/mL
Ethanolic
Carotenoid, pigment, crocin, crocetin
Induced DNA damage and apoptosis
(i) Induction of a p53 pattern-dependent caspase 3 activation with a full G2/M stop(ii) Induced remarkable delay in S/G2 phase transit with entry into mitosis
[124]
Tepals and leaf
Caco-2
0.42 mg/mL
NM
Polyphenols, glycosides of kaempferol, luteolin, and quercetin
Proliferation of Caco-2 cells was greatly inhibited
NM
[125]
Luffa echinata
Fruit
HT-29
50, 100, and 200μg/mL
Methanolic
Amariin, echinatin, saponins, hentriacontane, gypsogenin, cucurbitacin B, datiscacin, 2-O-β-D-glucopyranosyl cucurbitacin B, and 2-O-β-D-glucopyranosyl cucurbitacin S
Increase in the population of apoptotic cells
(i) Inhibited the cellular proliferation of HT-29 cells via G2/M phase arrest of the cell cycle(ii) Induced apoptotic cell death via ROS generation(iii) Accumulation of caspase 3 transcripts of HT-29 cells
[126]
Vitis aestivalis hybrid
Fruits (wine)
CCD-18Co
25, 50, 100μg/mL
NM
Polyphenolics
NM
(i) Decreased mRNA expression of lipopolysaccharide- (LPS-) induced inflammatory mediators NFκB, ICAM-1, VCAM-1, and PECAM-1(ii) Enhanced expression of miR-126(iii) Decreased gene expression and reduced activation of the NFκB transcription factor, NFκB-dependent(iv) Decrease in ROS 113MAH
[127]
Xylopia aethiopica
Dried fruit
HCT116
0, 5, 10, 15, 20, 25, 30μg/mL
Ethanolic
Ent-15-oxokaur-16-en-19-oic acid (EOKA)
NM
(i) Induced DNA damage, cell cycle arrest in G1 phase, and apoptotic cell death
[128]
Sorghum
Grain
ER-β; nonmalignant young adult mouse colonocytes
1, 5, 10, 100μg/mL
Aqueous
Flavones (luteolin and apigenin), 3-deoxyanthocyanins naringenin (eriodictyol and naringenin)
Reduced cell growth via apoptosis
Increased caspase 3 activity
[129]
NM
HT-29, HCT116
0.9-2.0 mg/mL
Hydroethanolic
Procyanidin B1, delphinidin-3-O-glucoside, tannin, cyanidin-3-O glucoside
(i) Significantly arrested HT-29 cells in G1(ii) Highest growth inhibition(iii) Increased percentage of apoptotic cells
(i) Downregulation of apoptotic proteins, such as cIAP-2, livin, survivin, and XIAP, was seen in HCT116 cells(ii) Inhibition of tyrosine kinase
[130]
Panax notoginseng (Burk.) F.H. Chen
Root
LoVo and Caco-2
0, 100, 250, and 500μg/mL
Alcoholic
Saponin, ginsenoside
NM
Delay in progression of the G0/G1, S, or G2/M cell cycle phases
[131]
Brassica oleracea L. var. italica
Broccoli florets
HCT116
0, 1, 2.5, 5, 10μg/mL
Ethanolic
Glucoiberin, 3 hydroxy,4(α-L-rhamnopyranosyloxy), benzyl glucosinolate 4-vinyl-3-pyrazolidinone 4-(methyl sulphinyl), butyl thiourea, β-thioglucoside N-hydroxysulphates
NM
NM
[132]
Cistanche deserticola
Dried stem
SW480
In vivo: 0.4 g/kg/dayIn vitro: 100 μg/mL
Aqueous
Polysaccharides, phenylethanoid glycosides
(i) Decreased number of mucosal hyperplasia and intestinal helicobacter infection(ii) Increased number of splenic macrophage, NK cells, and splenic macrophages
Decreased frequency of hyperplasia andHelicobacter hepaticus infection of the intestine
[133]
Chaenomeles japonica
Fruit
Caco-2 and HT-29
10, 25, 50, 75, 100, 125, 150μM CE
NM
Procyanidins
NM
NM
[134]
Prunus mume
Fruit
SW480, COLO, and WiDr
150, 300, and 600μg/mL
Hydrophobic
Triterpenoid saponins
NM
(i) Inhibited growth and lysed SW480, COLO, and WiDr(ii) Induction of massive cytoplasmic vacuoles
[135]
Solanum lyratum
NM
COLO 205
50, 100, 200, 300, 400μg/mL
EtOH
β-Lycotetraosyl
Induced S phase arrest and apoptosis
(i) Induced DNA fragments(ii) Increased the levels of p27, p53, cyclin B1, active-caspase 3, and Bax(iii) Decreased the levels of Cdk1, pro-caspase 9, Bcl-2 and NF-ÎB, p65, and p50
[136]
Onopordum cynarocephalum
Aerial parts
HCT116, HT-29
0, 0.04, 0.12, 0.2, 0.4, 1.2 mg/mL0, 0.2, 0.4, 1.2, 2.0, 3.0 mg/mL
Aqueous
Flavonoids, lignans, and sesquiterpene lactones
NM
(i) Increase in the expression of proapoptotic proteins such as p53, p21, and Bax(ii) Inhibition of the antiapoptotic protein Bcl-2(iii) Decrease in cyclin D1 protein
[137]
Eleutherine palmifolia
Bulbs
SW480
2.5, 5, 10μg/mL
MeOH
Eleutherin, isoeleutherin
NM
(i) Inhibited the transcription of TCF/β-catenin(ii) Decrease in the level of nuclear β-catenin protein
[138]
Asparagus officinalis
Spears
HCT116
76μg/mL
Acetone
Steroidal saponins (HTSA-1, HTSAP-2, HTSAP-12, HTSAP-6, HTSAP-8)
NM
(i) Inhibition of Akt, p70S6K, and ERK phosphorylation(ii) Induction of caspase 3 activity, PARP-1 cleavage, DNA fragmentation, G0/G1 cell cycle arrest by reducing the expression of cyclins D, A, and E
[139]
Phyllanthus emblica L.
Seed, pulp
HCCSCs, HCT116
200μg/mL
Methanolic
Trigonelline, naringin, kaempferol, embinin, catechin, isorhamnetin, quercetin
(i) Suppressed proliferation(ii) Induced apoptosis independent from p53 stemness property (in HCCSCs)(iii) Antiproliferative properties
(i) Suppressed cell proliferation and expression of c-Myc and cyclin D1(ii) Induced intrinsic mitochondrial apoptotic signaling pathway
[140]
Red grape
NM
HT-29, HCT116
0.9-2.0 mg/mL
Hydroethanolic
Delphinidin glycosides, quercetin derivatives, delphinidin-3-O-glucoside (high), cyanidin-3-O-glucoside
(i) Highest growth inhibition(ii) Increased the percentage of apoptotic cells
(i) Downregulation of apoptotic proteins, such as cIAP-2, livin, survivin, and XIAP(ii) Inhibition of tyrosine kinase
[130]
Black lentil
NM
HT-29, HCT116
0.9-2.0 mg/mL
Hydroethanolic
Delphinidin glycosides, procyanidin B1, delphinidin-3-O-glucoside (high), cyanidin-3-O-glucoside
(i) Significantly arrested HT-29 cells in G1(ii) Highest growth inhibition(iii) Increased percentage of apoptotic cells
(i) Downregulation of apoptotic proteins, such as cIAP-2, livin, survivin, and XIAP(ii) Inhibition of tyrosine kinase
[130]
Graptopetalum paraguayense
Leaf
Caco-2, BV-2
0.2, 0.4, 0.6, 0.8, 1.0 mg/mL
Hydroethanolic
Oxalic acid, hydroxybutanedioic acid, gallic acid, quercetin, chlorogenic acid glucans with fucose, xylose, ribose (GW100) arabino-rhamnogalactans (GW100E)
(i) Great potential in antiproliferation(ii) Significant immunomodulatory activities on BV-2 cells and interleukin-6 (IL-6) (GW100)
(i) Scavengingα, α-diphenyl-β-picrylhydrazyl radicals (DPPH) (GW100E excelled in scavenging DPPH), 2,2-azino-bis [3-ethylbenzothiazoline-6-sulfonic acid] radicals (ABTS), superoxide anions (O2) (GW100)(ii) Significant inhibition of tumor necrosis factor-a (TNF-a), scavenging ABTS and O2
[141]
Butea monosperma
Flower
SW480
200, 370μg/mL
Floral
n-Butanol
Significant antiproliferative effect
(i) Significantly downregulated the expression of Wnt signaling proteins such asβ-catenin, APC, GSK-3β, cyclin D1, and c-Myc(ii) Increased intracellular level of ROS
[142]
Rehmannia glutinosa
NM
CT26
5, 20, 80μM
NM
Catalpol
Inhibited proliferation and growth invasion of colon cancer cells
(i) Downregulated MMP-2 and MMP-9 protein expressions(ii) Reduction in the angiogenic markers secretions
[143]
Telectadium dongnaiense
Bark
HCT116
1.5, 2.0μg/mL
MeOH extract
4-Dicaffeoylquinic acid, quercetin 3-rutinoside, periplocin
NM
(i) Inhibition ofβ-catenin/TCF transcriptional activity and effects on Wnt/β-catenin(ii) Downregulation of the expression of Wnt target genes
[144]
Gloriosa superba
Root
SW620
30 ng/mL
Protein hydrolysate extract
Protein hydrolysate
NM
(i) Upregulation of p53(ii) Downregulation of NFκB
[145]
Boswellia serrata
Resin
HT-29
100, 150μg
Methanolic
Boswellic acid
Decreased cell viability
(i) Reduction in mPGES-1, VEGF, CXCR4, MMP-2, MMP-9, HIF-1, PGE2 expression(ii) Increment in the caspase 3 activity(iii) Inhibition of cell migration and vascular sprout formation
[146]
Typhonium flagelliforme
Leaf
WiDr
70μg/mL
Ethyl acetate
Glycoside flavonoid, isovitexin, alkaloids
NM
Inhibition of COX-2 expression
[28]
Diospyros kaki
Fruit
HT-29
2,000μg/mL
Hydroacetone extract
Polyphenol
Impaired cell proliferation and invasion
NM
[147]
Carpobrotus edulis
Leaf
HCT116
1,000 mg/mL
Hydroethanolic
Gallic acid, quercetin, sinapic acid, ferulic acid, luteolin 7-o-glucoside, hyperoside, isoquercitrin, ellagic acid, isorhamnetin 3-O-rutinoside
Inhibited proliferation
(i) Possession of high DPPH scavenging activity and effective capacity for iron binding(ii) Inhibition of NO radical, linoleic acid peroxidation, protein glycation, and oxidative damage
[148]
Piper methysticum
Root
HT-29
10, 20, 30, 40, 50μg/mL
Aqueous
11-Hydroxy-12-methoxydihydrokavain, 11-hydroxy-12-methoxydihydrokavain, prenyl caffeate, pinostrobin chalcone, 11-methoxytetrahydroyangonin, awaine, methysticin, dihydromethysticin, 5,6,7,8-tetrahydroyangonin, kavain, 7,8-dihydrokavain, yangonin, desmethoxyyangonin, flavokawain B
Inhibited the growth
NM
[26]
Salvia ballotiflora
Ground aerial parts
CT26
6.76μg/mL
Hexane-washed chloroform extract
19-Deoxyicetexone, 7,20-dihydroanastomosine, icetexone, 19-deoxyisoicetexone
Cytotoxic activity
NM
[149]
Tinospora cordifolia
Stem
HCT116
1, 10, 30, 50μM
Hydroalcoholic
Clerodane furano diterpene glycoside, cordifoliosides A and Β, sitosterol, ecdysterone, 2β,3β:15,16-diepoxy-4α, 6β-dihydroxy-13(16),14-clerodadiene-17,12:18,1-diolide
Induced chromatin condensation and fragmentation of nuclei of few cells
(i) Considerable loss of MMP(ii) Decreased in mitochondria function(iii) Increased cytochrome c in the cytosol(iv) Induced ROS/oxidative stress(v) Increased autophagy
[150]
Euterpe oleracea
Fruit
NM
35μg/mL
Hydroethanolic
Vanillic acid, orientin, isoorientin
NM
(i) Scavenging capacity towards ROO and HOCl(ii) Inhibition of nitroso compound formation
[151]
Salvia miltiorrhiza
NM
HCT116
7.4±1.0, 4.4±0.5μg/mL
Ethanolic
Diterpene quinone
NM
Decreased levels of pro-caspases 3 and 9
[152]
Coffea
Bean
HCT116
1 mg/mL
Aqueous
Chlorogenic acid complex (CGA7)
NM
(i) DNA fragmentation, PARP-1 cleavage, caspase 9 activation, downregulation of Bcl-2 and upregulation of Bax
[153]
Illicium verum
Fruit
HCT116
10 mg/mL
Ethanolic
Gallic acid quercetin
Induction of apoptosis and inhibition of key steps of metastasis
NM
[154]
Garcinia propinqua Craib
Leaf
HCT116
NM
CH2Cl2 extract
Benzophenones, xanthones, and caged xanthones
Potent inhibitory cytotoxicities
NM
[155]
Stem, bark
HCT116
14.23, 23.95μM
MeOH, CH2Cl2, and EtOAc extract
Xerophenone A, doitunggarcinones A and B, sampsonione, 7β-H-11-benzoyl-5α-ydroxy-6, 10-tetramethyl-1-(3-methyl-2-butenyl)-tetracyclotetradecane-2,12,14-trione, hypersampsone M, assiguxanthone A (cudraxanthone Q), 40 10-O-methylmacluraxanthone (16), 41- and 5-O-methylxanthone V1
NM
NM
[156]
Malus pumila Miller cv. Annurca
Fruit
Caco-2
400 mg/L
Methanolic
Chlorogenic acid, (+)catechin, (–)epicatechin, isoquercetin, rutin, phloridzin, procyanidin B2, phloretin, quercetin
WNT inhibitors and reduced WNT activity elicited by WNT5A
NM
[157]
Malus domestica cv. Limoncella
Fruit
Caco-2
400 mg/L
Methanolic
Chlorogenic acid, (+)catechin, (–)epicatechin, isoquercetin, rutin, phloridzin, procyanidin B2, phloretin, quercetin
WNT inhibitors and reduced WNT activity elicited by WNT5A
NM
[157]
Coix lacryma-jobi var. ma-yuen
Leaf
HCT116
0.5, 1 mg/mL
Aqueous
Coixspirolactam A, coixspirolactam B, coixspirolactam C, coixlactam, methyl dioxindole-3-acetate
NM
Inhibited migration, invasion, and adhesion via repression of the ERK1/2 and Akt pathways under hypoxic conditions
[158]
Mesua ferrea
Stem, bark
HCT116, HT-29
3.3, 6.6, and 11.8μg/mL
NM
Fractions (α-amyrin, SF-3, n-Hex)
Downregulation of multiple tumor promoter
Upregulation of p53, Myc/Max, and TGF-β signaling pathways
[159]
Taraxacum
Root
SGC7901, BGC823
3 mg/mL
Aqueous
NM
NM
Proliferation and migration through targeting lncRNA-CCAT1
[160]
Portulaca oleracea
Leaf
HT-29 CSCs
2.25μg/mL
Alcoholic
Oxalic, malic acid
NM
Inhibited expression of the Notch1 andβ-catenin genes, regulatory and target genes that mediate the Notch signal transduction pathway
[161]
Hordeum vulgare L.
NM
HT-29
NM
Aqueous & juice
Protein, dietary fiber, the B vitamins, niacin, vitamin B6, manganese, phosphorus, carbohydrates
(i) Inhibited proliferation of cancer cells(ii) Cytotoxic activity
Free radical scavenging activity
[162]
Paraconiothyrium sp.
NM
COLO 205 and KM12
12.5μM
Methyl ethyl ketone extract
n-Hexane, CH2Cl2, EtOAc, and MeOH fractions (A−D)
(i) Growth inhibitory activity(ii) Antiproliferative effect
NM
[163]
Mentha×piperita
Leaf
HCT116
5, 10, 20, 30, 40, 50μg/mL
Aqueous
Polyphenols
NM
Inhibited replication of DNA and transcription of RNA which induce the ROS
[164]
Mammea longifolia Planch. and Triana
Fruit
SW480
25, 50, 100μg/mL
Methanolic
NM
NM
Mitochondria-related apoptosis and activation of p53
[165]
Rollinia mucosa (Jacq.) Baill.
NM
HCT116, SW-480
<4, <20μg/mL
EtOH
Rollitacin, jimenezin, membranacin, desacetyluvaricin, laherradurin
Cytotoxic activity
NM
[54]
Annona diversifolia Saff.
NM
SW-480
0.5μg/mL
NM
Cherimolin-2
Cytotoxic activity
NM
[54]
A. purpurea Moc. & Sessé ex Dunal
NM
HT-29
1.47μg/mL
CHCl3-MeOH
Purpurediolin, purpurenin, annoglaucin, annonacin A
Cytotoxic activity
NM
[54]
Viguiera decurrens (A.Gray) A. Gray
NM
NM
3.6μg/mL
Hex; EtOAc; MeOH
β-Sitosterol-3-O-β-D-glucopyranoside; β-D-glucopyranosyl oleanolate; β-sitosterol-3-O-β-D-glucopyranoside, and oleanolic acid-3-O-methyl-β-D-glucuronopyranoside ronoate
Cytotoxic activity
NM
[54]
Helianthella quinquenervis (Hook.) A. Gray
NM
HT-29
2-10μg/mL
NM
Demethylencecalin
Cytotoxic activity
NM
[54]
Smallanthus maculatus (Cav.) H. Rob.
NM
HCT15
<20μg/mL
Acetone
Fraction F-4, fraction F-5, ursolic acid
Cytotoxic activity
NM
[54]
Bursera fagaroides (Kunth) Engl.
NM
HF6
1.8×10-4 to 2.80 μg/mL
Hydroalcoholic
Podophyllotoxin,β-peltatin-A methyl ether, 5′-desmethoxy-β-peltatin-A methyl ether, desmethoxy-yatein, deoxypodophyllotoxin, burseranin, acetyl podophyllotoxin
NM
(i) Inhibitor of microtubules(ii) Ability to arrest cell cycle in metaphase
[54]
Viburnum jucundum C.V. Morton
NM
HCT15
<20μg/mL
Acetone
Ursolic acid
Cytotoxic activity
NM
[54]
Hemiangium excelsum (Kunth) A.C.Sm.
NM
HCT15
<10 (μg/mL)
MeOH
PE, EtOAc, MeOH
Cytotoxic activity
NM
[54]
Hyptis pectinata (L.) Poit.
NM
Col2
<4, <20μg/mL
NM
Pectinolide A, pectinolide B, pectinolide C,α-pyrone, boronolide, deacetylepiol-guine
Cytotoxic activity
NM
[54]
H. verticillata Jacq.
NM
Col2
<4,<20μg/mL
NM
Dehydro-β-peltatin, methyl ether dibenzylbutyrolactone, (-)-yatein, 4′-demethyl-deoxypodophyllotoxin
Nonspecific cytotoxic activity
NM
[54]
H. suaveolens (L.)
NM
HF6
2.8-12μg/mL
Chloroform and butanol
β-Apopicropodophyllin
Nonspecific cytotoxic activity
NM
[54]
Salvia leucantha Cav.
Leaf, root, stem
HF6, HT-29, HCT15
14.9, 12.7, 9.9μg/mL
CHCl3
NM
Cytotoxic activity
NM
[54]
Vitex trifolia L.
NM
HCT15
3.5 to <1 (μg/mL)
Hexane and dichloromethane
Salvileucalin B, Hex: leaf, Hex: stem, DCM: leaf, DCM: stem
Cytotoxic activity
NM
[54]
Persea americana Mill.
NM
HT-29
<4μg/mL and <20 μg/mL
Ethanolic
1,2,4-trihydroxynonadecan, 1,2,4-trihydroxyheptadec-16-ene, 1,2,4-trihydroxyheptadec-16-yne
Cytotoxic activity
NM
[54]
Linum scabrellum
Roots, aerial parts
HF6
0.2, 0.5, 2.3μg/mL
Chloroform and butanol
DCM: MeOH, 6MPTOXPTOX
NM
(i) Induction of cell cycle arrest in G2/M(ii) Inhibition of tubulin polymerization
[54]
Phoradendron reichenbachianum (Seem.) Oliv.
NM
HCT15
3.6, 3.9, and 4.3μg/mL
NM
Moronic acid
Cytotoxic activity
NM
[54]
Cuphea aequipetala Cav.
NM
HCT15
18.70μg/mL
Acetone
NM
Cytotoxic inactivity
NM
[54]
Galphimia glauca Cav.
NM
HCT15
0.63, 0.50, 1.99μg/mL
EtOH, MeOH, aqueous
NM
Cytotoxic activity
NM
[54]
Mimulus glabratus Kunth
NM
HF6
12.64μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
Picramnia antidesma Sw.
NM
HCT15
0.6 to 4.5μM
NM
10-Epi-uveoside, uveoside, picramnioside E, picramnioside D
Cytotoxic activity
NM
[54]
Penstemon barbatus (Cav.) Roth
NM
HF6
15.19μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
P. campanulatus (Cav.) Willd.
NM
HF6
6.74μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
Veronica americana Schwein. ex Benth.
NM
HF6
0.169 and 1.46μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
Zea mays L.
NM
HCT116, SW-480, SW-620
NM
NM
13-Hydroxy-10-oxo-trans-11-octadecenoic acid
Cytotoxic activity
NM
[54]
Colubrina macrocarpa (Cav.) G. Don
NM
HCT15
10, 2.1, 9.1μg/mL
PE, EtOAc, MeOH
NM
Cytotoxic activity
NM
[54]
Coix lacryma-jobi
Seed, endosperm, and hull
HT-29
0.1–1,000μg/mL
Methanolic, hexane
Phytosterols (campesterol, stigmasterol, andβ-sitosterol), gamma-linolenic acid (GLA), arachidonic acid (AA), eicosapentaenoicacid (EPA) and docosahexaenoic acid (DHA), linoleic acid
NM
(i) Influence of signal transduction pathways that involve the membrane phospholipids(ii) Enhancement of ROS generation and decrease of cell antioxidant capacity
[166]
Abutilon indicum
Leaf
HT-29
210μg/mL
Aqueous
Flavonoids (4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl, 2-ethoxy-4-vinylphenol, N,N′-dimethylglycine, lup-20(29)-en-3-one, linolenin, 1-mono-, 9-hexadecanoic acid methyl ester, linolenic acid methyl ester), phenolic (amino acids, terpenoids, fatty acids, methyl palmitoleate)
NM
(i) Increase in the levels of reactive oxygen species and simultaneous reduction in cellular antioxidant, mitochondrial membrane loss, DNA damage, and G1/S phase cell cycle arrest
[167]
Galla rhois
NM
HCT116, HT-29
12.5, 25, 50, 100, 200μg/mL
Aqueous with steaming process
Gallotannins
Increased contents of gallic acid and ellagic acid
(i) Induced apoptosis through the activation of caspases 3, 8, 9(ii) Modulated activation of mitogen and protein kinases, p38, and c-Jun NH2-terminal kinase
[168]
Artemisia annua Linné
Powder
HCT116
20, 30, 40, 60, 80, 100μg/mL
Ethanolic
Phenolic compounds
Inhibited cell viability and increased LDH release
(i) PTEN/p53/PDK1/Akt signal pathways through PTEN/p53 induce apoptosis(ii) Increased apoptotic bodies, caspase 3 and 7 activation(iii) Regulated cytochrome c translocation to the cytoplasm and Bax translocation to the mitochondrial membrane
[169]
Nelumbo nucifera stamen
Powder
HCT116
100, 200, 400μg/mL
Ethanolic crude
NM
NM
(i) Increased the sub-G1 population, mRNA levels of caspases 3 and 8, levels of IκBα and caspase 9(ii) Modulated the Bcl-2 family mRNA expression(iii) Reduced the mRNA levels of NFκB
[170]
Corn silk
NM
LoVo, HCT116
1.25, 2.5, 5, 10, 20μg/mL
Aqueous
Proteins, polysaccharides, flavonoid, vitamins, tannins, alkaloids, mineral salts, steroids
NM
(i) Increase in the Bax, cytochrome c, caspases 3 and 9 levels
[171]
Lycium barbarum L.
Powder
HT-29
1, 2, 3, 4, 5μg/mL
NM
Neoxanthin, all-trans-β-cryptoxanthin, polysaccharides, carotenoids, flavonoids
NM
(i) Upregulation of p53 and p21 expression(ii) Downregulation of the CDK2, CDK1, cyclin A, and cyclin B expression(iii) Arrest in the G2/M phase of cell cycle
[172]
Chrysobalanus icaco L.
Freeze-dried fruit
HT-29
1, 2.5, 5, 10, 20μg/mL
Crude ethyl acetate
Delphinidin, cyanidin, petunidin, and peonidin
NM
(i) Increased intracellular ROS production(ii) Decreased TNF-α, IL-1β, IL-6, and NFκB1 expressions
[173]
Zanthoxylum piperitum De Candolle
Fruit
Caco-2, DLD-1
200μg/mL
Aqueous
NM
NM
(i) Increased the phosphorylation of c-Jun N-terminal kinase (JNK)
[174]
Celtis aetnensis (Tornab.) Strobl (Ulmaceae)
Twigs
Caco-2
5, 50, 100, 250, or 500μg/mL
Methanolic
Flavonoid and triterpenic compounds
NM
(i) Increase in the levels of ROS(ii) Decrease in RSH levels and expression of HO-1
[175]
Rosa canina
Peel and pulp
Caco-2
62.5, 125, 250, 500μg/mL
Total extract (fraction 1), vitamin C (fraction 2), neutral polyphenols (fraction 3), and acidic polyphenols (fraction 4)
Polyphenols
Decreased production of reactive oxygen species (ROS)
NM
[176]
Rhazya stricta
Leaf
HCT116
47, 63, 79, and 95μg/cm2
Crude alkaloid
Alkaloids
NM
(i) Downregulated DNA-binding and transcriptional activities of NFκB and AP-1 proteins(ii) Increase in Bax, caspases 3/7 and 9, p53, p21 and Nrf-2 levels(iii) Decrease in expression of ERK MAPK, Bcl-2, cyclin D1, CDK-4, survivin, and VEGF
[177]
Green coffee
NM
Caco-2
10-1,000μg/mL
NM
5-Caffeoylquinic acid (5-CQA), 3,5-dicaffeoylquinic acid (3,5-DCQA), ferulic acid (FA), caffeic acid (CA), dihydrocaffeic acid (DHCA), dihydroferulic acid (DHFA)
Reduced viability of cancer cells
NM
[178]
Flourensia microphylla
Leaf
HT-29
NM
Ethanolic and acetone
Phenolic compounds
NM
(i) Inhibition of IL-8(ii) Activation of apoptosis by the increment of the Bax/Bcl-2 ratio and expression of TNF family
[179]
∗NM: not mentioned.


#### 3.1.2. Studies in Animal Models

The most used animal model is the murine one (Tables2(a) and 2(b)). In particular, studies were carried out above all on HT-29 and HCT116 cells. The effects of the different medicinal plants and their extracts are essentially the same detected in in vitro studies. In particular, plant extracts were able to induce apoptosis and inhibit proliferation and tumor angiogenesis by regulating p53 levels and checkpoint proteins with consequent cell cycle arrest and antiproliferative and antiapoptotic effects on cancerous cells.Table 2
(a)
Efficacy of medicinal plants on colon cancer inin vivo models
Scientific name
Parts used
Model
Dose
Type of extract
Important compounds
Cellular effect
Mechanisms
References
Vitis vinifera
Seed
In vivo (murine)
Caco-2
In vivo: 400–1,000 mg/kgIn vitro: 10–25 μg/mL
Aqueous
Procyanidins
(i) Increased crypt depth and growth-inhibitory effects(ii) Inhibited cell viability(iii) Significantly decreased the histological damage score
Reduced MPO (myeloperoxidase) activity
[180]
Seed
In vivo
HT-29, SW480
5 mg/kg
Aqueous
NM
NM
Decreased VEGF, TNF, MMP-1, MMP-3, MMP-7, MMP-8, MMP-9, and MMP-13 protein expression
[181]
Skin
In vivo
NM
7.5, 30, 60μg/mL
Methanolic
4′-Geranyloxyferulic acid
NM
NM
[30]
Seed
In vivo (murine)
NM
0.12%w/w
NM
Catechin, epicatechin
NM
(i) Suppressed proliferation, sphere formation, nuclear translocation ofβ-catenin and Wnt/β-catenin signaling(ii) Elevated p53, Bax/Bcl-2 ratio, and cleaved PARP and mitochondrial-mediated apoptosis
[31]
Camellia sinensis
Leaf
In vivo (murine)
HT-29
In vitro: 0, 10, 30, 50μMIn vivo: 1.5 mg per day
Aqueous
Catechin, epigallocatechin gallate
1.9-fold increase in tumor endothelial cell apoptosis
Inhibited the ERK-1 and ERK-2 activation, VEGF expression, and VEGF promoter
[182]
In vivo (murine)
HCT116
0.5%
NM
NM
Reduced basement membrane
Inhibition of MMP-9 and VEGF secretion
[183]
In vivo (murine)
Caco-2, HT-29
300μM
Aqueous
Theaflavins (TF-2, TF-3, TF-1)
Induced apoptosis of human colon cancer cells
Inhibition of edema formation correlated to attenuation of COX-2 expression and promoter analysis revealed modulation of NFκB, AP-1, CREB, and/or NF-Il-6 (C/EBP)
[36]
In vivo (murine)
HT115
25μg/mL
Hydroethanolic
Phenolic compounds (p-hydroxyphenyl ethanol, pinoresinol & dihydroxyphenyl ethanol)
NM
Inhibition via reduced expression of a range ofα5 & β1
[184]
Sasa quelpaertensis
Leaf
In vivo
HT-29, HCT116
0, 100, 200, 300 mg/L
Ethanolic
p-Coumaric acid, tricin
Inhibition of colony formation
(i) Nonadherent sphere formation suppressed CD133+ & CD44+ population(ii) Downregulated expression of cancer stem cell markers
[41]
Anoectochilus
NM
In vivo
CT26
Oral dose of 50 & 10 mg/mouse per day
Aqueous
Kinsenoside
Stimulated proliferation of lymphoid tissues
Activation of phagocytosis of peritoneal macrophages
[185]
Purple-fleshed potatoes
Fruit
In vivo
Colon cancer stem cells
5.0μg/mL
Ethanol, methanol, ethyl acetate
Anthocyanin,β-catenin, cytochrome c
Reduction in colon CSCs number and tumor incidence
(i) Increase in cytochrome c levels from p53 status and maybe mitochondria-mediated apoptosis(ii) Suppressed levels of cytoplasmic and nuclear β-catenin
[58]
Phaseolus vulgaris
Leaf
In vivo
HT-29
Nm
Ethanolic
Polysaccharides, oligosaccharides
Induction of apoptosis and inhibit proliferation
(i) Inactivation of the retinoblastoma phosphoprotein(ii) Induced G1 arrest(iii) Suppression of NF-jb1(iv) Increase in EGR1 expression
[59]
Rosmarinus officinalis L.
Leaf
In vivo
HT-29
SC-RE 30μg/mL and CA 12.5 μg/mL
Ethanolic
Polyphenols (carnosic acid (CA) and carnosol)
(i) Activation of Nrf2 transcription factor(ii) Activated common regulators, such as XBP1 (Xbp1) gene, SREBF1/SREBF2 (Srebp1/2), CEBPA and NR1I2 (Pxr) genes
Leaf
In vivo (rat)
NM
NM
Ethanolic
Rosmanol and its isomers, carnosol, rosmadial, carnosic acid, and 12-methoxycarnosic acid, carnosic acid, carnosol
Interactions with the gut microbiota and by a direct effect on colonocytes with respect to the onset of cancer or its progression
NM
Wasabia japonica
Rhizomes
In vivo
COLO 205
5 mg/mL
Methanolic
6-(Methylsulfinyl)hexyl isothiocyanate
Anticolon cancer properties through the induction of apoptosis and autophagy
(i) Activation of TNF-α, Fas-L, caspases(ii) Truncated Bid and cytochrome c(iii) Decreased phosphorylation of Akt and Mtor(iv) Promoted expression of microtubule-associated protein 1 light chain 3-II and AVO formation
[186]
Zingiberaceae
Rhizome
HT-29
HT-29
5 g/kg
Dichloromethanic
Turmerone
Suppressed the proliferation of HT-29 colon cancer cells
(i) LDH release(ii) ROS generation(iii) Collapse in mitochondrial membrane potential(iv) Cytochrome c leakage(v) Activation of caspase 9 and caspase 3
[187]
Panax quinquefolius
Root
In vivo (murine)
NM
30 mg/kg
Ethanolic
Ginsenosides (protopanaxadiol or protopanaxatriol)
Attenuated azoxymethane/DSS-induced colon carcinogenesis by reducing the colon tumor number and tumor load
(i) Reduced experimental colitis(ii) Attenuated on AOM/DSS-induced colon carcinogenesis(iii) Proinflammatory cytokines activation(iv) Suppressed DSS(v) Downregulated inflammatory cytokine gene expression
[188]
Myrtaceae
Leaf
In vivo (murine)
HCT116
100μg/mL (in vitro) 200 and 100 μg/disc (in vivo)
Methanolic
Phenolics, flavonoids, betulinic acid
Inhibition of tumor angiogenesis
(i) Inhibition of angiogenesis of tube formation on Matrigel matrix and HUVECS migration (in vitro)(ii) Decreased nutrient and oxygen supply and consequently tumor growth and tumor size (in vivo)(iii) Increased extent of tumor necrosis
[82]
Spica prunellae
Leaf
In vivo
HT-29
200 mg/mL (in vitro), 600 mg/mL (in vivo)
Ethanolic
Rosmarinic acid
Induction of apoptosis and inhibition of cell proliferation and tumor angiogenesis
(i) Induced apoptosis(ii) Inhibited cancer cell proliferation and angiogenesis STAT3 phosphorylation(iii) Regulated expression of Bcl-2, Bax, cyclin D1, CDK4, VEGF-A, and VEGFR-2 (in vivo)
[83]
Gymnaster koraiensis
Aerial part
In vivo (murine)
NM
500μmol/kg
Ethanolic
Gymnasterkoreaynes B, C, E, 2,9,16-heptadecatrien-4,6-dyne-8-ol
Anti-inflammatory and cancer preventive activities
(i) Significant decrease in expression of COX-2(ii) Increase in serum IL-6
[189]
Allium fistulosum
Edible portions
In vivo (murine)
CT26
50 mg/kg b.w.
Hot water
p-Coumaric acid, ferulic acid, sinapic acid, quercitrin, isoquercitrin, quercetol, kaempferol
Suppression of tumor growth and enhanced survival rate of test mice
(i) Decreased expression of inflammatory molecular markers(ii) Downregulated expression of MMP-9 and ICAM(iii) Metabolite profiling and candidate active phytochemical components
[190]
Annona squamosa Linn
Leaf
In vivo (animal)
HCT116
8.98μg/mL
Crude ethyl acetate
Acetogenins (annoreticuin & isoannoreticuin) and alkaloids dopamine, salsolinol, and coclaurine
(i) Inhibited growth and proliferation of tumor cells
Reactive oxygen species (ROS) formation, lactate dehydrogenase (LDH) release, and caspases 3/7, 8, 9 activation
[191]
Eupatorium cannabinum
Aerial parts
In vivo (murine)
HT-29
25μg/mL
Ethanolic
Pyrrolizidine alkaloids (senecionine, senkirkine, monocrotaline, echimidine)
Cytotoxicity against colon cancer cells
(i) Upregulation of p21 and downregulation of NCL, FOS, and AURKA, indicating reduced proliferation capacity(ii) Mitotic disruption and nonapoptotic cell death via upregulation of Bcl-xL
[96]
Flacourtia indica
Aerial parts
In vivo (murine)
HCT116
500μg/mL
Methanolic
Phenolic glucoside (flacourticin, 4′-benzoylpoliothrysoside)
Antiproliferative and proapoptotic effects in HCT116 cells
Apoptosis via generation of ROS and activation of caspases (PARP)
[192]
Sorghum bicolor
The dermal layer of stalk
In vivo (murine)
HCT116 & colon cancer stem cells
>16 and 103μg/mL
Phenolic, acetone
Apigeninidin & luteolinidin
Antiproliferative effect
(i) Target p53-dependent and p53-independent pathways
[97]
Gleditsia sinensis
Thorn
In vivo (murine)
HCT116
800μg/mL
Aqueous
Flavonoid, lupine acid, ellagic acid glycosides
Inhibited proliferation of colon cancer
(i) Increased p53 levels(ii) Downregulation of the checkpoint proteins, cyclin B1, Cdc2, and Cdc25c
[90]
Thorn
In vivo (murine)
HCT116
600μg/mL
Ethanolic
NM
Inhibitory effect on the proliferation of human colon cancer HCT116 cells
(i) Caused G2/M phase cell cycle arrest
[91]
Zingiber officinale
Rhizome
In vitro/in vivo (murine)
HCT116
5μM
Ethanolic
6-Paradol, 6- and 10-dehydrogingerdione, 6- and 10-gingerdione, 4-, 6-, 8-, and 10-gingerdiol, 6-methylgingerdiol, zingerone, 6-hydroxyshogaol, 6-, 8-, 10-dehydroshogaol, diarylheptanoids
Inhibitory effects on the proliferation of human colon cancer cells
(i) Arrest of G0/G1 phase(ii) Reduced DNA synthesis(iii) Induced apoptosis
[103]
Cucumaria frondosa
The enzymatically hydrolyzed epithelium of the edible
In vivo (murine)
HCT116
<150μg/mL
Hydroalcoholic
Monosulphated triterpenoid glycoside frondoside A, the disulphated glycoside frondoside B, the trisulphated glycoside frondoside C
(i) Inhibition at S and G2-M phase with a decrease in Cdc25c(ii) Increase in p21WAF1/CIP
(i) Inhibition the growth of human colon(ii) Apoptosis associated with H2AX phosphorylation and caspase 2
[105]
Rolandra fruticosa
Leaf & twigs
In vivo (murine)
HT-29
10 and 5 mg/kg/day
Methanolic
Sesquiterpene lactone (13-acetoxyrolandrolide)
Antiproliferative effect against human colon cancer cells
(i) Inhibition of the NFκB pathway, subunit p65 (RelA) and upstream mediators IKKβ and oncogenic K-ras
[106]
Cydonia oblonga Miller
Leaf & fruit
In vivo (murine)
Caco-2
250–500μg/mL
Methanolic
Phenolic compound (flavonol and flavone heterosides, 5-O-caffeoylquinic acid)
Antiproliferative effect against human kidney and colon cancer cells
(i) Suppression of NFκB activation, activator (AP-1), mitogen-activated protein kinases, namely, PKC, (GFR)-mediated pathways(ii) Cell cycle arrest(iii) Induction of apoptosis, antioxidant, and anti-inflammatory effects
[107]
Sedum kamtschaticum
Aerial part
In vivo (murine)
HT-29
0–0.5 mg/mL
Methanolic
Buddlejasaponin IV
Induced apoptosis in HT-29 human colon cancer cells
(i) Induced apoptosis via mitochondrial-dependent pathway triggered by downregulation of Bcl-2 protein levels, caspase 3 activation, and subsequent PARP cleavage
[109]
Ganoderma lucidum
Caps & stalks
In vivo (murine)
HT-29
0-0.1 mg/mL
Triterpene extract (hot water) extract
Polysaccharides (mainly glucans & glycoproteins), triterpenes (ganoderic acids, ganoderic alcohols, and their derivatives)
Cytokine expression inhibited during early inflammation in colorectal carcinoma
Induced autophagy through inhibition of p38 mitogen-activated kinase and activation of farnesyl protein transferase (FPT)
[193]
Ginkgo biloba
Fruit & leaf
In vivo (murine)
HT-29
20–320 mg/L
Aqueous
Terpene lactones and flavonoid glycosides
Inhibited progression of human colon cancer cells induced HT-29 cell apoptosis
(i) Activation in caspase 3, reduction in Bcl-2 expression, and elevation in p53 expression
[112]
Rubus occidentalis
Fruit
In vivo (murine)
JB6 Cl 41
25μg/mL
Methanolic
β-Carotene, α-carotene, ellagic acid, ferulic acid, coumaric acid
Inhibited tumor development
(i) Impaired signal transduction pathways leading to activation of AP-1 and NFB RU-ME fraction
[194]
Oryza sativa
Seed
In vivo (murine)
HT-29, SW 480, HCEC
100μg/mL
Ethyl acetate extract
Phenolic compound (tricin, ferulic acid, caffeic acid, and methoxycinnamic acid)
Inhibited growth of human colon cancer cells
(i) Induction of apoptosis by enhanced activation of caspases 8 and 3(ii) Decreased the number of viable SW480 and HCEC cells
[113]
Cistanche deserticola
Dried stem
In vivo (murine)
SW480
In vivo: 0.4 g/kg/dayIn vitro: 100 mg/mL
Aqueous
Polysaccharides, phenylethanoid glycosides
Decreased mucosal hyperplasia and helicobacter infection
(i) Increased number of splenic macrophages and NK cells(ii) Decreased frequency of hyperplasia and H. hepaticus infection of the intestine
[133]
Rehmannia glutinosa
NM
In vivo (male C57BL6 mice and Sprague-Dawley rats)
CT26
28 mg/kg
NM
Catalpol
(i) Inhibited proliferation, growth, and expression of angiogenic markers
(i) VEGF, VEGFR2, HIF-1α, bFGF inhibited the expressions of inflammatory factors such as IL-1β, IL-6, and IL-8
[143]
Olea europaea
Olive mill wastewater
In vivo (murine)
NM
NM
Methanolic
Hydroxytyrosol
Interferes with tumor cell growth
NM
[195]
Leaf
In vivo (xenograft model) (murine)
HCT116, HCT8
0, 5, 10, 20, 30, 50, and 70μg/mL
Phenolic
Oleuropein and hydroxytyrosol
NM
(i) Activation of caspases 3, 7, and 9(ii) Decrease of mitochondrial membrane potential and cytochrome c release(iii) Increase in intracellular Ca2+ concentration
[196]
Ginkgo biloba L.
Leaf
In vivo (rat)
NM
0.675 and 1.35 g/kg
Methanolic
Flavonoid glycosides, terpene lactones, and ginkgolic acids
(i) Suppressed tumor cell proliferation, promoted apoptosis, and mitigated inflammation
NM
[197]
Rhus trilobata Nutt.
NM
In vivo (hamster)
NM
400 mg/kg, 100 mg/kg
Aqueous
Tannic acid, gallic acid
Cytotoxic activity
NM
[54]
Annona diversifolia Saff.
NM
In vivo (mice)
SW-480
1.5, 7.5 mg/kg/day
NM
Laherradurin
Cytotoxic activity
NM
[54]
A. muricata L.
NM
In vivo (rat)
NM
250/500 mg/kg
EtOAc
A, B, and C, and cis- and trans-annomuricin-D-ones
Cytotoxic activity
NM
[54]
Plumeria acutifolia Poir.
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Lasianthaea podocephala (A. Gray) K. M. Becker
NM
In vivo (hamster)
NM
200 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Flourensia cernua DC.
NM
In vivo (hamster)
NM
350 mg/kg/day
Aqueous
Flavonoids, sesquiterpenoids, monoterpenoids, acetylenes, p-acetophenones, benzopyrans, benzofurans
Cytotoxic activity
NM
[54]
Ambrosia ambrosioides (Cav.) W. W. Payne
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Alnus jorullensis Kunth
NM
In vivo (hamster)
NM
175 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Dimorphocarpa wislizeni (Engelm.) Rollins
NM
In vivo (hamster)
NM
100 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Euphorbia pulcherrima Willd. ex Klotzsch
NM
In vivo (hamster)
NM
200 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Acalypha monostachya Cav.
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Crotalaria longirostrata Hook. & Arn.
NM
In vivo (hamster)
NM
400 mg/kg/day, 350 mg/kg/day
EtOH-CHCl3
NM
Cytotoxic activity
NM
[54]
Asterohyptis stellulata (Benth.) Epling
NM
In vivo (hamster)
NM
50 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Acacia constricta A. Gray
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Holodiscus dumosus A. Heller
NM
In vivo (hamster)
NM
350 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Butea monosperma
Flower
In vivo (rat)
HT-29
150 mg/kg
n-Butanol extract
Isocoreopsin, butrin, and isobutrin
Free radical scavenging and anticancer activities
NM
[198]
Taraxacum spp.
Root
In vivo (xenograft murine model)
HT-29, HCT116
40 mg/kg/day
Aqueous
α-Amyrin, β-amyrin, lupeol, and taraxasterol
Induced programmed cell death
NM
[199]
∗NM: not mentioned.
(b)
Other effects of medicinal plants inin vivo models
Scientific name
Parts used
Model
Dose
Type of extract
Important compounds
Cellular effect
Mechanisms
References
Allium sativum
Root
In vivo (murine)
NM
2.4 mL of daily
Ethanolic
Allicin, S-allylmercaptocysteine
Significantly suppressed both the size and number of colon adenomas
Enhancement of detoxifying enzymes: SAC and GST activity
[200]
Olea europaea
Fruit
In vivo
Caco-2
50μM
Aqueous
Phenolic compounds, authentic hydroxyl tyrosol (HT)
(i) Effect of OPE and HT on CB1 associated with reduced proliferation of Caco-2 cells(ii) Increase in CB1 expression in the colon of rats receiving dietary EVOO
Increase in Cnr1 gene expression, CB1 protein levels
[201]
In vivo (murine)
HT115
25μg/mL
Hydroethanolic
Phenolic compounds (p-hydroxyphenyl ethanol, pinoresinol & dihydroxyphenyl ethanol)
NM
Inhibition via reduced expression of a range ofα5 & β1
[184]
Origanum vulgare L.
Leaf
In vivo (murine)
NM
20, 40, 60 mg·kg−1
Aqueous
Rosmarinic acid, caffeic acid, flavonoids
Antioxidant status
(i) Increased LPO products and activity of SOD and CAT enzymes and GST and GPx activity(ii) Antioxidant and anticarcinogenic effect
[202]
Hazelnut
Skin
In vivo
NM
The flow rate 0.21 mL/min and injection volume 9.4μL
Aqueous
Flavan-3-ols, in monomeric and polymeric forms, and phenolic acids
(i) Decreased circulating levels of free fatty acids and triglycerides(ii) Higher excretion of bile acid
Increase of the total antioxidant capacity of plasma
[203]
Apples and apple juice
Fruit
In vivo
NM
90 mg/L
Aqueous
Phenolic acids, flavonoids, tannins, stilbenes, curcuminoids
NM
NM
[204]
Grifola frondosa
Fruit
In vivo (murine)
HT-29
10 ng/mL
Aqueous
Phenolic compounds (pyrogallol, caffeic acid, myricetin, protocatechuic acid, etc.)
Inhibition of TNBS-induced rat colitis
(i) Induced cell cycle progression in G0/G1 phase and apoptotic death
[104]
Ruta chalepensis
Leaf
In vivo (human)
NM
250μg/mL
Ethanolic
Rutin, gallic acid, catechin hydrate, naringin
Oxidative profile in patients with colon cancer
NM
[205]
Cannabis sativa
Dry flower & leaf
In vivo (murine)
DLD-1 and HCT116
0.3–5μM
Methanolic
Cannabidiol, phytocannabinoids
NM
(i) Reduced cell proliferation in a CB1-sensitive and AOM-induced preneoplastic lesions and polyps(ii) Inhibition of colorectal cancer cell proliferation via CB1and CB2 receptor activation
[121]
Melia toosendan
Fruit
In vivo (murine)
SW480, CT26
0, 10, 20, 30, 40, 50μg/mL
Ethanolic
Triterpenoids, flavonoids, polysaccharide, limonoids
NM
(i) Inhibited cell proliferation of SW480 and CT26 by promoting apoptosis as indicated by nuclear chromatin condensation and DNA fragmentation(ii) Induced caspase 9 activity which further activated caspase 3 and poly(ADP-ribose) polymerase cleavage, leading the tumor cells to apoptosis
[123]
Smallanthus sonchifolius
Root
In vivo (murine)
NM
73.90, 150.74, 147.65, and 123.26 mg/kg
Aqueous
Fructans
NM
Reduction incidence of colon tumors expressing alteredβ-catenin
[206]
Punica granatum
Peel
In vivo (adult male Wistar rats)
NM
4.5 g/kg
Methanolic
Gallic acid, protocatechuic acid, cateachin, rutin, ellagic acid, punicalagin
NM
(i) Reduction in TGF-β, Bcl-2, EGF, CEA, CCSA-4, MMP-7 and in COX-2, cyclin D1, survivin content(ii) Downregulated expression of β-catenin, K-ras, c-Myc genes
[207]
Linum usitatissimum
Seed
In vivo (male Sprague-Dawley rats)
NM
500 mg/kg
Alkaline
Secoisolariciresinol diglucoside, carbohydrates, proteins, and tannins
Reduced the serum fasting glucose levels
Significantly reduced the HbA1c, insulin levels, and proinflammatory cytokines
[208]
Diospyros kaki
Fruit
In vivo (male CD-1 mice)
NM
15 mg/kg
Hydroacetone
Polyphenol
(i) Decreased attenuation of colon length in diarrhea severity(ii) Reduced mortality rate(iii) Reduction of the extent of visible injury (ulcer formation) and of mucosal hemorrhage
Decreased expression of COX-2 and iNOS in the colonic tissue
[147]
Muntingia calabura
Leaf
In vivo (rat)
NM
50, 250, 500 mg/kg
Methanolic
Rutin, gallic acid, ferulic acid, and pinocembrin
Reduction of the colonic oxidative stress, increasing the antioxidants levels possibly via the synergistic action of several flavonoids
NM
[209]
Portulaca oleracea
NM
In vivo (murine)
HT-29 CSCs
2.25μg/mL
Alcoholic
NM
Regulatory and target genes that mediate the Notch signal transduction pathway
Inhibition of expression of the Notch1 andβ-catenin genes
[161]
Aloe vera
Gel
In vivo (murine)
NM
400 mg/kg/day
Gel
Polysaccharides
NM
(i) Via inhibition of the cell cycle progression(ii) Induction of cellular factors, such as extracellular signal-regulated kinases 1/2, cyclin-dependent kinase 4, and cyclin D1; on the other hand, PAG increased the expression of caudal-related homeobox transcription factor 2
[210]
Artemisia annua Linné
Powder
In vivo (xenograft murine model)
HCT116
20, 40 mg/kg/day
Ethanolic
Phenolic compounds
NM
(i) Induced apoptosis via PTEN/p53/PDK1/Akt signal pathways through PTEN/p53(ii) Inhibited cell viability and increased LDH release and apoptotic bodies, caspase 3 and 7 activation, and reduced mitochondria membrane potential(iii) Regulated cytochrome c translocation to the cytoplasm and Bax translocation to the mitochondrial membrane(iv) Regulation of proteins
[169]
Hordeum vulgare
Powder
In vivo (xenograft murine model)
HT-29
2 g/kg and 1 g/kg
Aqueous (fermented)
β-Glucan, protein, amino acids, phenolic compounds
NM
(i) Promoted tumor apoptosis by upregulating the mRNA expression of Bax and caspase 3 and downregulating the mRNA expression of Bcl-2 and cyclin D1(ii) Decreased mRNA expression of Bcl-2 and cyclin D1(iii) Upregulated expressions levels of Bax and caspase 3
[211]
Dendrophthoe pentandra
Leaf
In vivo (murine)
NM
125, 250, 500 mg/kg
Ethanolic
Quercetin-3-rhamnose
NM
(i) Decreased the levels of IL-22, MPO levels, proliferation of epithelial cells(ii) Inhibited S phase of the cell cycle(iii) Upregulated p53 wild-type gene expression
[212]
Aquilaria crassna
Stem, bark
In vivo (murine)
HCT116
2,000 mg/kg/day100, 200 mg/kg
NM
Resin and essential oils
NM
NM
[213]
Berberis integerrima
NM
In vivo (murine)
NM
50 and 100 mg/kg
Hydroalcoholic
NM
NM
NM
[214]
Salix aegyptiaca
Bark
In vivo (murine)
NM
100 and 400 mg/kg
Ethanolic
Catechin, catechol, and salicin
NM
Decreased level of EGFR, nuclearβ-catenin, and COX-2
[215]The main mechanisms of action of medicinal plants are summarized in Figure1.Figure 1
Cell damage and cancer trigger p53 activation. The p53 protein activates the apoptotic protein Bax. Bax inhibits the antiapoptotic protein Bcl-2. During apoptosis, cytochrome c is released from mitochondria. To activate the Apaf-1 protein, the interaction between these proteins and cytochrome C is necessary. Pro-caspase 9 attaches to Apaf-1 and activates caspase 9. Caspase 9 activates caspases 3 and 7 and apoptosis occurs.Inin vitro studies, it has been found that grapes, which contain substantial amounts of flavonoids and procyanidins, play a role in reducing the proliferation of cancer cells by increasing dihydroceramides and p53 and p21 (cell cycle gate keeper) protein levels. Additionally, grape extracts triggered antioxidant response by activating the transcriptional factor nuclear factor erythroid 2-related factor 2 (Nrf2) [27].Grape seeds contain polyphenolic and procyanidin compounds, and their reducing effects on the activity of myeloperoxidase have been shown inin vitro and in vivo studies. It has been suggested that grape seeds could inhibit the growth of colon cancer cells by altering the cell cycle, which would lead eventually to exert the caspase-dependent apoptosis [180].Another plant that attracted researchers’ attention was soybean, which contain saponins. After 72 h of exposure of colon cancer cells to the soy extract, it was found that this extract inhibited the activity and expression of protein kinase C and cyclooxygenase-2 (COX-2) [34]. The density of the cancer cells being exposed to the soy extract significantly decreased. Soybeans can also reduce the number of cancer cells and increase their mortality, which may be due to increased levels of Rab6 protein [216].Green tea leaves have also attracted the researchers’ attention in these studies. Green tea leaves, with high levels of catechins, increased apoptosis in colon cancer cells and reduced the expression of the vascular endothelial growth factor (VEGF) and its promoter activity inin vitro and in vivo studies. The extract increased apoptosis (programmed cell death) by 1.9 times in tumor cells and 3 times in endothelial cells compared to the control group [182]. In another in vitro study, the results showed that green tea leaves can be effective in the inhibition of matrix metalloproteinase 9 (MMP-9) and in inhibiting the secretion of VEGF [183].Garlic was another effective plant in this study. Its roots have allicin and organosulfur compounds. In anin vitro study, they inhibited cancer cell growth and induced apoptosis through the inhibition of the phosphoinositide 3-kinase/Akt pathway. They can also increase the expression of phosphatase and tensin homolog (PTEN) and reduce the expression of Akt and p-Akt [32]. Garlic roots contain S-allylcysteine and S-allylmercaptocysteine, which are known to exhibit anticancer properties. The results of a clinical trial on 51 patients, whose illness was diagnosed as colon cancer through colonoscopy, and who ranged in age from 40 to 79 years, suggest that the garlic extract has an inhibitory effect on the size and number of cancer cells. Possible mechanisms suggested for the anticancer effects of the garlic extract are both the increase of detoxifying enzyme soluble adenylyl cyclase (SAC) and an increased activity of glutathione S-transferase (GST). The results suggest that the garlic extract stimulates mouse spleen cells, causes the secretion of cytokines, such as interleukin-2 (IL2), tumor necrosis factor-α (TNF-α), and interferon-γ, and increases the activity of natural killer (NK) cells and phagocytic peritoneal macrophages [200].The results ofin vitro studies on olive fruit showed that it can increase peroxide anions in the mitochondria of HT-29 cancer cells due to the presence of 73.25% of maslinic acid and 25.75% of oleanolic acid. It also increases caspase 3-like activity up to 6 times and induces programmed cell death through the internal pathway [217]. Furthermore, the olive extract induces the production of reactive oxygen species (ROS) and causes a quick release of cytochrome c from mitochondria to cytosol.The pomegranate fruit contains numerous phytochemicals, such as punicalagins, ellagitannins, ellagic acid, and other flavonoids, including quercetin, kaempferol, and luteolin glycosides. The results of anin vitro study indicate the anticancer activity of this extract through reduction of phosphorylation of the p65 subunit and subsequent inhibition of nuclear factor-κB (NFκB). It also inhibits the activity of TNF receptor induced by Akt, which is needed for the activity of NFκB. The fruit juice can considerably inhibit the expression of TNF-α-inducing proteins (Tipα) in the COX-2 pathway in cancer cells [43]. The effective and important compounds in pomegranate identified in these 104 studies are flavonoids, polyphenol compounds, such as caffeic acid, catechins, saponins, polysaccharides, triterpenoids, alkaloids, glycosides, and phenols, such as quercetin and luteolin, and kaempferol and luteolin glycosides.In a systematic review of the plants being studied, some mechanisms were mainly common, including the induction of apoptosis by means of an increase of expression and levels of caspase 2, caspase 3, caspase 7, caspase 8, and caspase 9 in cancer cells, increasing the expression of the proapoptotic protein Bax and decreasing the expression of the antiapoptotic proteins.Many herbal extracts block specific phase of the cell cycle. For instance, the extract prepared from the leaves ofAnnona muricata inhibits the proliferation of colon cancer cells and induces apoptosis by arresting cells in the G1 phase [53]. They can also prevent the progress of the G1/S phase in cancer cells [74]. In general, the herbal extracts reported here have been able to stop cancer cells at various stages, such as G2/M, G1/S, S phase, G0/G1, and G1 phase, and could prevent their proliferation and growth.Other important anticancer mechanisms are the increase of both p53 protein levels and transcription of its gene. Even the increase of p21 expression is not without effect [137]. In an in vitro study on the Garcinia mangostana roots, the results were indicative of the inhibitory effect of the extract of this plant on p50 and P65 activation [93]. Moreover, reduction of cyclin D1 levels and increase of p21 levels are among these mechanisms [137], as well as inhibition of NFκB and reduction of the transcription of its genes, which contribute to reduce the number of cancerous cells [127]. Other important anticancer mechanisms are the inhibition of COX-2, as well as the reduction of the protein levels in this pathway [34]. In addition to this, in some cases, the inhibition of MMP-9 can be mentioned as the significant mechanism of some herbal extracts to kill cancer cells [183].


## 3.1. Medicinal Plants and Colon Cancer

Overall, 1,150 articles were collected in the first step and unrelated articles were excluded later on according to title and abstract evaluation. Moreover, articles that did not have complete data along with congress and conference proceedings were excluded. Accordingly, a total of 1,012 articles were excluded in this step. Finally, 190 articles fulfilled the criteria and were included in this review. These papers were published within 2000-2017. A total of 190 plants were included in this study. Based on literature search, 172 experimental studies and 71 clinical cases were included.Overall, results indicate that grape, soybean, green tea, garlic, olive, and pomegranate are the most effective plants against colon cancer. In these studies, fruits, seeds, leaves, and plant roots were used forin vitro and in vivo studies.


### 3.1.1.In Vitro Studies

Out of 172 studies, 75 were carried out on HT-29, 60 on HCT116, and 24 on Caco-2 cells (Table1). On HT-29 cells, both Allium sativum root extracts and Camellia sinensis leaf extracts induced cell apoptosis by two different mechanisms, respectively. In fact, the former showed inhibition of the PI3K/Akt pathway, upregulation of PTEN, and downregulation of Akt and p-Akt expression, while the latter was involved in attenuation of COX-2 expression and modulation of NFκB, AP-1, CREB, and/or NF-IL-6. Moreover, an antiproliferative activity has also been detected in Olea europaea fruit extracts, which increased caspase 3-like activity and were involved in the production of superoxide anions in mitochondria. An antiproliferative activity, by means of a blockage in the G2/M phase, has also been reported in Caco-2 cells by Vitis vinifera fruit extracts. Concerning HCT116 cells, several plants, such as American ginseng and Hibiscus cannabinus, induced cell cycle arrest in different checkpoints.Table 1
Cytotoxic effects of medicinal plants on colon cancer inin vitro models.
Scientific name
Parts used
Cell line
Conc.
Type of extract
Important compounds
Cellular effect
Mechanisms
References
Vitis vinifera
Fruit
HCT116
NM
Lyophilized
Hydroxycinnamic acids, proanthocyanidins, stilbenoids
Increase of dihydroceramides, sphingolipid mediators involved in cell cycle arrest, and reduction of the proliferation rate
(i) Increase of p53 and p21 cell cycle gate keepers(ii) Activation of the transcriptional factor Nrf2
[26, 27]
Fruit
Caco-2
365 mg/g
Methanolic
Catechin, epicatechin, quercetin, gallic acid
Antiproliferative activity and direct initiation of cell death
Blockage in the G2/M phase
[28, 29]
Seed
Caco-2
10–25μg/mL
Aqueous
Procyanidins
(i) Increased crypt depth(ii) Inhibited cell viability and decreased histological damage score
Reduced MPO (myeloperoxidase) activity
[29]
Skin
NM
7.5, 30, 60μg/mL
Methanolic
4′-Geranyloxyferulic acid
NM
NM
[30]
Seed
Colon cancer stem cells
6.25, 12.5, 25μg/mL
NM
(+)-catechin, (−)-epicatechin
NM
(i) Increment of p53, Bax/Bcl-2 ratio, and cleaved PARP(ii) Inhibition of Wnt/β-catenin signaling
[31]
Allium sativum
Root
HT-29
20, 50, 100 mg/mL
Ethanolic
NM
Induction of apoptosis and cell cycle arrest
(i) Inhibition of the PI3K̸Akt pathway(ii) Upregulation of PTEN and downregulation of Akt and p-Akt expression
[32]
Glycine max
Seed
Caco-2, SW620, HT-29
12.5μg/mL
Aqueous
Anthoxanthin
Cell death and significant reduction of cell density
Enhancement of Rab6 protein levels
[33]
Seed
HT-29
240, 600 ppm
Crude
Saponin
Suppression of PKC activation and increase of alkaline phosphatase activity
[33]
Seed
HT-29
NM
Crude
Saponin
NM
(i) Suppression of the degradation of IκBα in PMA-stimulated cells(ii) Downregulation of COX-2 and PKC expressions
[34]
Camellia sinensis
Leaf
HT-29
0, 10, 30, 50μM
Aqueous
Catechin, epigallocatechin gallate
1.9-fold increase in tumor cell apoptosis and a 3-fold increase in endothelial cell apoptosis
(i) Suppression of ERK-1 and ERK-2 activation(ii) Suppression of VEGF expression
[35]
Leaf
Caco-2, HT-29
300μM
Aqueous
Theaflavins (TF-2T, F-3, TF-1)
Human colon cancer cell apoptosis induction
Modulation of NFκB, AP-1, CREB, and/or IL-6
[36]
Leaf
HT-29
68-800.73 μg/mL
Hot water extract
Flavan-3-ol (catechin & tannin) & polyphenols (teadenol B)
Inhibition of proliferation of HT-29 cells
Increased expression levels of caspases 3/7, 8, and 9
[35]
Olea europaea
Fruit
HT-29
150, 55.5200 and 74 μmol/L
Methanolic and chloroform
Maslinic acid, oleanolic acid
Antiproliferative activity
(i) Increased caspase 3-like activity to 6-fold(ii) Production of superoxide anions in the mitochondria
[37]
Fruit, leaf
SW480 and HT-29
100–400 m/z
Methanolic & hexane
Oleic acid, linoleic acid, gamma-linolenic acid, lignans, flavonoids, secoiridoids
Reduced cell growth in both cell lines
(i) Limited G2M cell cycle(ii) Depressed cyclooxygenase-2 expression in HT-29 cells(iii) Suppression of β-catenin/TCF signaling in SW480 cells(iv) Promotion of the entry into subG1 phase
[38]
Fruit
Caco-2
50μM
Aqueous
Phenolic compounds, authentic hydroxyl tyrosol (HT)
Reduced proliferation of Caco-2 cells
Reduction of the methylation levels of CNR1 promoter
[39]
Fruit
HT115
25μg/mL
Hydroethanolic
Phenolic compounds (p-hydroxyphenyl ethanol, pinoresinol & dihydroxyphenyl ethanol)
NM
Inhibition by reduced expression of a range ofα5 & β1
[40]
Olive mill wastewater
HT-29, HCT116, CT26
NM
Methanolic
Hydroxytyrosol
(i) Inhibited proliferation(ii) Inhibited migration and invasion
(i) Reduced sprout formation(ii) Inhibited VEGF and IL-8 levels
[41]
Fruit
Caco-2
0-2,000μg/mL
Ethanolic
Tyrosol, hydroxytyrosol, oleuropein, rutin, quercetin and glucoside forms of luteolin and apigenin
NM
(i) Induction of the cell cycle arrest in S-phase
[42]
Punica granatum
Juice
HT-29
50 mg/L
Aqueous
Ellagitannins, punicalagin
Inhibition of cancer cell proliferation
(i) Suppressed TNFR-induced COX-2 protein expression(ii) Reduced phosphorylation of the p65 subunit and binding to the NFκB response element
[43]
Seed
LS174
63.2μg/mL
Supercritical fluid
Punicic acid,γ-tocopherol, α-tocopherol
Cytotoxic activity
(i) Slightly decreased development of tubules from elongated cell bodies(ii) Reduction of the number of cell connections
[44]
Glycyrrhiza glabra
Root
HT-29
12.2 and 31μg/mL
Ethanolic
Licochalcone
NM
Increase of the protein levels of proapoptotic Bax
[37]
Opuntia ficus-indica
Fruit
Caco-2
115μM
Aqueous
Betalain pigment indicaxanthin
Apoptosis of proliferating cells
(i) Demethylation of the tumor suppressor p16INK4a gene promoter(ii) Reactivation of the silenced mRNA expression and accumulation of p16INK4a
[38]
Fruit
HT-29 & Caco-2 & NIH 3 T3 (as control)
Against HT-29 (4.9μg/mL) against Caco-2 (8.2 μg/mL)
Alkaline hydrolysis with NaOH
Isorhamnetin glycosides (IG5 and IG6)-phenol
Cell death through apoptosis and necrosis
Increased activity of caspase 3/7
[45]
Piper betle
Leaf
HT-29 and HCT116
200.0μg/mL
Aqueous
Hydroxychavicol
Antioxidant capacity and induction of a greater apoptotic effect
(i) Scavenging activity(ii) Formation of electrophilic metabolites
[41, 46]
Fragaria×ananassa
Fruit
HT-29
0.025, 0.05, 0.25, 0.5%
Ethanolic
Ascorbate, ellagic acid
Decreased proliferation of HT-29 cells
Increase in the levels of 8OHA and decrease in the levels of 8OHG
[40]
Sasa quelpaertensis
Leaf
HT-29 HCT116
0, 100, 200, 300 mg/L
Ethanolic
p-Coumaric acid, tricin
Inhibited colony formation
Nonadherent sphere formation suppressed CD133+ & CD44+ population
[41]
Salvia chinensis
Stem
HCT116, COLO 205
10, 20, 40,60, 80, 100 mg/L
Polyphenolic
Terpenoids, phenolic acid, flavonoids, dibenzylcyclooctadiene
Apoptosis & loss of mitochondrial membrane
Induced G0/G1 cell cycle
[42]
Rubus idaeus L.
Fruit
HT-29, HT-115, Caco-2
3.125, 6.25, 12.5, 25, 50 mg/L
Acetate
Polyphenol, anthocyanin, ellagitannin
NM
Decreased population of cells in G1 phase
[47]
LoVo
50μL
Aqueous
NM
Inhibited proliferation of LoVo
Suppression of the NFκB pathway
[48]
Curcuma longa
Root
HT-29, HCT15, DLD1, HCT116
(i) Short-term assay: four 10-fold dilutions (100 to 0.1 mg/L)(ii) Long-term assay: 5, 10, 20 mg/L
Ethanolic
Curcumin (diferuloylmethane)
Inhibited formation of HCT116 spheroids
NM
[49]
Eleutherococcus senticosus
Root
HCT116
12.5, 25, 50, 100
Methanolic
Eleutherosides, triterpenoid saponins, glycans
NM
Activation of natural killer cells and thus enhancement of immune function
[50]
Tabernaemontana divaricata L.
Leaf
HT-29, HCT15
10, 30, 100 mg/L
Ethyl acetate, chloroform, methanolic
Alkaloids
NM
Inhibited the unwinding of supercoiled DNA
[45]
Millingtonia hortensis
Root, flower, leaf
RKO
50, 100, 200, 400, 800 mg/L
Aqueous
Phenylethanoid glycoside, squalene, salidroside, 2-phenyl rutinoside
Apoptosis induction
(i) Increase of fragmented DNA(ii) Decrease of the expression of antiapoptotic proteins, Bcl-xL and p-BAD
[46]
Powder
RKO
200, 400, 800μg/mL
Aqueous
Water soluble compounds
Antiproliferative effect
NM
[51]
Thai purple rice
Seed
Caco-2, Cat. No. HTB-37
16.11μg/mL
Methanol acidified
Cyanidin-3-glucoside and peonidin-3-glucoside, anthocyanins, phenolic compounds
(i) Antioxidation of anthocyanins and phenols(ii) Antiproliferation of colon cancer cells
NM
[52]
Annona muricata
Leaf
HCT116, HT-29
11.43±1.87μg/ml and 8.98±1.24μg/ml
Ethanolic
Alkaloids, acetogenins, essential oils
Block of the migration and invasion of HT-29 and HCT116 cells
(i) Cell cycle arrest at G1 phase(ii) Disruption of MMP, cytochrome c leakage and activation
[53]
NM
HT-29, HCT116
<4, <20μg/mL
EtOAc
Annopentocin A, annopentocin B, annopentocin C, cis- and trans-annomuricin-D-ones, annomuricin E
NM
Suppression of ATP production and NADH oxidase in cancer cells
[54]
Pistacia lentiscus L. var. chia
Leaf
HCT116
NM
Ethanolic
Resin, known as Chios mastic gum (CMG)
Causes several morphological changes typical of apoptosis in cell organelles
(i) Induction of cell cycle arrest at G1 phase(ii) Activation of pro-caspases 8, 9, 3
[55]
Resin
HCT116
100μg/mL
Hexane
Caryophyllene
Induction of the anoikis form of apoptosis in human colon cancer HCT116 cells
(i) Induction of G1 phase arrest(ii) Loss of adhesion to the substrate
[56]
American ginseng (Panax quinquefolius)
Biological constituents
HCT116
0-2.0 mg/mL
Aqueous
Ginseng (GE) or its ginsenoside (GF) and polysaccharide (PS)
Proliferation was inhibited by GE, GF, and PS in wild-type and p21 cells
(i) Cells arrest in G0/G1 phase and increment of p53 and p21 proteins(ii) Increment of Bax and caspase 3 proteins expression
[57]
Purple-fleshed potatoes
Fruit
Colon cancer stem cells
5.0μg/mL
Ethanol, methanol, ethyl acetate
Anthocyanin,β-catenin, cytochrome c
Critical regulator of CSC proliferation and its downstream proteins (c-Myc and cyclin D1) and elevated Bax and cytochrome c
(i) Cytochrome c levels were elevated regardless of p53 status(ii) Mitochondria-mediated apoptotic pathway(iii) Suppressed levels of cytoplasmic and nuclear β-catenin
[58]
Phaseolus vulgaris
Leaf
HT-29
NM
Ethanolic
Polysaccharides, oligosaccharides
Changes in genes involved or linked to cell cycle arrest
(i) Inactivation of the retinoblastoma phosphoprotein(ii) Induction of G1 arrest(iii) Suppression of NF-jB1(iv) Increase in EGR1 expression
[59]
Opuntia spp.
Fruit
HT-29
5.8±1.0, 7.5±2.0, 12±1% (V/V)
Hydroalcoholic
Betacyanins, flavonoids (isorhamnetin derivatives) and phenolic acids (ferulic acid)
NM
Induced cell cycle arrest at different checkpoints—G1, G2/M, and S
[60]
Suillus luteus
NM
HCT15
400μg/mL
Methanolic
Protocatechuic acid, cinnamic acid,α-tocopherol, β-tocopherol, mannitol, trehalose, polyunsaturated fatty acids, monounsaturated fatty acids, saturated fatty acids
(i) Increase in the cellular levels of p-H2A.X, which is suggestive of DNA damage
(i) Inhibition of cell proliferation in G1 phase(ii) Increase in the cellular levels of p-H2A.X
[61]
Poncirus trifoliata
Leaf
HT-29
0.63μM
Aqueous (in acetone)
β-Sitosterol, 2-hydroxy-1,2,3-propanetricarboxylic acid 2-methyl ester
Arrest of cell growth was observed withβ-sitosterol
NM
[62]
Rosmarinus officinalis L.
Leaf
SW 620, DLD-1
0-120μg/mL
Methanolic
Polyphenols
Antiproliferative effect of 5-FU
Downregulation of TYMS and TK1, enzymes related to 5-FU resistance
[63]
Leaf
HT-29
SC-RE 30μg/mL and CA 12.5 μg/mL
Ethanolic
Polyphenols (carnosic acid (CA) and carnosol)
(i) Upregulation of VLDLR gene as the principal contributor to the observed cholesterol accumulation in SC-RE-treated cells(ii) Downregulation of several genes involved in G1-S
Activation of Nrf2 transcription factor and common regulators, such as XBP1 (Xbp1) gene related to the unfolded protein response (UPR)
[64]
NM
HT-29
10, 20, 30, 40, 50, 60, 70μg/mL
NM
Carnosic acid, carnosol, rosmarinic acid, rosmanol
NM
NM
[65]
Leaf
HGUE-C-1, HT-29, and SW480
20–40 mg/mL
CO2-supercritical fluid extract
Carnosic acid, carnosol, and betulinic acid
NM
(i) Prooxidative capability by increasing the intracellular generation of ROS(ii) Activation of Nrf2
[66]
Glehnia littoralis
Leaf
HT-29
50 mg/mL
Methanolic
Bergapten, isoimpinellin, xanthotoxin, imperatorin, panaxydiol, falcarindiol, falcarinol
Induced apoptosis by the decreased expression of the antiapoptotic Bcl-2 mRNA
(i) Reduced expression of Bcl-2(ii) Reduced expression levels of iNOS and COX-2
[67]
Verbena officinalis
Leaf
HCT116
20 mg/mL
Aqueous
Phenylethanoid glycosides, diacetyl-O-isoverbascoside, diacetyl-O-betonyoside A, and diacetyl-O-betonyoside A
(i) Substantial tumor cell growth inhibitory activity(ii) Time-dependent cytotoxicity against both cell lines
(i) Increased lipophilicity of molecules seemed to be responsible for enhanced cytotoxicity(ii) Antiproliferative activity is determined by the number of acetyl groups and also by their position in the aliphatic rings
[68]
Mentha spicata
Leaf
RCM-1
12.5μg/mL
N-Hexane
Acetic acid 3-methylthio propyl ester (AMTP), methyl thio propionic acid ethyl ester (MTPE)
Exhibited antimutagenic activity
Auraptene (7-geranyloxycoumarin) having a monoterpene moiety andβ-cryptoxanthin (one of the tetraterpenes) increased antibody production
[69]
Euphoria longana Lam.
Seed
SW 480
0–100μg/mL
Ethanolic
Corilagin, gallic acid, ellagic acid
(i) Antiangiogenetic properties(ii) All fractions showed the anti-VEGF secretion activity
Release and expression of VEGF indicated that all fractions showed the anti-VEGF secretion activity
[70]
Sutherlandia frutescens
Flower
Caco-2
1/50 dilution of the ethanolic extract
Ethanolic
Amino acids, including L-arginine and L-canavanine, pinitol, flavonoids, and triterpenoid saponins as well as hexadecanoic acid andγ-sitosterol
Disruption of the key molecules in the PI3K pathway thereby inducing apoptosis
Decrease in cell viability and increment in pyknosis as well as loss in cellular membrane integrity
[71]
Melissa officinalis
Leaf
HT-29, T84
346, 120μg/mL
Ethanolic
Phenolic acids (rosmarinic acid, coumaric acid, caffeic acid, protocatechuic acid, ferulic acid, chlorogenic acid), flavonoids, sesquiterpenes, monoterpenes, triterpenes
(i) Inhibited proliferation of colon carcinoma cells(ii) Induced apoptosis through formation of ROS
(i) G2/M cell cycle arrest(ii) Cleavage of caspases 3 and 7(iii) Induced phosphatidylserine externalization in colon carcinoma cells(iv) Induced formation of ROS in colon carcinoma cells
[72]
Sargassum cristaefolium
Leaf
HT-29
500 mg/mL
Ethanolic
Fucoidans
(i) Reduction of free radicals(ii) DPPH radical scavenging
Accumulation of cells in G0/G1 phase
[73]
Hedyotis diffusa
NM
HT-29
400 mg/mL
Ethanolic and then DMSO
Octadecyl (E)-p-coumarate, P-E-methoxy-cinnamic acid, ferulic acid, scopoletin, succinic acid, aurantiamide acetate, rubiadin
Suppress tumor cell growth and induce the apoptosis of human CRC cells
(i) Block G1/S progression(ii) Induce the activation of caspases 9 and 3(iii) Inhibit IL-6-mediated STAT3 activation(iv) Downregulate the mRNA and protein expression levels of cyclin D1, CDK4, Bcl-1, and Bax
[74]
Zingiber officinale Roscoe
Peel
LoVo
100 mg/mL
Ethanolic
Linoleic acid methyl ester,α-zingiberene, and zingiberone
Interesting antiproliferative activity against colorectal carcinoma
NM
[75]
Scutellaria barbata
Leaf
LoVo
413.3 mg/L
Methanolic
Scutellarein, scutellarin, carthamidin, isocarthamidin, wogonin
Induce cell death in the human colon cancer cell line
Increase in the sub-G1 phase and inhibition of cell growth
[76]
Pistacia atlantica, Pistacia lentiscus
Resin
HCT116
100μg/mL
Hexane extract
Caryophyllene
Induce the anoikis form of apoptosis in human colon cancer HCT116 cells
(i) Induce G1 phase arrest(ii) Loss of adhesion to the substrate
[56]
Citrus reticulata
Peel
SNU-C4
100μg/mL
Methanolic
Limonene, geranial, neral, geranyl acetate, geraniol,β-caryophyllene, nerol, neryl acetate
Induce the apoptosis on SNU-C4, human colon cancer cells
Expression of proapoptotic gene, Bax, and major apoptotic gene, caspase 3
[77]
Echinacea pallida, Echinacea angustifolia, Echinacea purpurea
Root
COLO320
150 mg/mL
Hexanic
Caffeic acid derivatives, alkylamides, polyacetylenes, polysaccharides
Induce apoptosis and promote nuclear DNA fragmentation
(i) Induce apoptosis by increasing caspase 3/7 activity(ii) Promote nuclear DNA fragmentation
[78]
Nasturtium officinale
Leaf
HT-29
50μL/mL
Methanolic
Phenethyl isothiocyanate, 7-methylsulfinylheptyl, 8-methylsulfinyl
(i) Inhibition of initiation, proliferation, and metastasis
(i) Inhibited DNA damage(ii) Accumulation of cells in S phase of the cell cycle
[79]
Polysiphonia
NM
SW480, HCT15, HCT116, DLD-1
20 and 40μg/mL
Methanolic
2,5-Dibromo-3,4-dihydroxybenzyl n-propyl ether
Potentially could be used as a chemopreventive agent against colon cancer
(i) Inhibited Wnt/β-catenin pathway(ii) Repressed CRT in colon cancer cells(iii) Downregulated cyclin D1(iv) Activated the NFκB pathway
[80]
Aristolochia debilis Sieb. et Zucc.
Stem
HT-29
200μg/mL
Methanolic
Aristolochic acid, nitrophenanthrene carboxylic acids
Inhibition of proliferation and induction of apoptosis in HT-29 cells
(i) Induction of sub-G1 cell cycle(ii) Generation of ROS and decrease of the MMP(iii) Bax overexpression and increase of Bax/Bcl-2 ratio
[81]
Myrtaceae
Leaf
HCT116
100μg/mL (in vitro), 200 and 100 μg/disc (in vivo)
Methanolic
Phenols, flavonoid, betulinic acid
Strong inhibition of microvessel outgrowth
(i) Inhibition of tube formation on Matrigel matrix(ii) Inhibition of HUVECS migration (in vitro)(iii) Decreased nutrient and oxygen supply
[82]
Spica prunellae
Leaf
HT-29
200 mg/mL (in vitro), 600 mg/mL (in vivo)
Ethanolic
Rosmarinic acid
Inhibits CRC cell growth
(i) Suppresses STAT3 phosphorylation(ii) Regulates the expression of Bcl-2, Bax, cyclin D1, CDK4, VEGF-A, and VEGFR-2
[83]
Phytolacca americana
Root
HCT116
3200μg/mL
Ethanolic
Jaligonic acids, kaempferol, quercetin, quercetin 3-glucoside, isoquercitrin, ferulic acid
Control of growth and spread of cancer cells
Reduction in the expressions of MYC, PLAU, and TEK
[84]
Morus alba
Leaf
HCT15
13.8μg/mL
Methanolic
Epicatechin, myricetin, quercetin hydrate, luteolin, kaempferol, ascorbic acid, gallic acid, pelargonidine, p-coumaric acid
Cytotoxic effect on human colon cancer cells (HCT15)
(i) Apoptosis induction also involved in the downregulation of iNOS(ii) Fragmentation of DNA(iii) Upregulation of caspase 3 activity
[85]
Rhodiola imbricata
Leaf
HT-29
200μg/mL
Acetone and methanolic
Phenols, tannins, and flavonoids
(i) Antioxidant activity(ii) Inhibited proliferation of HT-29 cells
(i) Scavenge free radicals(ii) DPPH radical scavenging activity(iii) Increased metal chelating activity
[86]
Asiasarum heterotropoides F.
DriedA. radix
HCT116
20 mg/mL
Ethanolic
Asarinin and xanthoxylol
Inhibition of the growth of HCT116 cells
(i) Caspase-dependent apoptosis(ii) Regulation of p53 expression at transcription level
[87]
Podocarpus elatus
Fruit
HT-29
500 mg/mL
Methanolic
Phenolic and anthocyanin
Reduction of proliferation of colon cancer cells
(i) Cell cycle delay in S phase(ii) 93% downregulation of telomerase activity and decrease in telomere length(iii) Induced morphological alterations to HT-29 cells
[88]
Echinacea purpurea
Flower
Caco-2, HCT116
0–2,000 mg/mL
Hydroethanolic
Cichoric acid
(i) Inhibition of proliferation(ii) Decreased telomerase activity in HCT116 cells
(i) Decreased telomerase activity(ii) Activation of caspase 9(iii) Cleavage of PARP(iv) Downregulation of β-catenin
[89]
Root
COLO320
150 mg/mL
Hexanic
Caffeic acid derivatives, alkylamides, polyacetylenes, polysaccharides
Induce apoptosis by increasing significantly caspase 3/7 activity and promote nuclear DNA fragmentation
(i) Increase significantly caspase 3/7 activity(ii) Promote nuclear DNA fragmentation
[78]
Hop (Humulus lupulus L.), Franseria artemisioides
Leaf
NM
100 mg/kg b.w./day
Aqueous
Coumarin, lignans, quinones
30% reduction of tumor-induced neovascularization
NM
[90]
NM
Caco-2
NM
Ethanolic
Phenolic compounds, flavonoid, diterpenes
Digestive, gastroprotective, antiseptic, anti-inflammatory, and antiproliferative activity
NM
[91]
Fruit
NL-17
0, 50, 100, 150μg/mL
Methanolic
α-Mangostin (xanthone)
NM
(i) Induction of caspase 3 and caspase 9 activation(ii) Induced cell cycle arrest at G1/G0 phase
[92]
Stem, bark
HT-29
50μg/mL
Chloroform-soluble
β-Mangostin, garcinone D, cratoxyxanthone
Cytotoxic activity against HT-29 human colon cancer
Inhibition of p50 and p65 activation
[93]
Annona squamosa Linn
Leaf
HCT116
8.98μg/mL
Crude, Aq ethyl acetate
Acetogenins (annoreticuin & isoannoreticuin) and alkaloids dopamine, salsolinol, and coclaurine
Inhibition of growth and proliferation of tumor cells
(i) Reactive oxygen species (ROS) formation, lactate dehydrogenase (LDH) release(ii) Activation of caspases 3/7, 8, and 9
[94]
Derris scandens
Stem
HT-29
5-15μg/mL
Ethanolic
Benzyls and isoflavones (genistein, coumarins, scandinone)
Apoptosis and mitotic catastrophe of human colon cancer HT-29 cells
(i) Inhibition ofα-glucosidase activity(ii) Scavenge free radicals
[95]
Eupatorium cannabinum
Aerial parts
HT-29
25μg/mL
Ethanolic
Pyrrolizidine alkaloids (senecionine, senkirkine, monocrotaline, echimidine)
Induced alteration of colony morphology
(i) Upregulation of p21 and downregulation of NCL, FOS, and AURKA(ii) Mitotic disruption and nonapoptotic cell death via upregulation of Bcl-xL, limited TUNEL labeling, and nuclear size increase
[96]
Sorghum bicolor
The dermal layer of stalk
HCT116 & colon cancer stem cells
>16 and 103μg/mL
Phenolic-rich ethanolic, acetone
Apigeninidin & luteolinidin
Antiproliferative
Target p53-dependent and p53-independent pathways
[97]
Dermal and seed head
CCSC
NM
Methanolic
Apigeninidin, luteolinidin, malvidin 3-O-glucoside, apigenin, luteolin, naringenin, naringenin 7-O-glucoside, eriodictyol 5-glucoside, taxifolin, catechins
NM
(i) Elevation of caspase 3/7 activity(ii) Decrease in β-catenin, cyclin D1, c-Myc, and survivin protein levels(iii) Suppression of Wnt/β-catenin signaling in a p53-dependent (dermal layer) and partial p53-dependent (seed head) manner
[98]
Hibiscus cannabinus
Seed
HCT116
KSE (15.625μg/mL to 1,000 μg/mL)
Ethanolic
Gallic acid, p-hydroxybenzoic acid, caffeic acid, vanillic acid, syringic acid, and p-coumaric and ferulic acids
Cytotoxic activity against human colon cancer HCT116 cells
Apoptosis via blockade of mid G1-late G1-S transition thereby causing G1 phase cell cycle arrest
[99]
Salix aegyptiaca L.
Bark
HCT116 & HT-29
300μg/mL
Ethanolic
Catechin, salicin, catechol and smaller amounts of gallic acid, epigallocatechin gallate (EGCG), quercetin, coumaric acid, rutin, syringic acid, and vanillin
Anticarcinogenic effects in colon cancer cells
Apoptosis via inhibition of phosphatidylinositol 3-kinase/protein kinase B and mitogen-activated protein kinase signaling pathways
[100]
Rubus coreanum
Fruit
HT-29
400μg/mL
Aqueous
Polyphenols, gallic acid, sanguine
Induction of apoptosis
(i) Induced activity of caspases 3, 7, and 9(ii) Cleavage of poly(adenosine diphosphate-ribose) polymerase
[101]
Codonopsis lanceolata
Root
HT-29
200μg/mL
N-Butanol fraction
Tannins, saponins, polyphenolics, alkaloids
Apoptosis in human colon tumor HT-29 cells
(i) Induced G0/G1 arrest(ii) Enhancement of expression of caspase 3 and p53 and of the Bax/Bcl-2 ratio
[102]
Gleditsia sinensis
Thorn
HCT116
800μg/mL
Aqueous
Flavonoid, lupine acid, ellagic acid glycosides
(i) Increase in p53 levels(ii) Downregulation of the checkpoint proteins, cyclin B1, Cdc2, and Cdc25c
Inhibition of proliferation of colon cancer cells
[90]
Thorn
HCT116
600μg/mL
Ethanolic
NM
Inhibitory effect on proliferation of human colon cancer HCT116 cells
(i) Caused cell cycle arrest at G2/M phase together with a decrease of cyclin B1 and Cdc2(ii) Progression from G2/M phase
[91]
Ligustrum lucidum
Fruit
DLD-1
50μg/mL
Aqueous
Oleanolic acid, ursolic acid
Inhibited proliferation
(i) Reduction of Tbx3 rescued the dysregulated P14ARF-P53 signaling
[94]
Zingiber officinale
Rhizome
HCT116
5μM
Ethanolic
6-Paradol, 6- and 10-dehydrogingerdione, 6- and 10-gingerdione, 4-, 6-, 8-, and 10-gingerdiol, 6-methylgingerdiol, zingerone, 6-hydroxyshogaol, 6-, 8-, 10-dehydroshogaol, diarylheptanoids
Inhibitory effects on the proliferation of human colon cancer cells
(i) Arrest at G0/G1 phase(ii) Reduced DNA synthesis
[103]
Grifola frondosa
Fruit
HT-29
10 ng/mL
Aqueous
Phenolic compounds (pyrogallol, caffeic acid, myricetin, protocatechuic acid)
Inhibition of TNBS-induced rat colitis
Induced cell cycle progression in G0/G1 phase
[104]
Cucumaria frondosa
The enzymatically hydrolyzed epithelium of the edible
HCT116
<150μg/mL
Hydroalcoholic
Monosulphated triterpenoid glycoside frondoside A, the disulphated glycoside frondoside B, the trisulphated glycoside frondoside C
Inhibition of human colon cancer cell growth
(i) Inhibition at S and G2-M phases with a decrease in Cdc25c and increase in p21WAF1/CIP(ii) Apoptosis associated with H2AX phosphorylation and caspase 2
[105]
Rolandra fruticosa
Leaf & twigs
HT-29
10 and 5 mg/kg/day
Methanolic
Sesquiterpene lactone (13-acetoxyrolandrolide)
Antiproliferative effect against human colon cancer cells
Inhibition of the NFκB pathway, NFκB subunit p65 (RelA), upstream mediators IKKβ and oncogenic K-ras
[106]
Cydonia oblonga Miller
Leaf & Fruit
Caco-2
250–500μg/mL
Methanolic
Phenolic compound (flavonol and flavone heterosides, 5-O-caffeoylquinic acid)
Antiproliferative effect against human kidney and colon cancer cells
(i) Suppression of factor activation, nuclear factor-kB (NFκB) activation, protein-1 (AP-1) transcription factor, mitogen protein kinases (MAPKs), protein kinases (PKs), namely, PKC, growth-factor receptor- (GFR-) mediated pathways and angiogenesis(ii) Cell cycle arrest and induction of apoptosis, antioxidant, and anti-inflammatory effects
[107]
Morchella esculenta
Fruits
HT-29
820 mg/mL
Methylene chloride
Steroids (mainly ergosterol derivatives) & polysaccharides & galactomannan
Antioxidant activity in HT-29 colon cancer cells
Inhibition of NF-B activation in the NF-B assay
[108]
Sedum kamtschaticum
Aerial part
HT-29
0–0.5 mg/mL
Methanolic
Buddlejasaponin IV
Induced apoptosis in HT-29 human colon cancer cells
Induction of apoptosis via mitochondrial pathway by downregulation of Bcl-2 protein levels, caspase 3 activation, and subsequent PARP cleavage
[109]
Ginseng and Glycyrrhiza glabra
Leaf
HT-29
500μL
Aqueous
Uracil, adenine, adenosine, Li-glycyrrhetinic acid, quiritin
NM
Antiproliferative effect determination of the protein levels of p21, cyclin D1, PCNA, and cdk-2, which are the key regulators for cell cycle progression
[110]
Orostachys japonicus
Leaf & stem
HT-29
2 mg/mL
Aqueous
Flavonoids, triterpenoids, 4-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, polysaccharide
Antiproliferation in HT-29 colon cancer cells
Inhibited proliferation at G2 point of the cell cycle and apoptosis via tumor suppressor protein p53
[111]
Ginkgo biloba
Fruit & leaf
HT-29
20–320mg/L
Aqueous
Terpene lactones and flavonoid glycosides
(i) Inhibited progression of human colon cancer cells(ii) Induced HT-29 cell apoptosis
Increase in caspase 3 activities and elevation in p53 MRN reduction in Bcl-2 mRNA
[112]
Oryza sativa
Seed
HT-29, SW 480, HCEC
100μg/mL
Ethyl acetate
Phenolic compound (tricin, ferulic acid, caffeic acid, and methoxycinnamic acid)
Inhibition of the human colon cancer cell growth
(i) Induced apoptosis by enhanced activation of caspases 8 and 3(ii) Decrease of the number of viable SW480 and HCEC cells(iii) Reduced colony-forming ability of these cells
[113]
Cnidium officinale Makino
Root
HT-29
305.024/mL
Ethanolic
Osthole, auraptenol, imperatorin
Inhibited proliferation of human colon cancer cells (HT-29)
Inhibition of the cellular proliferation via G0/G1 phase arrest of the cell cycle and induced apoptosis
[114]
Cnidium officinale Makino
Root
HT-29
0.1-5 mg/mL
Aqueous
N-(3-(Aminomethyl)benzyl)acetamidine
Inhibited the invasiveness of cytokine-treated HT-29 cells through the Matrigel-coated membrane in a concentration-dependent manner
(i) Reduction of HT-29 cell invasion through the Matrigel(ii) Inhibited cytokine-mediated NO production, iNOS expression, and invasiveness of HT-29 cells(iii) Inhibited MMP-2 activity
[115]
Long pepper (PLX)
Fruit
HT-29 and HCT116
0.10 mg/mL
Ethanolic
Piperidine alkaloids, piperamides, piperlongumine
(i) Induction of apoptosis, following DNA fragmentation in HT-29 colon cancer cells in a time-dependent manner(ii) Induced caspase-independent apoptosis
Induced whole cell ROS production
[116]
Achyranthes aspera
Root
COLO 205
50-100 and 150-200μg/mL
Ethanolic (EAA) and aqueous (AAA) root extractsAqueous
Phenolic compounds
(i) Enhanced growth inhibitory effects of AAA towards COLO 205 cells in contrast to EAA(ii) Stimulatory role of AAA in the activation of cell cycle inhibitors
(i) Triggered mitochondrial apoptosis pathway and S phase cell cycle arrest(ii) Increased levels of caspase 9, caspase 3, and caspase 3/7 activity
[117]
Thymus vulgaris
Leaf
HCT116
0.2, 0.4, 0.6, 0.8 mg/mL
Carvacrol and thymol
Inhibited proliferation, adhesion, migration, and invasion of cancer cells
[118]
Dictyopteris undulata
NM
SW480
40μg/mL
Ethanolic
Cyclozonarone benzoquinone
NM
Induced apoptosis by reducing Bcl-2 levels, upregulating Bax, and disrupting the mitochondrial membrane potential, leading to the activation of caspases 3 and 9
[119]
Dendrobium microspermae
NM
HCT116
0.25, 0.5, 1.0 mg/mL
Methanolic
NM
NM
Upregulation of Bax and caspases 9 and 3 and downregulation of Bcl-2 expression of genes
[120]
Cannabis sativa
Dry flower & leaf
DLD-1 and HCT116
0.3–5μM
Methanolic
Cannabidiol, phytocannabinoids
Reduced cell proliferation in a CB1-sensitive
(i) Reduced AOM-induced preneoplastic lesions and polyps(ii) Inhibited colorectal cancer cell proliferation via CB1 and CB2 receptor activation
[121]
Phoenix dactylifera L.
Fruit
Caco-2
0.2 mg/mL
Aqueous
Phenolic acids (gallic, protocatechuic, hydroxybenzoic, vanillic, isovanillic, syringic, caffeic, ferulic, sinapic, p-coumaric, isoferulic), flavonoid glycosides (quercetin, luteolin, apigenin, and kaempferol), and anthocyanidins
Increasing beneficial bacterial growth and inhibition of proliferation of colon cancer cells
NM
[122]
Melia toosendan
Fruit
SW480, CT26
0, 10, 20, 30, 40, 50μg/mL
Ethanolic
Triterpenoids, flavonoids, polysaccharide, limonoids
NM
(i) Inhibited cell proliferation of SW480 and CT26 by promoting apoptosis as indicated by nuclear chromatin condensation and DNA fragmentation(ii) Induced caspase 9 activity which further activated caspase 3 and poly(ADP-ribose) polymerase cleavage, leading the tumor cells to apoptosis
[123]
Crocus sativus L.
Flower
HCT116
0.25, 0.5, 1, 2, 4μg/mL
Ethanolic
Carotenoid, pigment, crocin, crocetin
Induced DNA damage and apoptosis
(i) Induction of a p53 pattern-dependent caspase 3 activation with a full G2/M stop(ii) Induced remarkable delay in S/G2 phase transit with entry into mitosis
[124]
Tepals and leaf
Caco-2
0.42 mg/mL
NM
Polyphenols, glycosides of kaempferol, luteolin, and quercetin
Proliferation of Caco-2 cells was greatly inhibited
NM
[125]
Luffa echinata
Fruit
HT-29
50, 100, and 200μg/mL
Methanolic
Amariin, echinatin, saponins, hentriacontane, gypsogenin, cucurbitacin B, datiscacin, 2-O-β-D-glucopyranosyl cucurbitacin B, and 2-O-β-D-glucopyranosyl cucurbitacin S
Increase in the population of apoptotic cells
(i) Inhibited the cellular proliferation of HT-29 cells via G2/M phase arrest of the cell cycle(ii) Induced apoptotic cell death via ROS generation(iii) Accumulation of caspase 3 transcripts of HT-29 cells
[126]
Vitis aestivalis hybrid
Fruits (wine)
CCD-18Co
25, 50, 100μg/mL
NM
Polyphenolics
NM
(i) Decreased mRNA expression of lipopolysaccharide- (LPS-) induced inflammatory mediators NFκB, ICAM-1, VCAM-1, and PECAM-1(ii) Enhanced expression of miR-126(iii) Decreased gene expression and reduced activation of the NFκB transcription factor, NFκB-dependent(iv) Decrease in ROS 113MAH
[127]
Xylopia aethiopica
Dried fruit
HCT116
0, 5, 10, 15, 20, 25, 30μg/mL
Ethanolic
Ent-15-oxokaur-16-en-19-oic acid (EOKA)
NM
(i) Induced DNA damage, cell cycle arrest in G1 phase, and apoptotic cell death
[128]
Sorghum
Grain
ER-β; nonmalignant young adult mouse colonocytes
1, 5, 10, 100μg/mL
Aqueous
Flavones (luteolin and apigenin), 3-deoxyanthocyanins naringenin (eriodictyol and naringenin)
Reduced cell growth via apoptosis
Increased caspase 3 activity
[129]
NM
HT-29, HCT116
0.9-2.0 mg/mL
Hydroethanolic
Procyanidin B1, delphinidin-3-O-glucoside, tannin, cyanidin-3-O glucoside
(i) Significantly arrested HT-29 cells in G1(ii) Highest growth inhibition(iii) Increased percentage of apoptotic cells
(i) Downregulation of apoptotic proteins, such as cIAP-2, livin, survivin, and XIAP, was seen in HCT116 cells(ii) Inhibition of tyrosine kinase
[130]
Panax notoginseng (Burk.) F.H. Chen
Root
LoVo and Caco-2
0, 100, 250, and 500μg/mL
Alcoholic
Saponin, ginsenoside
NM
Delay in progression of the G0/G1, S, or G2/M cell cycle phases
[131]
Brassica oleracea L. var. italica
Broccoli florets
HCT116
0, 1, 2.5, 5, 10μg/mL
Ethanolic
Glucoiberin, 3 hydroxy,4(α-L-rhamnopyranosyloxy), benzyl glucosinolate 4-vinyl-3-pyrazolidinone 4-(methyl sulphinyl), butyl thiourea, β-thioglucoside N-hydroxysulphates
NM
NM
[132]
Cistanche deserticola
Dried stem
SW480
In vivo: 0.4 g/kg/dayIn vitro: 100 μg/mL
Aqueous
Polysaccharides, phenylethanoid glycosides
(i) Decreased number of mucosal hyperplasia and intestinal helicobacter infection(ii) Increased number of splenic macrophage, NK cells, and splenic macrophages
Decreased frequency of hyperplasia andHelicobacter hepaticus infection of the intestine
[133]
Chaenomeles japonica
Fruit
Caco-2 and HT-29
10, 25, 50, 75, 100, 125, 150μM CE
NM
Procyanidins
NM
NM
[134]
Prunus mume
Fruit
SW480, COLO, and WiDr
150, 300, and 600μg/mL
Hydrophobic
Triterpenoid saponins
NM
(i) Inhibited growth and lysed SW480, COLO, and WiDr(ii) Induction of massive cytoplasmic vacuoles
[135]
Solanum lyratum
NM
COLO 205
50, 100, 200, 300, 400μg/mL
EtOH
β-Lycotetraosyl
Induced S phase arrest and apoptosis
(i) Induced DNA fragments(ii) Increased the levels of p27, p53, cyclin B1, active-caspase 3, and Bax(iii) Decreased the levels of Cdk1, pro-caspase 9, Bcl-2 and NF-ÎB, p65, and p50
[136]
Onopordum cynarocephalum
Aerial parts
HCT116, HT-29
0, 0.04, 0.12, 0.2, 0.4, 1.2 mg/mL0, 0.2, 0.4, 1.2, 2.0, 3.0 mg/mL
Aqueous
Flavonoids, lignans, and sesquiterpene lactones
NM
(i) Increase in the expression of proapoptotic proteins such as p53, p21, and Bax(ii) Inhibition of the antiapoptotic protein Bcl-2(iii) Decrease in cyclin D1 protein
[137]
Eleutherine palmifolia
Bulbs
SW480
2.5, 5, 10μg/mL
MeOH
Eleutherin, isoeleutherin
NM
(i) Inhibited the transcription of TCF/β-catenin(ii) Decrease in the level of nuclear β-catenin protein
[138]
Asparagus officinalis
Spears
HCT116
76μg/mL
Acetone
Steroidal saponins (HTSA-1, HTSAP-2, HTSAP-12, HTSAP-6, HTSAP-8)
NM
(i) Inhibition of Akt, p70S6K, and ERK phosphorylation(ii) Induction of caspase 3 activity, PARP-1 cleavage, DNA fragmentation, G0/G1 cell cycle arrest by reducing the expression of cyclins D, A, and E
[139]
Phyllanthus emblica L.
Seed, pulp
HCCSCs, HCT116
200μg/mL
Methanolic
Trigonelline, naringin, kaempferol, embinin, catechin, isorhamnetin, quercetin
(i) Suppressed proliferation(ii) Induced apoptosis independent from p53 stemness property (in HCCSCs)(iii) Antiproliferative properties
(i) Suppressed cell proliferation and expression of c-Myc and cyclin D1(ii) Induced intrinsic mitochondrial apoptotic signaling pathway
[140]
Red grape
NM
HT-29, HCT116
0.9-2.0 mg/mL
Hydroethanolic
Delphinidin glycosides, quercetin derivatives, delphinidin-3-O-glucoside (high), cyanidin-3-O-glucoside
(i) Highest growth inhibition(ii) Increased the percentage of apoptotic cells
(i) Downregulation of apoptotic proteins, such as cIAP-2, livin, survivin, and XIAP(ii) Inhibition of tyrosine kinase
[130]
Black lentil
NM
HT-29, HCT116
0.9-2.0 mg/mL
Hydroethanolic
Delphinidin glycosides, procyanidin B1, delphinidin-3-O-glucoside (high), cyanidin-3-O-glucoside
(i) Significantly arrested HT-29 cells in G1(ii) Highest growth inhibition(iii) Increased percentage of apoptotic cells
(i) Downregulation of apoptotic proteins, such as cIAP-2, livin, survivin, and XIAP(ii) Inhibition of tyrosine kinase
[130]
Graptopetalum paraguayense
Leaf
Caco-2, BV-2
0.2, 0.4, 0.6, 0.8, 1.0 mg/mL
Hydroethanolic
Oxalic acid, hydroxybutanedioic acid, gallic acid, quercetin, chlorogenic acid glucans with fucose, xylose, ribose (GW100) arabino-rhamnogalactans (GW100E)
(i) Great potential in antiproliferation(ii) Significant immunomodulatory activities on BV-2 cells and interleukin-6 (IL-6) (GW100)
(i) Scavengingα, α-diphenyl-β-picrylhydrazyl radicals (DPPH) (GW100E excelled in scavenging DPPH), 2,2-azino-bis [3-ethylbenzothiazoline-6-sulfonic acid] radicals (ABTS), superoxide anions (O2) (GW100)(ii) Significant inhibition of tumor necrosis factor-a (TNF-a), scavenging ABTS and O2
[141]
Butea monosperma
Flower
SW480
200, 370μg/mL
Floral
n-Butanol
Significant antiproliferative effect
(i) Significantly downregulated the expression of Wnt signaling proteins such asβ-catenin, APC, GSK-3β, cyclin D1, and c-Myc(ii) Increased intracellular level of ROS
[142]
Rehmannia glutinosa
NM
CT26
5, 20, 80μM
NM
Catalpol
Inhibited proliferation and growth invasion of colon cancer cells
(i) Downregulated MMP-2 and MMP-9 protein expressions(ii) Reduction in the angiogenic markers secretions
[143]
Telectadium dongnaiense
Bark
HCT116
1.5, 2.0μg/mL
MeOH extract
4-Dicaffeoylquinic acid, quercetin 3-rutinoside, periplocin
NM
(i) Inhibition ofβ-catenin/TCF transcriptional activity and effects on Wnt/β-catenin(ii) Downregulation of the expression of Wnt target genes
[144]
Gloriosa superba
Root
SW620
30 ng/mL
Protein hydrolysate extract
Protein hydrolysate
NM
(i) Upregulation of p53(ii) Downregulation of NFκB
[145]
Boswellia serrata
Resin
HT-29
100, 150μg
Methanolic
Boswellic acid
Decreased cell viability
(i) Reduction in mPGES-1, VEGF, CXCR4, MMP-2, MMP-9, HIF-1, PGE2 expression(ii) Increment in the caspase 3 activity(iii) Inhibition of cell migration and vascular sprout formation
[146]
Typhonium flagelliforme
Leaf
WiDr
70μg/mL
Ethyl acetate
Glycoside flavonoid, isovitexin, alkaloids
NM
Inhibition of COX-2 expression
[28]
Diospyros kaki
Fruit
HT-29
2,000μg/mL
Hydroacetone extract
Polyphenol
Impaired cell proliferation and invasion
NM
[147]
Carpobrotus edulis
Leaf
HCT116
1,000 mg/mL
Hydroethanolic
Gallic acid, quercetin, sinapic acid, ferulic acid, luteolin 7-o-glucoside, hyperoside, isoquercitrin, ellagic acid, isorhamnetin 3-O-rutinoside
Inhibited proliferation
(i) Possession of high DPPH scavenging activity and effective capacity for iron binding(ii) Inhibition of NO radical, linoleic acid peroxidation, protein glycation, and oxidative damage
[148]
Piper methysticum
Root
HT-29
10, 20, 30, 40, 50μg/mL
Aqueous
11-Hydroxy-12-methoxydihydrokavain, 11-hydroxy-12-methoxydihydrokavain, prenyl caffeate, pinostrobin chalcone, 11-methoxytetrahydroyangonin, awaine, methysticin, dihydromethysticin, 5,6,7,8-tetrahydroyangonin, kavain, 7,8-dihydrokavain, yangonin, desmethoxyyangonin, flavokawain B
Inhibited the growth
NM
[26]
Salvia ballotiflora
Ground aerial parts
CT26
6.76μg/mL
Hexane-washed chloroform extract
19-Deoxyicetexone, 7,20-dihydroanastomosine, icetexone, 19-deoxyisoicetexone
Cytotoxic activity
NM
[149]
Tinospora cordifolia
Stem
HCT116
1, 10, 30, 50μM
Hydroalcoholic
Clerodane furano diterpene glycoside, cordifoliosides A and Β, sitosterol, ecdysterone, 2β,3β:15,16-diepoxy-4α, 6β-dihydroxy-13(16),14-clerodadiene-17,12:18,1-diolide
Induced chromatin condensation and fragmentation of nuclei of few cells
(i) Considerable loss of MMP(ii) Decreased in mitochondria function(iii) Increased cytochrome c in the cytosol(iv) Induced ROS/oxidative stress(v) Increased autophagy
[150]
Euterpe oleracea
Fruit
NM
35μg/mL
Hydroethanolic
Vanillic acid, orientin, isoorientin
NM
(i) Scavenging capacity towards ROO and HOCl(ii) Inhibition of nitroso compound formation
[151]
Salvia miltiorrhiza
NM
HCT116
7.4±1.0, 4.4±0.5μg/mL
Ethanolic
Diterpene quinone
NM
Decreased levels of pro-caspases 3 and 9
[152]
Coffea
Bean
HCT116
1 mg/mL
Aqueous
Chlorogenic acid complex (CGA7)
NM
(i) DNA fragmentation, PARP-1 cleavage, caspase 9 activation, downregulation of Bcl-2 and upregulation of Bax
[153]
Illicium verum
Fruit
HCT116
10 mg/mL
Ethanolic
Gallic acid quercetin
Induction of apoptosis and inhibition of key steps of metastasis
NM
[154]
Garcinia propinqua Craib
Leaf
HCT116
NM
CH2Cl2 extract
Benzophenones, xanthones, and caged xanthones
Potent inhibitory cytotoxicities
NM
[155]
Stem, bark
HCT116
14.23, 23.95μM
MeOH, CH2Cl2, and EtOAc extract
Xerophenone A, doitunggarcinones A and B, sampsonione, 7β-H-11-benzoyl-5α-ydroxy-6, 10-tetramethyl-1-(3-methyl-2-butenyl)-tetracyclotetradecane-2,12,14-trione, hypersampsone M, assiguxanthone A (cudraxanthone Q), 40 10-O-methylmacluraxanthone (16), 41- and 5-O-methylxanthone V1
NM
NM
[156]
Malus pumila Miller cv. Annurca
Fruit
Caco-2
400 mg/L
Methanolic
Chlorogenic acid, (+)catechin, (–)epicatechin, isoquercetin, rutin, phloridzin, procyanidin B2, phloretin, quercetin
WNT inhibitors and reduced WNT activity elicited by WNT5A
NM
[157]
Malus domestica cv. Limoncella
Fruit
Caco-2
400 mg/L
Methanolic
Chlorogenic acid, (+)catechin, (–)epicatechin, isoquercetin, rutin, phloridzin, procyanidin B2, phloretin, quercetin
WNT inhibitors and reduced WNT activity elicited by WNT5A
NM
[157]
Coix lacryma-jobi var. ma-yuen
Leaf
HCT116
0.5, 1 mg/mL
Aqueous
Coixspirolactam A, coixspirolactam B, coixspirolactam C, coixlactam, methyl dioxindole-3-acetate
NM
Inhibited migration, invasion, and adhesion via repression of the ERK1/2 and Akt pathways under hypoxic conditions
[158]
Mesua ferrea
Stem, bark
HCT116, HT-29
3.3, 6.6, and 11.8μg/mL
NM
Fractions (α-amyrin, SF-3, n-Hex)
Downregulation of multiple tumor promoter
Upregulation of p53, Myc/Max, and TGF-β signaling pathways
[159]
Taraxacum
Root
SGC7901, BGC823
3 mg/mL
Aqueous
NM
NM
Proliferation and migration through targeting lncRNA-CCAT1
[160]
Portulaca oleracea
Leaf
HT-29 CSCs
2.25μg/mL
Alcoholic
Oxalic, malic acid
NM
Inhibited expression of the Notch1 andβ-catenin genes, regulatory and target genes that mediate the Notch signal transduction pathway
[161]
Hordeum vulgare L.
NM
HT-29
NM
Aqueous & juice
Protein, dietary fiber, the B vitamins, niacin, vitamin B6, manganese, phosphorus, carbohydrates
(i) Inhibited proliferation of cancer cells(ii) Cytotoxic activity
Free radical scavenging activity
[162]
Paraconiothyrium sp.
NM
COLO 205 and KM12
12.5μM
Methyl ethyl ketone extract
n-Hexane, CH2Cl2, EtOAc, and MeOH fractions (A−D)
(i) Growth inhibitory activity(ii) Antiproliferative effect
NM
[163]
Mentha×piperita
Leaf
HCT116
5, 10, 20, 30, 40, 50μg/mL
Aqueous
Polyphenols
NM
Inhibited replication of DNA and transcription of RNA which induce the ROS
[164]
Mammea longifolia Planch. and Triana
Fruit
SW480
25, 50, 100μg/mL
Methanolic
NM
NM
Mitochondria-related apoptosis and activation of p53
[165]
Rollinia mucosa (Jacq.) Baill.
NM
HCT116, SW-480
<4, <20μg/mL
EtOH
Rollitacin, jimenezin, membranacin, desacetyluvaricin, laherradurin
Cytotoxic activity
NM
[54]
Annona diversifolia Saff.
NM
SW-480
0.5μg/mL
NM
Cherimolin-2
Cytotoxic activity
NM
[54]
A. purpurea Moc. & Sessé ex Dunal
NM
HT-29
1.47μg/mL
CHCl3-MeOH
Purpurediolin, purpurenin, annoglaucin, annonacin A
Cytotoxic activity
NM
[54]
Viguiera decurrens (A.Gray) A. Gray
NM
NM
3.6μg/mL
Hex; EtOAc; MeOH
β-Sitosterol-3-O-β-D-glucopyranoside; β-D-glucopyranosyl oleanolate; β-sitosterol-3-O-β-D-glucopyranoside, and oleanolic acid-3-O-methyl-β-D-glucuronopyranoside ronoate
Cytotoxic activity
NM
[54]
Helianthella quinquenervis (Hook.) A. Gray
NM
HT-29
2-10μg/mL
NM
Demethylencecalin
Cytotoxic activity
NM
[54]
Smallanthus maculatus (Cav.) H. Rob.
NM
HCT15
<20μg/mL
Acetone
Fraction F-4, fraction F-5, ursolic acid
Cytotoxic activity
NM
[54]
Bursera fagaroides (Kunth) Engl.
NM
HF6
1.8×10-4 to 2.80 μg/mL
Hydroalcoholic
Podophyllotoxin,β-peltatin-A methyl ether, 5′-desmethoxy-β-peltatin-A methyl ether, desmethoxy-yatein, deoxypodophyllotoxin, burseranin, acetyl podophyllotoxin
NM
(i) Inhibitor of microtubules(ii) Ability to arrest cell cycle in metaphase
[54]
Viburnum jucundum C.V. Morton
NM
HCT15
<20μg/mL
Acetone
Ursolic acid
Cytotoxic activity
NM
[54]
Hemiangium excelsum (Kunth) A.C.Sm.
NM
HCT15
<10 (μg/mL)
MeOH
PE, EtOAc, MeOH
Cytotoxic activity
NM
[54]
Hyptis pectinata (L.) Poit.
NM
Col2
<4, <20μg/mL
NM
Pectinolide A, pectinolide B, pectinolide C,α-pyrone, boronolide, deacetylepiol-guine
Cytotoxic activity
NM
[54]
H. verticillata Jacq.
NM
Col2
<4,<20μg/mL
NM
Dehydro-β-peltatin, methyl ether dibenzylbutyrolactone, (-)-yatein, 4′-demethyl-deoxypodophyllotoxin
Nonspecific cytotoxic activity
NM
[54]
H. suaveolens (L.)
NM
HF6
2.8-12μg/mL
Chloroform and butanol
β-Apopicropodophyllin
Nonspecific cytotoxic activity
NM
[54]
Salvia leucantha Cav.
Leaf, root, stem
HF6, HT-29, HCT15
14.9, 12.7, 9.9μg/mL
CHCl3
NM
Cytotoxic activity
NM
[54]
Vitex trifolia L.
NM
HCT15
3.5 to <1 (μg/mL)
Hexane and dichloromethane
Salvileucalin B, Hex: leaf, Hex: stem, DCM: leaf, DCM: stem
Cytotoxic activity
NM
[54]
Persea americana Mill.
NM
HT-29
<4μg/mL and <20 μg/mL
Ethanolic
1,2,4-trihydroxynonadecan, 1,2,4-trihydroxyheptadec-16-ene, 1,2,4-trihydroxyheptadec-16-yne
Cytotoxic activity
NM
[54]
Linum scabrellum
Roots, aerial parts
HF6
0.2, 0.5, 2.3μg/mL
Chloroform and butanol
DCM: MeOH, 6MPTOXPTOX
NM
(i) Induction of cell cycle arrest in G2/M(ii) Inhibition of tubulin polymerization
[54]
Phoradendron reichenbachianum (Seem.) Oliv.
NM
HCT15
3.6, 3.9, and 4.3μg/mL
NM
Moronic acid
Cytotoxic activity
NM
[54]
Cuphea aequipetala Cav.
NM
HCT15
18.70μg/mL
Acetone
NM
Cytotoxic inactivity
NM
[54]
Galphimia glauca Cav.
NM
HCT15
0.63, 0.50, 1.99μg/mL
EtOH, MeOH, aqueous
NM
Cytotoxic activity
NM
[54]
Mimulus glabratus Kunth
NM
HF6
12.64μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
Picramnia antidesma Sw.
NM
HCT15
0.6 to 4.5μM
NM
10-Epi-uveoside, uveoside, picramnioside E, picramnioside D
Cytotoxic activity
NM
[54]
Penstemon barbatus (Cav.) Roth
NM
HF6
15.19μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
P. campanulatus (Cav.) Willd.
NM
HF6
6.74μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
Veronica americana Schwein. ex Benth.
NM
HF6
0.169 and 1.46μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
Zea mays L.
NM
HCT116, SW-480, SW-620
NM
NM
13-Hydroxy-10-oxo-trans-11-octadecenoic acid
Cytotoxic activity
NM
[54]
Colubrina macrocarpa (Cav.) G. Don
NM
HCT15
10, 2.1, 9.1μg/mL
PE, EtOAc, MeOH
NM
Cytotoxic activity
NM
[54]
Coix lacryma-jobi
Seed, endosperm, and hull
HT-29
0.1–1,000μg/mL
Methanolic, hexane
Phytosterols (campesterol, stigmasterol, andβ-sitosterol), gamma-linolenic acid (GLA), arachidonic acid (AA), eicosapentaenoicacid (EPA) and docosahexaenoic acid (DHA), linoleic acid
NM
(i) Influence of signal transduction pathways that involve the membrane phospholipids(ii) Enhancement of ROS generation and decrease of cell antioxidant capacity
[166]
Abutilon indicum
Leaf
HT-29
210μg/mL
Aqueous
Flavonoids (4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl, 2-ethoxy-4-vinylphenol, N,N′-dimethylglycine, lup-20(29)-en-3-one, linolenin, 1-mono-, 9-hexadecanoic acid methyl ester, linolenic acid methyl ester), phenolic (amino acids, terpenoids, fatty acids, methyl palmitoleate)
NM
(i) Increase in the levels of reactive oxygen species and simultaneous reduction in cellular antioxidant, mitochondrial membrane loss, DNA damage, and G1/S phase cell cycle arrest
[167]
Galla rhois
NM
HCT116, HT-29
12.5, 25, 50, 100, 200μg/mL
Aqueous with steaming process
Gallotannins
Increased contents of gallic acid and ellagic acid
(i) Induced apoptosis through the activation of caspases 3, 8, 9(ii) Modulated activation of mitogen and protein kinases, p38, and c-Jun NH2-terminal kinase
[168]
Artemisia annua Linné
Powder
HCT116
20, 30, 40, 60, 80, 100μg/mL
Ethanolic
Phenolic compounds
Inhibited cell viability and increased LDH release
(i) PTEN/p53/PDK1/Akt signal pathways through PTEN/p53 induce apoptosis(ii) Increased apoptotic bodies, caspase 3 and 7 activation(iii) Regulated cytochrome c translocation to the cytoplasm and Bax translocation to the mitochondrial membrane
[169]
Nelumbo nucifera stamen
Powder
HCT116
100, 200, 400μg/mL
Ethanolic crude
NM
NM
(i) Increased the sub-G1 population, mRNA levels of caspases 3 and 8, levels of IκBα and caspase 9(ii) Modulated the Bcl-2 family mRNA expression(iii) Reduced the mRNA levels of NFκB
[170]
Corn silk
NM
LoVo, HCT116
1.25, 2.5, 5, 10, 20μg/mL
Aqueous
Proteins, polysaccharides, flavonoid, vitamins, tannins, alkaloids, mineral salts, steroids
NM
(i) Increase in the Bax, cytochrome c, caspases 3 and 9 levels
[171]
Lycium barbarum L.
Powder
HT-29
1, 2, 3, 4, 5μg/mL
NM
Neoxanthin, all-trans-β-cryptoxanthin, polysaccharides, carotenoids, flavonoids
NM
(i) Upregulation of p53 and p21 expression(ii) Downregulation of the CDK2, CDK1, cyclin A, and cyclin B expression(iii) Arrest in the G2/M phase of cell cycle
[172]
Chrysobalanus icaco L.
Freeze-dried fruit
HT-29
1, 2.5, 5, 10, 20μg/mL
Crude ethyl acetate
Delphinidin, cyanidin, petunidin, and peonidin
NM
(i) Increased intracellular ROS production(ii) Decreased TNF-α, IL-1β, IL-6, and NFκB1 expressions
[173]
Zanthoxylum piperitum De Candolle
Fruit
Caco-2, DLD-1
200μg/mL
Aqueous
NM
NM
(i) Increased the phosphorylation of c-Jun N-terminal kinase (JNK)
[174]
Celtis aetnensis (Tornab.) Strobl (Ulmaceae)
Twigs
Caco-2
5, 50, 100, 250, or 500μg/mL
Methanolic
Flavonoid and triterpenic compounds
NM
(i) Increase in the levels of ROS(ii) Decrease in RSH levels and expression of HO-1
[175]
Rosa canina
Peel and pulp
Caco-2
62.5, 125, 250, 500μg/mL
Total extract (fraction 1), vitamin C (fraction 2), neutral polyphenols (fraction 3), and acidic polyphenols (fraction 4)
Polyphenols
Decreased production of reactive oxygen species (ROS)
NM
[176]
Rhazya stricta
Leaf
HCT116
47, 63, 79, and 95μg/cm2
Crude alkaloid
Alkaloids
NM
(i) Downregulated DNA-binding and transcriptional activities of NFκB and AP-1 proteins(ii) Increase in Bax, caspases 3/7 and 9, p53, p21 and Nrf-2 levels(iii) Decrease in expression of ERK MAPK, Bcl-2, cyclin D1, CDK-4, survivin, and VEGF
[177]
Green coffee
NM
Caco-2
10-1,000μg/mL
NM
5-Caffeoylquinic acid (5-CQA), 3,5-dicaffeoylquinic acid (3,5-DCQA), ferulic acid (FA), caffeic acid (CA), dihydrocaffeic acid (DHCA), dihydroferulic acid (DHFA)
Reduced viability of cancer cells
NM
[178]
Flourensia microphylla
Leaf
HT-29
NM
Ethanolic and acetone
Phenolic compounds
NM
(i) Inhibition of IL-8(ii) Activation of apoptosis by the increment of the Bax/Bcl-2 ratio and expression of TNF family
[179]
∗NM: not mentioned.


### 3.1.2. Studies in Animal Models

The most used animal model is the murine one (Tables2(a) and 2(b)). In particular, studies were carried out above all on HT-29 and HCT116 cells. The effects of the different medicinal plants and their extracts are essentially the same detected in in vitro studies. In particular, plant extracts were able to induce apoptosis and inhibit proliferation and tumor angiogenesis by regulating p53 levels and checkpoint proteins with consequent cell cycle arrest and antiproliferative and antiapoptotic effects on cancerous cells.Table 2
(a)
Efficacy of medicinal plants on colon cancer inin vivo models
Scientific name
Parts used
Model
Dose
Type of extract
Important compounds
Cellular effect
Mechanisms
References
Vitis vinifera
Seed
In vivo (murine)
Caco-2
In vivo: 400–1,000 mg/kgIn vitro: 10–25 μg/mL
Aqueous
Procyanidins
(i) Increased crypt depth and growth-inhibitory effects(ii) Inhibited cell viability(iii) Significantly decreased the histological damage score
Reduced MPO (myeloperoxidase) activity
[180]
Seed
In vivo
HT-29, SW480
5 mg/kg
Aqueous
NM
NM
Decreased VEGF, TNF, MMP-1, MMP-3, MMP-7, MMP-8, MMP-9, and MMP-13 protein expression
[181]
Skin
In vivo
NM
7.5, 30, 60μg/mL
Methanolic
4′-Geranyloxyferulic acid
NM
NM
[30]
Seed
In vivo (murine)
NM
0.12%w/w
NM
Catechin, epicatechin
NM
(i) Suppressed proliferation, sphere formation, nuclear translocation ofβ-catenin and Wnt/β-catenin signaling(ii) Elevated p53, Bax/Bcl-2 ratio, and cleaved PARP and mitochondrial-mediated apoptosis
[31]
Camellia sinensis
Leaf
In vivo (murine)
HT-29
In vitro: 0, 10, 30, 50μMIn vivo: 1.5 mg per day
Aqueous
Catechin, epigallocatechin gallate
1.9-fold increase in tumor endothelial cell apoptosis
Inhibited the ERK-1 and ERK-2 activation, VEGF expression, and VEGF promoter
[182]
In vivo (murine)
HCT116
0.5%
NM
NM
Reduced basement membrane
Inhibition of MMP-9 and VEGF secretion
[183]
In vivo (murine)
Caco-2, HT-29
300μM
Aqueous
Theaflavins (TF-2, TF-3, TF-1)
Induced apoptosis of human colon cancer cells
Inhibition of edema formation correlated to attenuation of COX-2 expression and promoter analysis revealed modulation of NFκB, AP-1, CREB, and/or NF-Il-6 (C/EBP)
[36]
In vivo (murine)
HT115
25μg/mL
Hydroethanolic
Phenolic compounds (p-hydroxyphenyl ethanol, pinoresinol & dihydroxyphenyl ethanol)
NM
Inhibition via reduced expression of a range ofα5 & β1
[184]
Sasa quelpaertensis
Leaf
In vivo
HT-29, HCT116
0, 100, 200, 300 mg/L
Ethanolic
p-Coumaric acid, tricin
Inhibition of colony formation
(i) Nonadherent sphere formation suppressed CD133+ & CD44+ population(ii) Downregulated expression of cancer stem cell markers
[41]
Anoectochilus
NM
In vivo
CT26
Oral dose of 50 & 10 mg/mouse per day
Aqueous
Kinsenoside
Stimulated proliferation of lymphoid tissues
Activation of phagocytosis of peritoneal macrophages
[185]
Purple-fleshed potatoes
Fruit
In vivo
Colon cancer stem cells
5.0μg/mL
Ethanol, methanol, ethyl acetate
Anthocyanin,β-catenin, cytochrome c
Reduction in colon CSCs number and tumor incidence
(i) Increase in cytochrome c levels from p53 status and maybe mitochondria-mediated apoptosis(ii) Suppressed levels of cytoplasmic and nuclear β-catenin
[58]
Phaseolus vulgaris
Leaf
In vivo
HT-29
Nm
Ethanolic
Polysaccharides, oligosaccharides
Induction of apoptosis and inhibit proliferation
(i) Inactivation of the retinoblastoma phosphoprotein(ii) Induced G1 arrest(iii) Suppression of NF-jb1(iv) Increase in EGR1 expression
[59]
Rosmarinus officinalis L.
Leaf
In vivo
HT-29
SC-RE 30μg/mL and CA 12.5 μg/mL
Ethanolic
Polyphenols (carnosic acid (CA) and carnosol)
(i) Activation of Nrf2 transcription factor(ii) Activated common regulators, such as XBP1 (Xbp1) gene, SREBF1/SREBF2 (Srebp1/2), CEBPA and NR1I2 (Pxr) genes
Leaf
In vivo (rat)
NM
NM
Ethanolic
Rosmanol and its isomers, carnosol, rosmadial, carnosic acid, and 12-methoxycarnosic acid, carnosic acid, carnosol
Interactions with the gut microbiota and by a direct effect on colonocytes with respect to the onset of cancer or its progression
NM
Wasabia japonica
Rhizomes
In vivo
COLO 205
5 mg/mL
Methanolic
6-(Methylsulfinyl)hexyl isothiocyanate
Anticolon cancer properties through the induction of apoptosis and autophagy
(i) Activation of TNF-α, Fas-L, caspases(ii) Truncated Bid and cytochrome c(iii) Decreased phosphorylation of Akt and Mtor(iv) Promoted expression of microtubule-associated protein 1 light chain 3-II and AVO formation
[186]
Zingiberaceae
Rhizome
HT-29
HT-29
5 g/kg
Dichloromethanic
Turmerone
Suppressed the proliferation of HT-29 colon cancer cells
(i) LDH release(ii) ROS generation(iii) Collapse in mitochondrial membrane potential(iv) Cytochrome c leakage(v) Activation of caspase 9 and caspase 3
[187]
Panax quinquefolius
Root
In vivo (murine)
NM
30 mg/kg
Ethanolic
Ginsenosides (protopanaxadiol or protopanaxatriol)
Attenuated azoxymethane/DSS-induced colon carcinogenesis by reducing the colon tumor number and tumor load
(i) Reduced experimental colitis(ii) Attenuated on AOM/DSS-induced colon carcinogenesis(iii) Proinflammatory cytokines activation(iv) Suppressed DSS(v) Downregulated inflammatory cytokine gene expression
[188]
Myrtaceae
Leaf
In vivo (murine)
HCT116
100μg/mL (in vitro) 200 and 100 μg/disc (in vivo)
Methanolic
Phenolics, flavonoids, betulinic acid
Inhibition of tumor angiogenesis
(i) Inhibition of angiogenesis of tube formation on Matrigel matrix and HUVECS migration (in vitro)(ii) Decreased nutrient and oxygen supply and consequently tumor growth and tumor size (in vivo)(iii) Increased extent of tumor necrosis
[82]
Spica prunellae
Leaf
In vivo
HT-29
200 mg/mL (in vitro), 600 mg/mL (in vivo)
Ethanolic
Rosmarinic acid
Induction of apoptosis and inhibition of cell proliferation and tumor angiogenesis
(i) Induced apoptosis(ii) Inhibited cancer cell proliferation and angiogenesis STAT3 phosphorylation(iii) Regulated expression of Bcl-2, Bax, cyclin D1, CDK4, VEGF-A, and VEGFR-2 (in vivo)
[83]
Gymnaster koraiensis
Aerial part
In vivo (murine)
NM
500μmol/kg
Ethanolic
Gymnasterkoreaynes B, C, E, 2,9,16-heptadecatrien-4,6-dyne-8-ol
Anti-inflammatory and cancer preventive activities
(i) Significant decrease in expression of COX-2(ii) Increase in serum IL-6
[189]
Allium fistulosum
Edible portions
In vivo (murine)
CT26
50 mg/kg b.w.
Hot water
p-Coumaric acid, ferulic acid, sinapic acid, quercitrin, isoquercitrin, quercetol, kaempferol
Suppression of tumor growth and enhanced survival rate of test mice
(i) Decreased expression of inflammatory molecular markers(ii) Downregulated expression of MMP-9 and ICAM(iii) Metabolite profiling and candidate active phytochemical components
[190]
Annona squamosa Linn
Leaf
In vivo (animal)
HCT116
8.98μg/mL
Crude ethyl acetate
Acetogenins (annoreticuin & isoannoreticuin) and alkaloids dopamine, salsolinol, and coclaurine
(i) Inhibited growth and proliferation of tumor cells
Reactive oxygen species (ROS) formation, lactate dehydrogenase (LDH) release, and caspases 3/7, 8, 9 activation
[191]
Eupatorium cannabinum
Aerial parts
In vivo (murine)
HT-29
25μg/mL
Ethanolic
Pyrrolizidine alkaloids (senecionine, senkirkine, monocrotaline, echimidine)
Cytotoxicity against colon cancer cells
(i) Upregulation of p21 and downregulation of NCL, FOS, and AURKA, indicating reduced proliferation capacity(ii) Mitotic disruption and nonapoptotic cell death via upregulation of Bcl-xL
[96]
Flacourtia indica
Aerial parts
In vivo (murine)
HCT116
500μg/mL
Methanolic
Phenolic glucoside (flacourticin, 4′-benzoylpoliothrysoside)
Antiproliferative and proapoptotic effects in HCT116 cells
Apoptosis via generation of ROS and activation of caspases (PARP)
[192]
Sorghum bicolor
The dermal layer of stalk
In vivo (murine)
HCT116 & colon cancer stem cells
>16 and 103μg/mL
Phenolic, acetone
Apigeninidin & luteolinidin
Antiproliferative effect
(i) Target p53-dependent and p53-independent pathways
[97]
Gleditsia sinensis
Thorn
In vivo (murine)
HCT116
800μg/mL
Aqueous
Flavonoid, lupine acid, ellagic acid glycosides
Inhibited proliferation of colon cancer
(i) Increased p53 levels(ii) Downregulation of the checkpoint proteins, cyclin B1, Cdc2, and Cdc25c
[90]
Thorn
In vivo (murine)
HCT116
600μg/mL
Ethanolic
NM
Inhibitory effect on the proliferation of human colon cancer HCT116 cells
(i) Caused G2/M phase cell cycle arrest
[91]
Zingiber officinale
Rhizome
In vitro/in vivo (murine)
HCT116
5μM
Ethanolic
6-Paradol, 6- and 10-dehydrogingerdione, 6- and 10-gingerdione, 4-, 6-, 8-, and 10-gingerdiol, 6-methylgingerdiol, zingerone, 6-hydroxyshogaol, 6-, 8-, 10-dehydroshogaol, diarylheptanoids
Inhibitory effects on the proliferation of human colon cancer cells
(i) Arrest of G0/G1 phase(ii) Reduced DNA synthesis(iii) Induced apoptosis
[103]
Cucumaria frondosa
The enzymatically hydrolyzed epithelium of the edible
In vivo (murine)
HCT116
<150μg/mL
Hydroalcoholic
Monosulphated triterpenoid glycoside frondoside A, the disulphated glycoside frondoside B, the trisulphated glycoside frondoside C
(i) Inhibition at S and G2-M phase with a decrease in Cdc25c(ii) Increase in p21WAF1/CIP
(i) Inhibition the growth of human colon(ii) Apoptosis associated with H2AX phosphorylation and caspase 2
[105]
Rolandra fruticosa
Leaf & twigs
In vivo (murine)
HT-29
10 and 5 mg/kg/day
Methanolic
Sesquiterpene lactone (13-acetoxyrolandrolide)
Antiproliferative effect against human colon cancer cells
(i) Inhibition of the NFκB pathway, subunit p65 (RelA) and upstream mediators IKKβ and oncogenic K-ras
[106]
Cydonia oblonga Miller
Leaf & fruit
In vivo (murine)
Caco-2
250–500μg/mL
Methanolic
Phenolic compound (flavonol and flavone heterosides, 5-O-caffeoylquinic acid)
Antiproliferative effect against human kidney and colon cancer cells
(i) Suppression of NFκB activation, activator (AP-1), mitogen-activated protein kinases, namely, PKC, (GFR)-mediated pathways(ii) Cell cycle arrest(iii) Induction of apoptosis, antioxidant, and anti-inflammatory effects
[107]
Sedum kamtschaticum
Aerial part
In vivo (murine)
HT-29
0–0.5 mg/mL
Methanolic
Buddlejasaponin IV
Induced apoptosis in HT-29 human colon cancer cells
(i) Induced apoptosis via mitochondrial-dependent pathway triggered by downregulation of Bcl-2 protein levels, caspase 3 activation, and subsequent PARP cleavage
[109]
Ganoderma lucidum
Caps & stalks
In vivo (murine)
HT-29
0-0.1 mg/mL
Triterpene extract (hot water) extract
Polysaccharides (mainly glucans & glycoproteins), triterpenes (ganoderic acids, ganoderic alcohols, and their derivatives)
Cytokine expression inhibited during early inflammation in colorectal carcinoma
Induced autophagy through inhibition of p38 mitogen-activated kinase and activation of farnesyl protein transferase (FPT)
[193]
Ginkgo biloba
Fruit & leaf
In vivo (murine)
HT-29
20–320 mg/L
Aqueous
Terpene lactones and flavonoid glycosides
Inhibited progression of human colon cancer cells induced HT-29 cell apoptosis
(i) Activation in caspase 3, reduction in Bcl-2 expression, and elevation in p53 expression
[112]
Rubus occidentalis
Fruit
In vivo (murine)
JB6 Cl 41
25μg/mL
Methanolic
β-Carotene, α-carotene, ellagic acid, ferulic acid, coumaric acid
Inhibited tumor development
(i) Impaired signal transduction pathways leading to activation of AP-1 and NFB RU-ME fraction
[194]
Oryza sativa
Seed
In vivo (murine)
HT-29, SW 480, HCEC
100μg/mL
Ethyl acetate extract
Phenolic compound (tricin, ferulic acid, caffeic acid, and methoxycinnamic acid)
Inhibited growth of human colon cancer cells
(i) Induction of apoptosis by enhanced activation of caspases 8 and 3(ii) Decreased the number of viable SW480 and HCEC cells
[113]
Cistanche deserticola
Dried stem
In vivo (murine)
SW480
In vivo: 0.4 g/kg/dayIn vitro: 100 mg/mL
Aqueous
Polysaccharides, phenylethanoid glycosides
Decreased mucosal hyperplasia and helicobacter infection
(i) Increased number of splenic macrophages and NK cells(ii) Decreased frequency of hyperplasia and H. hepaticus infection of the intestine
[133]
Rehmannia glutinosa
NM
In vivo (male C57BL6 mice and Sprague-Dawley rats)
CT26
28 mg/kg
NM
Catalpol
(i) Inhibited proliferation, growth, and expression of angiogenic markers
(i) VEGF, VEGFR2, HIF-1α, bFGF inhibited the expressions of inflammatory factors such as IL-1β, IL-6, and IL-8
[143]
Olea europaea
Olive mill wastewater
In vivo (murine)
NM
NM
Methanolic
Hydroxytyrosol
Interferes with tumor cell growth
NM
[195]
Leaf
In vivo (xenograft model) (murine)
HCT116, HCT8
0, 5, 10, 20, 30, 50, and 70μg/mL
Phenolic
Oleuropein and hydroxytyrosol
NM
(i) Activation of caspases 3, 7, and 9(ii) Decrease of mitochondrial membrane potential and cytochrome c release(iii) Increase in intracellular Ca2+ concentration
[196]
Ginkgo biloba L.
Leaf
In vivo (rat)
NM
0.675 and 1.35 g/kg
Methanolic
Flavonoid glycosides, terpene lactones, and ginkgolic acids
(i) Suppressed tumor cell proliferation, promoted apoptosis, and mitigated inflammation
NM
[197]
Rhus trilobata Nutt.
NM
In vivo (hamster)
NM
400 mg/kg, 100 mg/kg
Aqueous
Tannic acid, gallic acid
Cytotoxic activity
NM
[54]
Annona diversifolia Saff.
NM
In vivo (mice)
SW-480
1.5, 7.5 mg/kg/day
NM
Laherradurin
Cytotoxic activity
NM
[54]
A. muricata L.
NM
In vivo (rat)
NM
250/500 mg/kg
EtOAc
A, B, and C, and cis- and trans-annomuricin-D-ones
Cytotoxic activity
NM
[54]
Plumeria acutifolia Poir.
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Lasianthaea podocephala (A. Gray) K. M. Becker
NM
In vivo (hamster)
NM
200 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Flourensia cernua DC.
NM
In vivo (hamster)
NM
350 mg/kg/day
Aqueous
Flavonoids, sesquiterpenoids, monoterpenoids, acetylenes, p-acetophenones, benzopyrans, benzofurans
Cytotoxic activity
NM
[54]
Ambrosia ambrosioides (Cav.) W. W. Payne
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Alnus jorullensis Kunth
NM
In vivo (hamster)
NM
175 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Dimorphocarpa wislizeni (Engelm.) Rollins
NM
In vivo (hamster)
NM
100 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Euphorbia pulcherrima Willd. ex Klotzsch
NM
In vivo (hamster)
NM
200 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Acalypha monostachya Cav.
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Crotalaria longirostrata Hook. & Arn.
NM
In vivo (hamster)
NM
400 mg/kg/day, 350 mg/kg/day
EtOH-CHCl3
NM
Cytotoxic activity
NM
[54]
Asterohyptis stellulata (Benth.) Epling
NM
In vivo (hamster)
NM
50 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Acacia constricta A. Gray
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Holodiscus dumosus A. Heller
NM
In vivo (hamster)
NM
350 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Butea monosperma
Flower
In vivo (rat)
HT-29
150 mg/kg
n-Butanol extract
Isocoreopsin, butrin, and isobutrin
Free radical scavenging and anticancer activities
NM
[198]
Taraxacum spp.
Root
In vivo (xenograft murine model)
HT-29, HCT116
40 mg/kg/day
Aqueous
α-Amyrin, β-amyrin, lupeol, and taraxasterol
Induced programmed cell death
NM
[199]
∗NM: not mentioned.
(b)
Other effects of medicinal plants inin vivo models
Scientific name
Parts used
Model
Dose
Type of extract
Important compounds
Cellular effect
Mechanisms
References
Allium sativum
Root
In vivo (murine)
NM
2.4 mL of daily
Ethanolic
Allicin, S-allylmercaptocysteine
Significantly suppressed both the size and number of colon adenomas
Enhancement of detoxifying enzymes: SAC and GST activity
[200]
Olea europaea
Fruit
In vivo
Caco-2
50μM
Aqueous
Phenolic compounds, authentic hydroxyl tyrosol (HT)
(i) Effect of OPE and HT on CB1 associated with reduced proliferation of Caco-2 cells(ii) Increase in CB1 expression in the colon of rats receiving dietary EVOO
Increase in Cnr1 gene expression, CB1 protein levels
[201]
In vivo (murine)
HT115
25μg/mL
Hydroethanolic
Phenolic compounds (p-hydroxyphenyl ethanol, pinoresinol & dihydroxyphenyl ethanol)
NM
Inhibition via reduced expression of a range ofα5 & β1
[184]
Origanum vulgare L.
Leaf
In vivo (murine)
NM
20, 40, 60 mg·kg−1
Aqueous
Rosmarinic acid, caffeic acid, flavonoids
Antioxidant status
(i) Increased LPO products and activity of SOD and CAT enzymes and GST and GPx activity(ii) Antioxidant and anticarcinogenic effect
[202]
Hazelnut
Skin
In vivo
NM
The flow rate 0.21 mL/min and injection volume 9.4μL
Aqueous
Flavan-3-ols, in monomeric and polymeric forms, and phenolic acids
(i) Decreased circulating levels of free fatty acids and triglycerides(ii) Higher excretion of bile acid
Increase of the total antioxidant capacity of plasma
[203]
Apples and apple juice
Fruit
In vivo
NM
90 mg/L
Aqueous
Phenolic acids, flavonoids, tannins, stilbenes, curcuminoids
NM
NM
[204]
Grifola frondosa
Fruit
In vivo (murine)
HT-29
10 ng/mL
Aqueous
Phenolic compounds (pyrogallol, caffeic acid, myricetin, protocatechuic acid, etc.)
Inhibition of TNBS-induced rat colitis
(i) Induced cell cycle progression in G0/G1 phase and apoptotic death
[104]
Ruta chalepensis
Leaf
In vivo (human)
NM
250μg/mL
Ethanolic
Rutin, gallic acid, catechin hydrate, naringin
Oxidative profile in patients with colon cancer
NM
[205]
Cannabis sativa
Dry flower & leaf
In vivo (murine)
DLD-1 and HCT116
0.3–5μM
Methanolic
Cannabidiol, phytocannabinoids
NM
(i) Reduced cell proliferation in a CB1-sensitive and AOM-induced preneoplastic lesions and polyps(ii) Inhibition of colorectal cancer cell proliferation via CB1and CB2 receptor activation
[121]
Melia toosendan
Fruit
In vivo (murine)
SW480, CT26
0, 10, 20, 30, 40, 50μg/mL
Ethanolic
Triterpenoids, flavonoids, polysaccharide, limonoids
NM
(i) Inhibited cell proliferation of SW480 and CT26 by promoting apoptosis as indicated by nuclear chromatin condensation and DNA fragmentation(ii) Induced caspase 9 activity which further activated caspase 3 and poly(ADP-ribose) polymerase cleavage, leading the tumor cells to apoptosis
[123]
Smallanthus sonchifolius
Root
In vivo (murine)
NM
73.90, 150.74, 147.65, and 123.26 mg/kg
Aqueous
Fructans
NM
Reduction incidence of colon tumors expressing alteredβ-catenin
[206]
Punica granatum
Peel
In vivo (adult male Wistar rats)
NM
4.5 g/kg
Methanolic
Gallic acid, protocatechuic acid, cateachin, rutin, ellagic acid, punicalagin
NM
(i) Reduction in TGF-β, Bcl-2, EGF, CEA, CCSA-4, MMP-7 and in COX-2, cyclin D1, survivin content(ii) Downregulated expression of β-catenin, K-ras, c-Myc genes
[207]
Linum usitatissimum
Seed
In vivo (male Sprague-Dawley rats)
NM
500 mg/kg
Alkaline
Secoisolariciresinol diglucoside, carbohydrates, proteins, and tannins
Reduced the serum fasting glucose levels
Significantly reduced the HbA1c, insulin levels, and proinflammatory cytokines
[208]
Diospyros kaki
Fruit
In vivo (male CD-1 mice)
NM
15 mg/kg
Hydroacetone
Polyphenol
(i) Decreased attenuation of colon length in diarrhea severity(ii) Reduced mortality rate(iii) Reduction of the extent of visible injury (ulcer formation) and of mucosal hemorrhage
Decreased expression of COX-2 and iNOS in the colonic tissue
[147]
Muntingia calabura
Leaf
In vivo (rat)
NM
50, 250, 500 mg/kg
Methanolic
Rutin, gallic acid, ferulic acid, and pinocembrin
Reduction of the colonic oxidative stress, increasing the antioxidants levels possibly via the synergistic action of several flavonoids
NM
[209]
Portulaca oleracea
NM
In vivo (murine)
HT-29 CSCs
2.25μg/mL
Alcoholic
NM
Regulatory and target genes that mediate the Notch signal transduction pathway
Inhibition of expression of the Notch1 andβ-catenin genes
[161]
Aloe vera
Gel
In vivo (murine)
NM
400 mg/kg/day
Gel
Polysaccharides
NM
(i) Via inhibition of the cell cycle progression(ii) Induction of cellular factors, such as extracellular signal-regulated kinases 1/2, cyclin-dependent kinase 4, and cyclin D1; on the other hand, PAG increased the expression of caudal-related homeobox transcription factor 2
[210]
Artemisia annua Linné
Powder
In vivo (xenograft murine model)
HCT116
20, 40 mg/kg/day
Ethanolic
Phenolic compounds
NM
(i) Induced apoptosis via PTEN/p53/PDK1/Akt signal pathways through PTEN/p53(ii) Inhibited cell viability and increased LDH release and apoptotic bodies, caspase 3 and 7 activation, and reduced mitochondria membrane potential(iii) Regulated cytochrome c translocation to the cytoplasm and Bax translocation to the mitochondrial membrane(iv) Regulation of proteins
[169]
Hordeum vulgare
Powder
In vivo (xenograft murine model)
HT-29
2 g/kg and 1 g/kg
Aqueous (fermented)
β-Glucan, protein, amino acids, phenolic compounds
NM
(i) Promoted tumor apoptosis by upregulating the mRNA expression of Bax and caspase 3 and downregulating the mRNA expression of Bcl-2 and cyclin D1(ii) Decreased mRNA expression of Bcl-2 and cyclin D1(iii) Upregulated expressions levels of Bax and caspase 3
[211]
Dendrophthoe pentandra
Leaf
In vivo (murine)
NM
125, 250, 500 mg/kg
Ethanolic
Quercetin-3-rhamnose
NM
(i) Decreased the levels of IL-22, MPO levels, proliferation of epithelial cells(ii) Inhibited S phase of the cell cycle(iii) Upregulated p53 wild-type gene expression
[212]
Aquilaria crassna
Stem, bark
In vivo (murine)
HCT116
2,000 mg/kg/day100, 200 mg/kg
NM
Resin and essential oils
NM
NM
[213]
Berberis integerrima
NM
In vivo (murine)
NM
50 and 100 mg/kg
Hydroalcoholic
NM
NM
NM
[214]
Salix aegyptiaca
Bark
In vivo (murine)
NM
100 and 400 mg/kg
Ethanolic
Catechin, catechol, and salicin
NM
Decreased level of EGFR, nuclearβ-catenin, and COX-2
[215]The main mechanisms of action of medicinal plants are summarized in Figure1.Figure 1
Cell damage and cancer trigger p53 activation. The p53 protein activates the apoptotic protein Bax. Bax inhibits the antiapoptotic protein Bcl-2. During apoptosis, cytochrome c is released from mitochondria. To activate the Apaf-1 protein, the interaction between these proteins and cytochrome C is necessary. Pro-caspase 9 attaches to Apaf-1 and activates caspase 9. Caspase 9 activates caspases 3 and 7 and apoptosis occurs.Inin vitro studies, it has been found that grapes, which contain substantial amounts of flavonoids and procyanidins, play a role in reducing the proliferation of cancer cells by increasing dihydroceramides and p53 and p21 (cell cycle gate keeper) protein levels. Additionally, grape extracts triggered antioxidant response by activating the transcriptional factor nuclear factor erythroid 2-related factor 2 (Nrf2) [27].Grape seeds contain polyphenolic and procyanidin compounds, and their reducing effects on the activity of myeloperoxidase have been shown inin vitro and in vivo studies. It has been suggested that grape seeds could inhibit the growth of colon cancer cells by altering the cell cycle, which would lead eventually to exert the caspase-dependent apoptosis [180].Another plant that attracted researchers’ attention was soybean, which contain saponins. After 72 h of exposure of colon cancer cells to the soy extract, it was found that this extract inhibited the activity and expression of protein kinase C and cyclooxygenase-2 (COX-2) [34]. The density of the cancer cells being exposed to the soy extract significantly decreased. Soybeans can also reduce the number of cancer cells and increase their mortality, which may be due to increased levels of Rab6 protein [216].Green tea leaves have also attracted the researchers’ attention in these studies. Green tea leaves, with high levels of catechins, increased apoptosis in colon cancer cells and reduced the expression of the vascular endothelial growth factor (VEGF) and its promoter activity inin vitro and in vivo studies. The extract increased apoptosis (programmed cell death) by 1.9 times in tumor cells and 3 times in endothelial cells compared to the control group [182]. In another in vitro study, the results showed that green tea leaves can be effective in the inhibition of matrix metalloproteinase 9 (MMP-9) and in inhibiting the secretion of VEGF [183].Garlic was another effective plant in this study. Its roots have allicin and organosulfur compounds. In anin vitro study, they inhibited cancer cell growth and induced apoptosis through the inhibition of the phosphoinositide 3-kinase/Akt pathway. They can also increase the expression of phosphatase and tensin homolog (PTEN) and reduce the expression of Akt and p-Akt [32]. Garlic roots contain S-allylcysteine and S-allylmercaptocysteine, which are known to exhibit anticancer properties. The results of a clinical trial on 51 patients, whose illness was diagnosed as colon cancer through colonoscopy, and who ranged in age from 40 to 79 years, suggest that the garlic extract has an inhibitory effect on the size and number of cancer cells. Possible mechanisms suggested for the anticancer effects of the garlic extract are both the increase of detoxifying enzyme soluble adenylyl cyclase (SAC) and an increased activity of glutathione S-transferase (GST). The results suggest that the garlic extract stimulates mouse spleen cells, causes the secretion of cytokines, such as interleukin-2 (IL2), tumor necrosis factor-α (TNF-α), and interferon-γ, and increases the activity of natural killer (NK) cells and phagocytic peritoneal macrophages [200].The results ofin vitro studies on olive fruit showed that it can increase peroxide anions in the mitochondria of HT-29 cancer cells due to the presence of 73.25% of maslinic acid and 25.75% of oleanolic acid. It also increases caspase 3-like activity up to 6 times and induces programmed cell death through the internal pathway [217]. Furthermore, the olive extract induces the production of reactive oxygen species (ROS) and causes a quick release of cytochrome c from mitochondria to cytosol.The pomegranate fruit contains numerous phytochemicals, such as punicalagins, ellagitannins, ellagic acid, and other flavonoids, including quercetin, kaempferol, and luteolin glycosides. The results of anin vitro study indicate the anticancer activity of this extract through reduction of phosphorylation of the p65 subunit and subsequent inhibition of nuclear factor-κB (NFκB). It also inhibits the activity of TNF receptor induced by Akt, which is needed for the activity of NFκB. The fruit juice can considerably inhibit the expression of TNF-α-inducing proteins (Tipα) in the COX-2 pathway in cancer cells [43]. The effective and important compounds in pomegranate identified in these 104 studies are flavonoids, polyphenol compounds, such as caffeic acid, catechins, saponins, polysaccharides, triterpenoids, alkaloids, glycosides, and phenols, such as quercetin and luteolin, and kaempferol and luteolin glycosides.In a systematic review of the plants being studied, some mechanisms were mainly common, including the induction of apoptosis by means of an increase of expression and levels of caspase 2, caspase 3, caspase 7, caspase 8, and caspase 9 in cancer cells, increasing the expression of the proapoptotic protein Bax and decreasing the expression of the antiapoptotic proteins.Many herbal extracts block specific phase of the cell cycle. For instance, the extract prepared from the leaves ofAnnona muricata inhibits the proliferation of colon cancer cells and induces apoptosis by arresting cells in the G1 phase [53]. They can also prevent the progress of the G1/S phase in cancer cells [74]. In general, the herbal extracts reported here have been able to stop cancer cells at various stages, such as G2/M, G1/S, S phase, G0/G1, and G1 phase, and could prevent their proliferation and growth.Other important anticancer mechanisms are the increase of both p53 protein levels and transcription of its gene. Even the increase of p21 expression is not without effect [137]. In an in vitro study on the Garcinia mangostana roots, the results were indicative of the inhibitory effect of the extract of this plant on p50 and P65 activation [93]. Moreover, reduction of cyclin D1 levels and increase of p21 levels are among these mechanisms [137], as well as inhibition of NFκB and reduction of the transcription of its genes, which contribute to reduce the number of cancerous cells [127]. Other important anticancer mechanisms are the inhibition of COX-2, as well as the reduction of the protein levels in this pathway [34]. In addition to this, in some cases, the inhibition of MMP-9 can be mentioned as the significant mechanism of some herbal extracts to kill cancer cells [183].


## 3.1.1.In Vitro Studies

Out of 172 studies, 75 were carried out on HT-29, 60 on HCT116, and 24 on Caco-2 cells (Table1). On HT-29 cells, both Allium sativum root extracts and Camellia sinensis leaf extracts induced cell apoptosis by two different mechanisms, respectively. In fact, the former showed inhibition of the PI3K/Akt pathway, upregulation of PTEN, and downregulation of Akt and p-Akt expression, while the latter was involved in attenuation of COX-2 expression and modulation of NFκB, AP-1, CREB, and/or NF-IL-6. Moreover, an antiproliferative activity has also been detected in Olea europaea fruit extracts, which increased caspase 3-like activity and were involved in the production of superoxide anions in mitochondria. An antiproliferative activity, by means of a blockage in the G2/M phase, has also been reported in Caco-2 cells by Vitis vinifera fruit extracts. Concerning HCT116 cells, several plants, such as American ginseng and Hibiscus cannabinus, induced cell cycle arrest in different checkpoints.Table 1
Cytotoxic effects of medicinal plants on colon cancer inin vitro models.
Scientific name
Parts used
Cell line
Conc.
Type of extract
Important compounds
Cellular effect
Mechanisms
References
Vitis vinifera
Fruit
HCT116
NM
Lyophilized
Hydroxycinnamic acids, proanthocyanidins, stilbenoids
Increase of dihydroceramides, sphingolipid mediators involved in cell cycle arrest, and reduction of the proliferation rate
(i) Increase of p53 and p21 cell cycle gate keepers(ii) Activation of the transcriptional factor Nrf2
[26, 27]
Fruit
Caco-2
365 mg/g
Methanolic
Catechin, epicatechin, quercetin, gallic acid
Antiproliferative activity and direct initiation of cell death
Blockage in the G2/M phase
[28, 29]
Seed
Caco-2
10–25μg/mL
Aqueous
Procyanidins
(i) Increased crypt depth(ii) Inhibited cell viability and decreased histological damage score
Reduced MPO (myeloperoxidase) activity
[29]
Skin
NM
7.5, 30, 60μg/mL
Methanolic
4′-Geranyloxyferulic acid
NM
NM
[30]
Seed
Colon cancer stem cells
6.25, 12.5, 25μg/mL
NM
(+)-catechin, (−)-epicatechin
NM
(i) Increment of p53, Bax/Bcl-2 ratio, and cleaved PARP(ii) Inhibition of Wnt/β-catenin signaling
[31]
Allium sativum
Root
HT-29
20, 50, 100 mg/mL
Ethanolic
NM
Induction of apoptosis and cell cycle arrest
(i) Inhibition of the PI3K̸Akt pathway(ii) Upregulation of PTEN and downregulation of Akt and p-Akt expression
[32]
Glycine max
Seed
Caco-2, SW620, HT-29
12.5μg/mL
Aqueous
Anthoxanthin
Cell death and significant reduction of cell density
Enhancement of Rab6 protein levels
[33]
Seed
HT-29
240, 600 ppm
Crude
Saponin
Suppression of PKC activation and increase of alkaline phosphatase activity
[33]
Seed
HT-29
NM
Crude
Saponin
NM
(i) Suppression of the degradation of IκBα in PMA-stimulated cells(ii) Downregulation of COX-2 and PKC expressions
[34]
Camellia sinensis
Leaf
HT-29
0, 10, 30, 50μM
Aqueous
Catechin, epigallocatechin gallate
1.9-fold increase in tumor cell apoptosis and a 3-fold increase in endothelial cell apoptosis
(i) Suppression of ERK-1 and ERK-2 activation(ii) Suppression of VEGF expression
[35]
Leaf
Caco-2, HT-29
300μM
Aqueous
Theaflavins (TF-2T, F-3, TF-1)
Human colon cancer cell apoptosis induction
Modulation of NFκB, AP-1, CREB, and/or IL-6
[36]
Leaf
HT-29
68-800.73 μg/mL
Hot water extract
Flavan-3-ol (catechin & tannin) & polyphenols (teadenol B)
Inhibition of proliferation of HT-29 cells
Increased expression levels of caspases 3/7, 8, and 9
[35]
Olea europaea
Fruit
HT-29
150, 55.5200 and 74 μmol/L
Methanolic and chloroform
Maslinic acid, oleanolic acid
Antiproliferative activity
(i) Increased caspase 3-like activity to 6-fold(ii) Production of superoxide anions in the mitochondria
[37]
Fruit, leaf
SW480 and HT-29
100–400 m/z
Methanolic & hexane
Oleic acid, linoleic acid, gamma-linolenic acid, lignans, flavonoids, secoiridoids
Reduced cell growth in both cell lines
(i) Limited G2M cell cycle(ii) Depressed cyclooxygenase-2 expression in HT-29 cells(iii) Suppression of β-catenin/TCF signaling in SW480 cells(iv) Promotion of the entry into subG1 phase
[38]
Fruit
Caco-2
50μM
Aqueous
Phenolic compounds, authentic hydroxyl tyrosol (HT)
Reduced proliferation of Caco-2 cells
Reduction of the methylation levels of CNR1 promoter
[39]
Fruit
HT115
25μg/mL
Hydroethanolic
Phenolic compounds (p-hydroxyphenyl ethanol, pinoresinol & dihydroxyphenyl ethanol)
NM
Inhibition by reduced expression of a range ofα5 & β1
[40]
Olive mill wastewater
HT-29, HCT116, CT26
NM
Methanolic
Hydroxytyrosol
(i) Inhibited proliferation(ii) Inhibited migration and invasion
(i) Reduced sprout formation(ii) Inhibited VEGF and IL-8 levels
[41]
Fruit
Caco-2
0-2,000μg/mL
Ethanolic
Tyrosol, hydroxytyrosol, oleuropein, rutin, quercetin and glucoside forms of luteolin and apigenin
NM
(i) Induction of the cell cycle arrest in S-phase
[42]
Punica granatum
Juice
HT-29
50 mg/L
Aqueous
Ellagitannins, punicalagin
Inhibition of cancer cell proliferation
(i) Suppressed TNFR-induced COX-2 protein expression(ii) Reduced phosphorylation of the p65 subunit and binding to the NFκB response element
[43]
Seed
LS174
63.2μg/mL
Supercritical fluid
Punicic acid,γ-tocopherol, α-tocopherol
Cytotoxic activity
(i) Slightly decreased development of tubules from elongated cell bodies(ii) Reduction of the number of cell connections
[44]
Glycyrrhiza glabra
Root
HT-29
12.2 and 31μg/mL
Ethanolic
Licochalcone
NM
Increase of the protein levels of proapoptotic Bax
[37]
Opuntia ficus-indica
Fruit
Caco-2
115μM
Aqueous
Betalain pigment indicaxanthin
Apoptosis of proliferating cells
(i) Demethylation of the tumor suppressor p16INK4a gene promoter(ii) Reactivation of the silenced mRNA expression and accumulation of p16INK4a
[38]
Fruit
HT-29 & Caco-2 & NIH 3 T3 (as control)
Against HT-29 (4.9μg/mL) against Caco-2 (8.2 μg/mL)
Alkaline hydrolysis with NaOH
Isorhamnetin glycosides (IG5 and IG6)-phenol
Cell death through apoptosis and necrosis
Increased activity of caspase 3/7
[45]
Piper betle
Leaf
HT-29 and HCT116
200.0μg/mL
Aqueous
Hydroxychavicol
Antioxidant capacity and induction of a greater apoptotic effect
(i) Scavenging activity(ii) Formation of electrophilic metabolites
[41, 46]
Fragaria×ananassa
Fruit
HT-29
0.025, 0.05, 0.25, 0.5%
Ethanolic
Ascorbate, ellagic acid
Decreased proliferation of HT-29 cells
Increase in the levels of 8OHA and decrease in the levels of 8OHG
[40]
Sasa quelpaertensis
Leaf
HT-29 HCT116
0, 100, 200, 300 mg/L
Ethanolic
p-Coumaric acid, tricin
Inhibited colony formation
Nonadherent sphere formation suppressed CD133+ & CD44+ population
[41]
Salvia chinensis
Stem
HCT116, COLO 205
10, 20, 40,60, 80, 100 mg/L
Polyphenolic
Terpenoids, phenolic acid, flavonoids, dibenzylcyclooctadiene
Apoptosis & loss of mitochondrial membrane
Induced G0/G1 cell cycle
[42]
Rubus idaeus L.
Fruit
HT-29, HT-115, Caco-2
3.125, 6.25, 12.5, 25, 50 mg/L
Acetate
Polyphenol, anthocyanin, ellagitannin
NM
Decreased population of cells in G1 phase
[47]
LoVo
50μL
Aqueous
NM
Inhibited proliferation of LoVo
Suppression of the NFκB pathway
[48]
Curcuma longa
Root
HT-29, HCT15, DLD1, HCT116
(i) Short-term assay: four 10-fold dilutions (100 to 0.1 mg/L)(ii) Long-term assay: 5, 10, 20 mg/L
Ethanolic
Curcumin (diferuloylmethane)
Inhibited formation of HCT116 spheroids
NM
[49]
Eleutherococcus senticosus
Root
HCT116
12.5, 25, 50, 100
Methanolic
Eleutherosides, triterpenoid saponins, glycans
NM
Activation of natural killer cells and thus enhancement of immune function
[50]
Tabernaemontana divaricata L.
Leaf
HT-29, HCT15
10, 30, 100 mg/L
Ethyl acetate, chloroform, methanolic
Alkaloids
NM
Inhibited the unwinding of supercoiled DNA
[45]
Millingtonia hortensis
Root, flower, leaf
RKO
50, 100, 200, 400, 800 mg/L
Aqueous
Phenylethanoid glycoside, squalene, salidroside, 2-phenyl rutinoside
Apoptosis induction
(i) Increase of fragmented DNA(ii) Decrease of the expression of antiapoptotic proteins, Bcl-xL and p-BAD
[46]
Powder
RKO
200, 400, 800μg/mL
Aqueous
Water soluble compounds
Antiproliferative effect
NM
[51]
Thai purple rice
Seed
Caco-2, Cat. No. HTB-37
16.11μg/mL
Methanol acidified
Cyanidin-3-glucoside and peonidin-3-glucoside, anthocyanins, phenolic compounds
(i) Antioxidation of anthocyanins and phenols(ii) Antiproliferation of colon cancer cells
NM
[52]
Annona muricata
Leaf
HCT116, HT-29
11.43±1.87μg/ml and 8.98±1.24μg/ml
Ethanolic
Alkaloids, acetogenins, essential oils
Block of the migration and invasion of HT-29 and HCT116 cells
(i) Cell cycle arrest at G1 phase(ii) Disruption of MMP, cytochrome c leakage and activation
[53]
NM
HT-29, HCT116
<4, <20μg/mL
EtOAc
Annopentocin A, annopentocin B, annopentocin C, cis- and trans-annomuricin-D-ones, annomuricin E
NM
Suppression of ATP production and NADH oxidase in cancer cells
[54]
Pistacia lentiscus L. var. chia
Leaf
HCT116
NM
Ethanolic
Resin, known as Chios mastic gum (CMG)
Causes several morphological changes typical of apoptosis in cell organelles
(i) Induction of cell cycle arrest at G1 phase(ii) Activation of pro-caspases 8, 9, 3
[55]
Resin
HCT116
100μg/mL
Hexane
Caryophyllene
Induction of the anoikis form of apoptosis in human colon cancer HCT116 cells
(i) Induction of G1 phase arrest(ii) Loss of adhesion to the substrate
[56]
American ginseng (Panax quinquefolius)
Biological constituents
HCT116
0-2.0 mg/mL
Aqueous
Ginseng (GE) or its ginsenoside (GF) and polysaccharide (PS)
Proliferation was inhibited by GE, GF, and PS in wild-type and p21 cells
(i) Cells arrest in G0/G1 phase and increment of p53 and p21 proteins(ii) Increment of Bax and caspase 3 proteins expression
[57]
Purple-fleshed potatoes
Fruit
Colon cancer stem cells
5.0μg/mL
Ethanol, methanol, ethyl acetate
Anthocyanin,β-catenin, cytochrome c
Critical regulator of CSC proliferation and its downstream proteins (c-Myc and cyclin D1) and elevated Bax and cytochrome c
(i) Cytochrome c levels were elevated regardless of p53 status(ii) Mitochondria-mediated apoptotic pathway(iii) Suppressed levels of cytoplasmic and nuclear β-catenin
[58]
Phaseolus vulgaris
Leaf
HT-29
NM
Ethanolic
Polysaccharides, oligosaccharides
Changes in genes involved or linked to cell cycle arrest
(i) Inactivation of the retinoblastoma phosphoprotein(ii) Induction of G1 arrest(iii) Suppression of NF-jB1(iv) Increase in EGR1 expression
[59]
Opuntia spp.
Fruit
HT-29
5.8±1.0, 7.5±2.0, 12±1% (V/V)
Hydroalcoholic
Betacyanins, flavonoids (isorhamnetin derivatives) and phenolic acids (ferulic acid)
NM
Induced cell cycle arrest at different checkpoints—G1, G2/M, and S
[60]
Suillus luteus
NM
HCT15
400μg/mL
Methanolic
Protocatechuic acid, cinnamic acid,α-tocopherol, β-tocopherol, mannitol, trehalose, polyunsaturated fatty acids, monounsaturated fatty acids, saturated fatty acids
(i) Increase in the cellular levels of p-H2A.X, which is suggestive of DNA damage
(i) Inhibition of cell proliferation in G1 phase(ii) Increase in the cellular levels of p-H2A.X
[61]
Poncirus trifoliata
Leaf
HT-29
0.63μM
Aqueous (in acetone)
β-Sitosterol, 2-hydroxy-1,2,3-propanetricarboxylic acid 2-methyl ester
Arrest of cell growth was observed withβ-sitosterol
NM
[62]
Rosmarinus officinalis L.
Leaf
SW 620, DLD-1
0-120μg/mL
Methanolic
Polyphenols
Antiproliferative effect of 5-FU
Downregulation of TYMS and TK1, enzymes related to 5-FU resistance
[63]
Leaf
HT-29
SC-RE 30μg/mL and CA 12.5 μg/mL
Ethanolic
Polyphenols (carnosic acid (CA) and carnosol)
(i) Upregulation of VLDLR gene as the principal contributor to the observed cholesterol accumulation in SC-RE-treated cells(ii) Downregulation of several genes involved in G1-S
Activation of Nrf2 transcription factor and common regulators, such as XBP1 (Xbp1) gene related to the unfolded protein response (UPR)
[64]
NM
HT-29
10, 20, 30, 40, 50, 60, 70μg/mL
NM
Carnosic acid, carnosol, rosmarinic acid, rosmanol
NM
NM
[65]
Leaf
HGUE-C-1, HT-29, and SW480
20–40 mg/mL
CO2-supercritical fluid extract
Carnosic acid, carnosol, and betulinic acid
NM
(i) Prooxidative capability by increasing the intracellular generation of ROS(ii) Activation of Nrf2
[66]
Glehnia littoralis
Leaf
HT-29
50 mg/mL
Methanolic
Bergapten, isoimpinellin, xanthotoxin, imperatorin, panaxydiol, falcarindiol, falcarinol
Induced apoptosis by the decreased expression of the antiapoptotic Bcl-2 mRNA
(i) Reduced expression of Bcl-2(ii) Reduced expression levels of iNOS and COX-2
[67]
Verbena officinalis
Leaf
HCT116
20 mg/mL
Aqueous
Phenylethanoid glycosides, diacetyl-O-isoverbascoside, diacetyl-O-betonyoside A, and diacetyl-O-betonyoside A
(i) Substantial tumor cell growth inhibitory activity(ii) Time-dependent cytotoxicity against both cell lines
(i) Increased lipophilicity of molecules seemed to be responsible for enhanced cytotoxicity(ii) Antiproliferative activity is determined by the number of acetyl groups and also by their position in the aliphatic rings
[68]
Mentha spicata
Leaf
RCM-1
12.5μg/mL
N-Hexane
Acetic acid 3-methylthio propyl ester (AMTP), methyl thio propionic acid ethyl ester (MTPE)
Exhibited antimutagenic activity
Auraptene (7-geranyloxycoumarin) having a monoterpene moiety andβ-cryptoxanthin (one of the tetraterpenes) increased antibody production
[69]
Euphoria longana Lam.
Seed
SW 480
0–100μg/mL
Ethanolic
Corilagin, gallic acid, ellagic acid
(i) Antiangiogenetic properties(ii) All fractions showed the anti-VEGF secretion activity
Release and expression of VEGF indicated that all fractions showed the anti-VEGF secretion activity
[70]
Sutherlandia frutescens
Flower
Caco-2
1/50 dilution of the ethanolic extract
Ethanolic
Amino acids, including L-arginine and L-canavanine, pinitol, flavonoids, and triterpenoid saponins as well as hexadecanoic acid andγ-sitosterol
Disruption of the key molecules in the PI3K pathway thereby inducing apoptosis
Decrease in cell viability and increment in pyknosis as well as loss in cellular membrane integrity
[71]
Melissa officinalis
Leaf
HT-29, T84
346, 120μg/mL
Ethanolic
Phenolic acids (rosmarinic acid, coumaric acid, caffeic acid, protocatechuic acid, ferulic acid, chlorogenic acid), flavonoids, sesquiterpenes, monoterpenes, triterpenes
(i) Inhibited proliferation of colon carcinoma cells(ii) Induced apoptosis through formation of ROS
(i) G2/M cell cycle arrest(ii) Cleavage of caspases 3 and 7(iii) Induced phosphatidylserine externalization in colon carcinoma cells(iv) Induced formation of ROS in colon carcinoma cells
[72]
Sargassum cristaefolium
Leaf
HT-29
500 mg/mL
Ethanolic
Fucoidans
(i) Reduction of free radicals(ii) DPPH radical scavenging
Accumulation of cells in G0/G1 phase
[73]
Hedyotis diffusa
NM
HT-29
400 mg/mL
Ethanolic and then DMSO
Octadecyl (E)-p-coumarate, P-E-methoxy-cinnamic acid, ferulic acid, scopoletin, succinic acid, aurantiamide acetate, rubiadin
Suppress tumor cell growth and induce the apoptosis of human CRC cells
(i) Block G1/S progression(ii) Induce the activation of caspases 9 and 3(iii) Inhibit IL-6-mediated STAT3 activation(iv) Downregulate the mRNA and protein expression levels of cyclin D1, CDK4, Bcl-1, and Bax
[74]
Zingiber officinale Roscoe
Peel
LoVo
100 mg/mL
Ethanolic
Linoleic acid methyl ester,α-zingiberene, and zingiberone
Interesting antiproliferative activity against colorectal carcinoma
NM
[75]
Scutellaria barbata
Leaf
LoVo
413.3 mg/L
Methanolic
Scutellarein, scutellarin, carthamidin, isocarthamidin, wogonin
Induce cell death in the human colon cancer cell line
Increase in the sub-G1 phase and inhibition of cell growth
[76]
Pistacia atlantica, Pistacia lentiscus
Resin
HCT116
100μg/mL
Hexane extract
Caryophyllene
Induce the anoikis form of apoptosis in human colon cancer HCT116 cells
(i) Induce G1 phase arrest(ii) Loss of adhesion to the substrate
[56]
Citrus reticulata
Peel
SNU-C4
100μg/mL
Methanolic
Limonene, geranial, neral, geranyl acetate, geraniol,β-caryophyllene, nerol, neryl acetate
Induce the apoptosis on SNU-C4, human colon cancer cells
Expression of proapoptotic gene, Bax, and major apoptotic gene, caspase 3
[77]
Echinacea pallida, Echinacea angustifolia, Echinacea purpurea
Root
COLO320
150 mg/mL
Hexanic
Caffeic acid derivatives, alkylamides, polyacetylenes, polysaccharides
Induce apoptosis and promote nuclear DNA fragmentation
(i) Induce apoptosis by increasing caspase 3/7 activity(ii) Promote nuclear DNA fragmentation
[78]
Nasturtium officinale
Leaf
HT-29
50μL/mL
Methanolic
Phenethyl isothiocyanate, 7-methylsulfinylheptyl, 8-methylsulfinyl
(i) Inhibition of initiation, proliferation, and metastasis
(i) Inhibited DNA damage(ii) Accumulation of cells in S phase of the cell cycle
[79]
Polysiphonia
NM
SW480, HCT15, HCT116, DLD-1
20 and 40μg/mL
Methanolic
2,5-Dibromo-3,4-dihydroxybenzyl n-propyl ether
Potentially could be used as a chemopreventive agent against colon cancer
(i) Inhibited Wnt/β-catenin pathway(ii) Repressed CRT in colon cancer cells(iii) Downregulated cyclin D1(iv) Activated the NFκB pathway
[80]
Aristolochia debilis Sieb. et Zucc.
Stem
HT-29
200μg/mL
Methanolic
Aristolochic acid, nitrophenanthrene carboxylic acids
Inhibition of proliferation and induction of apoptosis in HT-29 cells
(i) Induction of sub-G1 cell cycle(ii) Generation of ROS and decrease of the MMP(iii) Bax overexpression and increase of Bax/Bcl-2 ratio
[81]
Myrtaceae
Leaf
HCT116
100μg/mL (in vitro), 200 and 100 μg/disc (in vivo)
Methanolic
Phenols, flavonoid, betulinic acid
Strong inhibition of microvessel outgrowth
(i) Inhibition of tube formation on Matrigel matrix(ii) Inhibition of HUVECS migration (in vitro)(iii) Decreased nutrient and oxygen supply
[82]
Spica prunellae
Leaf
HT-29
200 mg/mL (in vitro), 600 mg/mL (in vivo)
Ethanolic
Rosmarinic acid
Inhibits CRC cell growth
(i) Suppresses STAT3 phosphorylation(ii) Regulates the expression of Bcl-2, Bax, cyclin D1, CDK4, VEGF-A, and VEGFR-2
[83]
Phytolacca americana
Root
HCT116
3200μg/mL
Ethanolic
Jaligonic acids, kaempferol, quercetin, quercetin 3-glucoside, isoquercitrin, ferulic acid
Control of growth and spread of cancer cells
Reduction in the expressions of MYC, PLAU, and TEK
[84]
Morus alba
Leaf
HCT15
13.8μg/mL
Methanolic
Epicatechin, myricetin, quercetin hydrate, luteolin, kaempferol, ascorbic acid, gallic acid, pelargonidine, p-coumaric acid
Cytotoxic effect on human colon cancer cells (HCT15)
(i) Apoptosis induction also involved in the downregulation of iNOS(ii) Fragmentation of DNA(iii) Upregulation of caspase 3 activity
[85]
Rhodiola imbricata
Leaf
HT-29
200μg/mL
Acetone and methanolic
Phenols, tannins, and flavonoids
(i) Antioxidant activity(ii) Inhibited proliferation of HT-29 cells
(i) Scavenge free radicals(ii) DPPH radical scavenging activity(iii) Increased metal chelating activity
[86]
Asiasarum heterotropoides F.
DriedA. radix
HCT116
20 mg/mL
Ethanolic
Asarinin and xanthoxylol
Inhibition of the growth of HCT116 cells
(i) Caspase-dependent apoptosis(ii) Regulation of p53 expression at transcription level
[87]
Podocarpus elatus
Fruit
HT-29
500 mg/mL
Methanolic
Phenolic and anthocyanin
Reduction of proliferation of colon cancer cells
(i) Cell cycle delay in S phase(ii) 93% downregulation of telomerase activity and decrease in telomere length(iii) Induced morphological alterations to HT-29 cells
[88]
Echinacea purpurea
Flower
Caco-2, HCT116
0–2,000 mg/mL
Hydroethanolic
Cichoric acid
(i) Inhibition of proliferation(ii) Decreased telomerase activity in HCT116 cells
(i) Decreased telomerase activity(ii) Activation of caspase 9(iii) Cleavage of PARP(iv) Downregulation of β-catenin
[89]
Root
COLO320
150 mg/mL
Hexanic
Caffeic acid derivatives, alkylamides, polyacetylenes, polysaccharides
Induce apoptosis by increasing significantly caspase 3/7 activity and promote nuclear DNA fragmentation
(i) Increase significantly caspase 3/7 activity(ii) Promote nuclear DNA fragmentation
[78]
Hop (Humulus lupulus L.), Franseria artemisioides
Leaf
NM
100 mg/kg b.w./day
Aqueous
Coumarin, lignans, quinones
30% reduction of tumor-induced neovascularization
NM
[90]
NM
Caco-2
NM
Ethanolic
Phenolic compounds, flavonoid, diterpenes
Digestive, gastroprotective, antiseptic, anti-inflammatory, and antiproliferative activity
NM
[91]
Fruit
NL-17
0, 50, 100, 150μg/mL
Methanolic
α-Mangostin (xanthone)
NM
(i) Induction of caspase 3 and caspase 9 activation(ii) Induced cell cycle arrest at G1/G0 phase
[92]
Stem, bark
HT-29
50μg/mL
Chloroform-soluble
β-Mangostin, garcinone D, cratoxyxanthone
Cytotoxic activity against HT-29 human colon cancer
Inhibition of p50 and p65 activation
[93]
Annona squamosa Linn
Leaf
HCT116
8.98μg/mL
Crude, Aq ethyl acetate
Acetogenins (annoreticuin & isoannoreticuin) and alkaloids dopamine, salsolinol, and coclaurine
Inhibition of growth and proliferation of tumor cells
(i) Reactive oxygen species (ROS) formation, lactate dehydrogenase (LDH) release(ii) Activation of caspases 3/7, 8, and 9
[94]
Derris scandens
Stem
HT-29
5-15μg/mL
Ethanolic
Benzyls and isoflavones (genistein, coumarins, scandinone)
Apoptosis and mitotic catastrophe of human colon cancer HT-29 cells
(i) Inhibition ofα-glucosidase activity(ii) Scavenge free radicals
[95]
Eupatorium cannabinum
Aerial parts
HT-29
25μg/mL
Ethanolic
Pyrrolizidine alkaloids (senecionine, senkirkine, monocrotaline, echimidine)
Induced alteration of colony morphology
(i) Upregulation of p21 and downregulation of NCL, FOS, and AURKA(ii) Mitotic disruption and nonapoptotic cell death via upregulation of Bcl-xL, limited TUNEL labeling, and nuclear size increase
[96]
Sorghum bicolor
The dermal layer of stalk
HCT116 & colon cancer stem cells
>16 and 103μg/mL
Phenolic-rich ethanolic, acetone
Apigeninidin & luteolinidin
Antiproliferative
Target p53-dependent and p53-independent pathways
[97]
Dermal and seed head
CCSC
NM
Methanolic
Apigeninidin, luteolinidin, malvidin 3-O-glucoside, apigenin, luteolin, naringenin, naringenin 7-O-glucoside, eriodictyol 5-glucoside, taxifolin, catechins
NM
(i) Elevation of caspase 3/7 activity(ii) Decrease in β-catenin, cyclin D1, c-Myc, and survivin protein levels(iii) Suppression of Wnt/β-catenin signaling in a p53-dependent (dermal layer) and partial p53-dependent (seed head) manner
[98]
Hibiscus cannabinus
Seed
HCT116
KSE (15.625μg/mL to 1,000 μg/mL)
Ethanolic
Gallic acid, p-hydroxybenzoic acid, caffeic acid, vanillic acid, syringic acid, and p-coumaric and ferulic acids
Cytotoxic activity against human colon cancer HCT116 cells
Apoptosis via blockade of mid G1-late G1-S transition thereby causing G1 phase cell cycle arrest
[99]
Salix aegyptiaca L.
Bark
HCT116 & HT-29
300μg/mL
Ethanolic
Catechin, salicin, catechol and smaller amounts of gallic acid, epigallocatechin gallate (EGCG), quercetin, coumaric acid, rutin, syringic acid, and vanillin
Anticarcinogenic effects in colon cancer cells
Apoptosis via inhibition of phosphatidylinositol 3-kinase/protein kinase B and mitogen-activated protein kinase signaling pathways
[100]
Rubus coreanum
Fruit
HT-29
400μg/mL
Aqueous
Polyphenols, gallic acid, sanguine
Induction of apoptosis
(i) Induced activity of caspases 3, 7, and 9(ii) Cleavage of poly(adenosine diphosphate-ribose) polymerase
[101]
Codonopsis lanceolata
Root
HT-29
200μg/mL
N-Butanol fraction
Tannins, saponins, polyphenolics, alkaloids
Apoptosis in human colon tumor HT-29 cells
(i) Induced G0/G1 arrest(ii) Enhancement of expression of caspase 3 and p53 and of the Bax/Bcl-2 ratio
[102]
Gleditsia sinensis
Thorn
HCT116
800μg/mL
Aqueous
Flavonoid, lupine acid, ellagic acid glycosides
(i) Increase in p53 levels(ii) Downregulation of the checkpoint proteins, cyclin B1, Cdc2, and Cdc25c
Inhibition of proliferation of colon cancer cells
[90]
Thorn
HCT116
600μg/mL
Ethanolic
NM
Inhibitory effect on proliferation of human colon cancer HCT116 cells
(i) Caused cell cycle arrest at G2/M phase together with a decrease of cyclin B1 and Cdc2(ii) Progression from G2/M phase
[91]
Ligustrum lucidum
Fruit
DLD-1
50μg/mL
Aqueous
Oleanolic acid, ursolic acid
Inhibited proliferation
(i) Reduction of Tbx3 rescued the dysregulated P14ARF-P53 signaling
[94]
Zingiber officinale
Rhizome
HCT116
5μM
Ethanolic
6-Paradol, 6- and 10-dehydrogingerdione, 6- and 10-gingerdione, 4-, 6-, 8-, and 10-gingerdiol, 6-methylgingerdiol, zingerone, 6-hydroxyshogaol, 6-, 8-, 10-dehydroshogaol, diarylheptanoids
Inhibitory effects on the proliferation of human colon cancer cells
(i) Arrest at G0/G1 phase(ii) Reduced DNA synthesis
[103]
Grifola frondosa
Fruit
HT-29
10 ng/mL
Aqueous
Phenolic compounds (pyrogallol, caffeic acid, myricetin, protocatechuic acid)
Inhibition of TNBS-induced rat colitis
Induced cell cycle progression in G0/G1 phase
[104]
Cucumaria frondosa
The enzymatically hydrolyzed epithelium of the edible
HCT116
<150μg/mL
Hydroalcoholic
Monosulphated triterpenoid glycoside frondoside A, the disulphated glycoside frondoside B, the trisulphated glycoside frondoside C
Inhibition of human colon cancer cell growth
(i) Inhibition at S and G2-M phases with a decrease in Cdc25c and increase in p21WAF1/CIP(ii) Apoptosis associated with H2AX phosphorylation and caspase 2
[105]
Rolandra fruticosa
Leaf & twigs
HT-29
10 and 5 mg/kg/day
Methanolic
Sesquiterpene lactone (13-acetoxyrolandrolide)
Antiproliferative effect against human colon cancer cells
Inhibition of the NFκB pathway, NFκB subunit p65 (RelA), upstream mediators IKKβ and oncogenic K-ras
[106]
Cydonia oblonga Miller
Leaf & Fruit
Caco-2
250–500μg/mL
Methanolic
Phenolic compound (flavonol and flavone heterosides, 5-O-caffeoylquinic acid)
Antiproliferative effect against human kidney and colon cancer cells
(i) Suppression of factor activation, nuclear factor-kB (NFκB) activation, protein-1 (AP-1) transcription factor, mitogen protein kinases (MAPKs), protein kinases (PKs), namely, PKC, growth-factor receptor- (GFR-) mediated pathways and angiogenesis(ii) Cell cycle arrest and induction of apoptosis, antioxidant, and anti-inflammatory effects
[107]
Morchella esculenta
Fruits
HT-29
820 mg/mL
Methylene chloride
Steroids (mainly ergosterol derivatives) & polysaccharides & galactomannan
Antioxidant activity in HT-29 colon cancer cells
Inhibition of NF-B activation in the NF-B assay
[108]
Sedum kamtschaticum
Aerial part
HT-29
0–0.5 mg/mL
Methanolic
Buddlejasaponin IV
Induced apoptosis in HT-29 human colon cancer cells
Induction of apoptosis via mitochondrial pathway by downregulation of Bcl-2 protein levels, caspase 3 activation, and subsequent PARP cleavage
[109]
Ginseng and Glycyrrhiza glabra
Leaf
HT-29
500μL
Aqueous
Uracil, adenine, adenosine, Li-glycyrrhetinic acid, quiritin
NM
Antiproliferative effect determination of the protein levels of p21, cyclin D1, PCNA, and cdk-2, which are the key regulators for cell cycle progression
[110]
Orostachys japonicus
Leaf & stem
HT-29
2 mg/mL
Aqueous
Flavonoids, triterpenoids, 4-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, polysaccharide
Antiproliferation in HT-29 colon cancer cells
Inhibited proliferation at G2 point of the cell cycle and apoptosis via tumor suppressor protein p53
[111]
Ginkgo biloba
Fruit & leaf
HT-29
20–320mg/L
Aqueous
Terpene lactones and flavonoid glycosides
(i) Inhibited progression of human colon cancer cells(ii) Induced HT-29 cell apoptosis
Increase in caspase 3 activities and elevation in p53 MRN reduction in Bcl-2 mRNA
[112]
Oryza sativa
Seed
HT-29, SW 480, HCEC
100μg/mL
Ethyl acetate
Phenolic compound (tricin, ferulic acid, caffeic acid, and methoxycinnamic acid)
Inhibition of the human colon cancer cell growth
(i) Induced apoptosis by enhanced activation of caspases 8 and 3(ii) Decrease of the number of viable SW480 and HCEC cells(iii) Reduced colony-forming ability of these cells
[113]
Cnidium officinale Makino
Root
HT-29
305.024/mL
Ethanolic
Osthole, auraptenol, imperatorin
Inhibited proliferation of human colon cancer cells (HT-29)
Inhibition of the cellular proliferation via G0/G1 phase arrest of the cell cycle and induced apoptosis
[114]
Cnidium officinale Makino
Root
HT-29
0.1-5 mg/mL
Aqueous
N-(3-(Aminomethyl)benzyl)acetamidine
Inhibited the invasiveness of cytokine-treated HT-29 cells through the Matrigel-coated membrane in a concentration-dependent manner
(i) Reduction of HT-29 cell invasion through the Matrigel(ii) Inhibited cytokine-mediated NO production, iNOS expression, and invasiveness of HT-29 cells(iii) Inhibited MMP-2 activity
[115]
Long pepper (PLX)
Fruit
HT-29 and HCT116
0.10 mg/mL
Ethanolic
Piperidine alkaloids, piperamides, piperlongumine
(i) Induction of apoptosis, following DNA fragmentation in HT-29 colon cancer cells in a time-dependent manner(ii) Induced caspase-independent apoptosis
Induced whole cell ROS production
[116]
Achyranthes aspera
Root
COLO 205
50-100 and 150-200μg/mL
Ethanolic (EAA) and aqueous (AAA) root extractsAqueous
Phenolic compounds
(i) Enhanced growth inhibitory effects of AAA towards COLO 205 cells in contrast to EAA(ii) Stimulatory role of AAA in the activation of cell cycle inhibitors
(i) Triggered mitochondrial apoptosis pathway and S phase cell cycle arrest(ii) Increased levels of caspase 9, caspase 3, and caspase 3/7 activity
[117]
Thymus vulgaris
Leaf
HCT116
0.2, 0.4, 0.6, 0.8 mg/mL
Carvacrol and thymol
Inhibited proliferation, adhesion, migration, and invasion of cancer cells
[118]
Dictyopteris undulata
NM
SW480
40μg/mL
Ethanolic
Cyclozonarone benzoquinone
NM
Induced apoptosis by reducing Bcl-2 levels, upregulating Bax, and disrupting the mitochondrial membrane potential, leading to the activation of caspases 3 and 9
[119]
Dendrobium microspermae
NM
HCT116
0.25, 0.5, 1.0 mg/mL
Methanolic
NM
NM
Upregulation of Bax and caspases 9 and 3 and downregulation of Bcl-2 expression of genes
[120]
Cannabis sativa
Dry flower & leaf
DLD-1 and HCT116
0.3–5μM
Methanolic
Cannabidiol, phytocannabinoids
Reduced cell proliferation in a CB1-sensitive
(i) Reduced AOM-induced preneoplastic lesions and polyps(ii) Inhibited colorectal cancer cell proliferation via CB1 and CB2 receptor activation
[121]
Phoenix dactylifera L.
Fruit
Caco-2
0.2 mg/mL
Aqueous
Phenolic acids (gallic, protocatechuic, hydroxybenzoic, vanillic, isovanillic, syringic, caffeic, ferulic, sinapic, p-coumaric, isoferulic), flavonoid glycosides (quercetin, luteolin, apigenin, and kaempferol), and anthocyanidins
Increasing beneficial bacterial growth and inhibition of proliferation of colon cancer cells
NM
[122]
Melia toosendan
Fruit
SW480, CT26
0, 10, 20, 30, 40, 50μg/mL
Ethanolic
Triterpenoids, flavonoids, polysaccharide, limonoids
NM
(i) Inhibited cell proliferation of SW480 and CT26 by promoting apoptosis as indicated by nuclear chromatin condensation and DNA fragmentation(ii) Induced caspase 9 activity which further activated caspase 3 and poly(ADP-ribose) polymerase cleavage, leading the tumor cells to apoptosis
[123]
Crocus sativus L.
Flower
HCT116
0.25, 0.5, 1, 2, 4μg/mL
Ethanolic
Carotenoid, pigment, crocin, crocetin
Induced DNA damage and apoptosis
(i) Induction of a p53 pattern-dependent caspase 3 activation with a full G2/M stop(ii) Induced remarkable delay in S/G2 phase transit with entry into mitosis
[124]
Tepals and leaf
Caco-2
0.42 mg/mL
NM
Polyphenols, glycosides of kaempferol, luteolin, and quercetin
Proliferation of Caco-2 cells was greatly inhibited
NM
[125]
Luffa echinata
Fruit
HT-29
50, 100, and 200μg/mL
Methanolic
Amariin, echinatin, saponins, hentriacontane, gypsogenin, cucurbitacin B, datiscacin, 2-O-β-D-glucopyranosyl cucurbitacin B, and 2-O-β-D-glucopyranosyl cucurbitacin S
Increase in the population of apoptotic cells
(i) Inhibited the cellular proliferation of HT-29 cells via G2/M phase arrest of the cell cycle(ii) Induced apoptotic cell death via ROS generation(iii) Accumulation of caspase 3 transcripts of HT-29 cells
[126]
Vitis aestivalis hybrid
Fruits (wine)
CCD-18Co
25, 50, 100μg/mL
NM
Polyphenolics
NM
(i) Decreased mRNA expression of lipopolysaccharide- (LPS-) induced inflammatory mediators NFκB, ICAM-1, VCAM-1, and PECAM-1(ii) Enhanced expression of miR-126(iii) Decreased gene expression and reduced activation of the NFκB transcription factor, NFκB-dependent(iv) Decrease in ROS 113MAH
[127]
Xylopia aethiopica
Dried fruit
HCT116
0, 5, 10, 15, 20, 25, 30μg/mL
Ethanolic
Ent-15-oxokaur-16-en-19-oic acid (EOKA)
NM
(i) Induced DNA damage, cell cycle arrest in G1 phase, and apoptotic cell death
[128]
Sorghum
Grain
ER-β; nonmalignant young adult mouse colonocytes
1, 5, 10, 100μg/mL
Aqueous
Flavones (luteolin and apigenin), 3-deoxyanthocyanins naringenin (eriodictyol and naringenin)
Reduced cell growth via apoptosis
Increased caspase 3 activity
[129]
NM
HT-29, HCT116
0.9-2.0 mg/mL
Hydroethanolic
Procyanidin B1, delphinidin-3-O-glucoside, tannin, cyanidin-3-O glucoside
(i) Significantly arrested HT-29 cells in G1(ii) Highest growth inhibition(iii) Increased percentage of apoptotic cells
(i) Downregulation of apoptotic proteins, such as cIAP-2, livin, survivin, and XIAP, was seen in HCT116 cells(ii) Inhibition of tyrosine kinase
[130]
Panax notoginseng (Burk.) F.H. Chen
Root
LoVo and Caco-2
0, 100, 250, and 500μg/mL
Alcoholic
Saponin, ginsenoside
NM
Delay in progression of the G0/G1, S, or G2/M cell cycle phases
[131]
Brassica oleracea L. var. italica
Broccoli florets
HCT116
0, 1, 2.5, 5, 10μg/mL
Ethanolic
Glucoiberin, 3 hydroxy,4(α-L-rhamnopyranosyloxy), benzyl glucosinolate 4-vinyl-3-pyrazolidinone 4-(methyl sulphinyl), butyl thiourea, β-thioglucoside N-hydroxysulphates
NM
NM
[132]
Cistanche deserticola
Dried stem
SW480
In vivo: 0.4 g/kg/dayIn vitro: 100 μg/mL
Aqueous
Polysaccharides, phenylethanoid glycosides
(i) Decreased number of mucosal hyperplasia and intestinal helicobacter infection(ii) Increased number of splenic macrophage, NK cells, and splenic macrophages
Decreased frequency of hyperplasia andHelicobacter hepaticus infection of the intestine
[133]
Chaenomeles japonica
Fruit
Caco-2 and HT-29
10, 25, 50, 75, 100, 125, 150μM CE
NM
Procyanidins
NM
NM
[134]
Prunus mume
Fruit
SW480, COLO, and WiDr
150, 300, and 600μg/mL
Hydrophobic
Triterpenoid saponins
NM
(i) Inhibited growth and lysed SW480, COLO, and WiDr(ii) Induction of massive cytoplasmic vacuoles
[135]
Solanum lyratum
NM
COLO 205
50, 100, 200, 300, 400μg/mL
EtOH
β-Lycotetraosyl
Induced S phase arrest and apoptosis
(i) Induced DNA fragments(ii) Increased the levels of p27, p53, cyclin B1, active-caspase 3, and Bax(iii) Decreased the levels of Cdk1, pro-caspase 9, Bcl-2 and NF-ÎB, p65, and p50
[136]
Onopordum cynarocephalum
Aerial parts
HCT116, HT-29
0, 0.04, 0.12, 0.2, 0.4, 1.2 mg/mL0, 0.2, 0.4, 1.2, 2.0, 3.0 mg/mL
Aqueous
Flavonoids, lignans, and sesquiterpene lactones
NM
(i) Increase in the expression of proapoptotic proteins such as p53, p21, and Bax(ii) Inhibition of the antiapoptotic protein Bcl-2(iii) Decrease in cyclin D1 protein
[137]
Eleutherine palmifolia
Bulbs
SW480
2.5, 5, 10μg/mL
MeOH
Eleutherin, isoeleutherin
NM
(i) Inhibited the transcription of TCF/β-catenin(ii) Decrease in the level of nuclear β-catenin protein
[138]
Asparagus officinalis
Spears
HCT116
76μg/mL
Acetone
Steroidal saponins (HTSA-1, HTSAP-2, HTSAP-12, HTSAP-6, HTSAP-8)
NM
(i) Inhibition of Akt, p70S6K, and ERK phosphorylation(ii) Induction of caspase 3 activity, PARP-1 cleavage, DNA fragmentation, G0/G1 cell cycle arrest by reducing the expression of cyclins D, A, and E
[139]
Phyllanthus emblica L.
Seed, pulp
HCCSCs, HCT116
200μg/mL
Methanolic
Trigonelline, naringin, kaempferol, embinin, catechin, isorhamnetin, quercetin
(i) Suppressed proliferation(ii) Induced apoptosis independent from p53 stemness property (in HCCSCs)(iii) Antiproliferative properties
(i) Suppressed cell proliferation and expression of c-Myc and cyclin D1(ii) Induced intrinsic mitochondrial apoptotic signaling pathway
[140]
Red grape
NM
HT-29, HCT116
0.9-2.0 mg/mL
Hydroethanolic
Delphinidin glycosides, quercetin derivatives, delphinidin-3-O-glucoside (high), cyanidin-3-O-glucoside
(i) Highest growth inhibition(ii) Increased the percentage of apoptotic cells
(i) Downregulation of apoptotic proteins, such as cIAP-2, livin, survivin, and XIAP(ii) Inhibition of tyrosine kinase
[130]
Black lentil
NM
HT-29, HCT116
0.9-2.0 mg/mL
Hydroethanolic
Delphinidin glycosides, procyanidin B1, delphinidin-3-O-glucoside (high), cyanidin-3-O-glucoside
(i) Significantly arrested HT-29 cells in G1(ii) Highest growth inhibition(iii) Increased percentage of apoptotic cells
(i) Downregulation of apoptotic proteins, such as cIAP-2, livin, survivin, and XIAP(ii) Inhibition of tyrosine kinase
[130]
Graptopetalum paraguayense
Leaf
Caco-2, BV-2
0.2, 0.4, 0.6, 0.8, 1.0 mg/mL
Hydroethanolic
Oxalic acid, hydroxybutanedioic acid, gallic acid, quercetin, chlorogenic acid glucans with fucose, xylose, ribose (GW100) arabino-rhamnogalactans (GW100E)
(i) Great potential in antiproliferation(ii) Significant immunomodulatory activities on BV-2 cells and interleukin-6 (IL-6) (GW100)
(i) Scavengingα, α-diphenyl-β-picrylhydrazyl radicals (DPPH) (GW100E excelled in scavenging DPPH), 2,2-azino-bis [3-ethylbenzothiazoline-6-sulfonic acid] radicals (ABTS), superoxide anions (O2) (GW100)(ii) Significant inhibition of tumor necrosis factor-a (TNF-a), scavenging ABTS and O2
[141]
Butea monosperma
Flower
SW480
200, 370μg/mL
Floral
n-Butanol
Significant antiproliferative effect
(i) Significantly downregulated the expression of Wnt signaling proteins such asβ-catenin, APC, GSK-3β, cyclin D1, and c-Myc(ii) Increased intracellular level of ROS
[142]
Rehmannia glutinosa
NM
CT26
5, 20, 80μM
NM
Catalpol
Inhibited proliferation and growth invasion of colon cancer cells
(i) Downregulated MMP-2 and MMP-9 protein expressions(ii) Reduction in the angiogenic markers secretions
[143]
Telectadium dongnaiense
Bark
HCT116
1.5, 2.0μg/mL
MeOH extract
4-Dicaffeoylquinic acid, quercetin 3-rutinoside, periplocin
NM
(i) Inhibition ofβ-catenin/TCF transcriptional activity and effects on Wnt/β-catenin(ii) Downregulation of the expression of Wnt target genes
[144]
Gloriosa superba
Root
SW620
30 ng/mL
Protein hydrolysate extract
Protein hydrolysate
NM
(i) Upregulation of p53(ii) Downregulation of NFκB
[145]
Boswellia serrata
Resin
HT-29
100, 150μg
Methanolic
Boswellic acid
Decreased cell viability
(i) Reduction in mPGES-1, VEGF, CXCR4, MMP-2, MMP-9, HIF-1, PGE2 expression(ii) Increment in the caspase 3 activity(iii) Inhibition of cell migration and vascular sprout formation
[146]
Typhonium flagelliforme
Leaf
WiDr
70μg/mL
Ethyl acetate
Glycoside flavonoid, isovitexin, alkaloids
NM
Inhibition of COX-2 expression
[28]
Diospyros kaki
Fruit
HT-29
2,000μg/mL
Hydroacetone extract
Polyphenol
Impaired cell proliferation and invasion
NM
[147]
Carpobrotus edulis
Leaf
HCT116
1,000 mg/mL
Hydroethanolic
Gallic acid, quercetin, sinapic acid, ferulic acid, luteolin 7-o-glucoside, hyperoside, isoquercitrin, ellagic acid, isorhamnetin 3-O-rutinoside
Inhibited proliferation
(i) Possession of high DPPH scavenging activity and effective capacity for iron binding(ii) Inhibition of NO radical, linoleic acid peroxidation, protein glycation, and oxidative damage
[148]
Piper methysticum
Root
HT-29
10, 20, 30, 40, 50μg/mL
Aqueous
11-Hydroxy-12-methoxydihydrokavain, 11-hydroxy-12-methoxydihydrokavain, prenyl caffeate, pinostrobin chalcone, 11-methoxytetrahydroyangonin, awaine, methysticin, dihydromethysticin, 5,6,7,8-tetrahydroyangonin, kavain, 7,8-dihydrokavain, yangonin, desmethoxyyangonin, flavokawain B
Inhibited the growth
NM
[26]
Salvia ballotiflora
Ground aerial parts
CT26
6.76μg/mL
Hexane-washed chloroform extract
19-Deoxyicetexone, 7,20-dihydroanastomosine, icetexone, 19-deoxyisoicetexone
Cytotoxic activity
NM
[149]
Tinospora cordifolia
Stem
HCT116
1, 10, 30, 50μM
Hydroalcoholic
Clerodane furano diterpene glycoside, cordifoliosides A and Β, sitosterol, ecdysterone, 2β,3β:15,16-diepoxy-4α, 6β-dihydroxy-13(16),14-clerodadiene-17,12:18,1-diolide
Induced chromatin condensation and fragmentation of nuclei of few cells
(i) Considerable loss of MMP(ii) Decreased in mitochondria function(iii) Increased cytochrome c in the cytosol(iv) Induced ROS/oxidative stress(v) Increased autophagy
[150]
Euterpe oleracea
Fruit
NM
35μg/mL
Hydroethanolic
Vanillic acid, orientin, isoorientin
NM
(i) Scavenging capacity towards ROO and HOCl(ii) Inhibition of nitroso compound formation
[151]
Salvia miltiorrhiza
NM
HCT116
7.4±1.0, 4.4±0.5μg/mL
Ethanolic
Diterpene quinone
NM
Decreased levels of pro-caspases 3 and 9
[152]
Coffea
Bean
HCT116
1 mg/mL
Aqueous
Chlorogenic acid complex (CGA7)
NM
(i) DNA fragmentation, PARP-1 cleavage, caspase 9 activation, downregulation of Bcl-2 and upregulation of Bax
[153]
Illicium verum
Fruit
HCT116
10 mg/mL
Ethanolic
Gallic acid quercetin
Induction of apoptosis and inhibition of key steps of metastasis
NM
[154]
Garcinia propinqua Craib
Leaf
HCT116
NM
CH2Cl2 extract
Benzophenones, xanthones, and caged xanthones
Potent inhibitory cytotoxicities
NM
[155]
Stem, bark
HCT116
14.23, 23.95μM
MeOH, CH2Cl2, and EtOAc extract
Xerophenone A, doitunggarcinones A and B, sampsonione, 7β-H-11-benzoyl-5α-ydroxy-6, 10-tetramethyl-1-(3-methyl-2-butenyl)-tetracyclotetradecane-2,12,14-trione, hypersampsone M, assiguxanthone A (cudraxanthone Q), 40 10-O-methylmacluraxanthone (16), 41- and 5-O-methylxanthone V1
NM
NM
[156]
Malus pumila Miller cv. Annurca
Fruit
Caco-2
400 mg/L
Methanolic
Chlorogenic acid, (+)catechin, (–)epicatechin, isoquercetin, rutin, phloridzin, procyanidin B2, phloretin, quercetin
WNT inhibitors and reduced WNT activity elicited by WNT5A
NM
[157]
Malus domestica cv. Limoncella
Fruit
Caco-2
400 mg/L
Methanolic
Chlorogenic acid, (+)catechin, (–)epicatechin, isoquercetin, rutin, phloridzin, procyanidin B2, phloretin, quercetin
WNT inhibitors and reduced WNT activity elicited by WNT5A
NM
[157]
Coix lacryma-jobi var. ma-yuen
Leaf
HCT116
0.5, 1 mg/mL
Aqueous
Coixspirolactam A, coixspirolactam B, coixspirolactam C, coixlactam, methyl dioxindole-3-acetate
NM
Inhibited migration, invasion, and adhesion via repression of the ERK1/2 and Akt pathways under hypoxic conditions
[158]
Mesua ferrea
Stem, bark
HCT116, HT-29
3.3, 6.6, and 11.8μg/mL
NM
Fractions (α-amyrin, SF-3, n-Hex)
Downregulation of multiple tumor promoter
Upregulation of p53, Myc/Max, and TGF-β signaling pathways
[159]
Taraxacum
Root
SGC7901, BGC823
3 mg/mL
Aqueous
NM
NM
Proliferation and migration through targeting lncRNA-CCAT1
[160]
Portulaca oleracea
Leaf
HT-29 CSCs
2.25μg/mL
Alcoholic
Oxalic, malic acid
NM
Inhibited expression of the Notch1 andβ-catenin genes, regulatory and target genes that mediate the Notch signal transduction pathway
[161]
Hordeum vulgare L.
NM
HT-29
NM
Aqueous & juice
Protein, dietary fiber, the B vitamins, niacin, vitamin B6, manganese, phosphorus, carbohydrates
(i) Inhibited proliferation of cancer cells(ii) Cytotoxic activity
Free radical scavenging activity
[162]
Paraconiothyrium sp.
NM
COLO 205 and KM12
12.5μM
Methyl ethyl ketone extract
n-Hexane, CH2Cl2, EtOAc, and MeOH fractions (A−D)
(i) Growth inhibitory activity(ii) Antiproliferative effect
NM
[163]
Mentha×piperita
Leaf
HCT116
5, 10, 20, 30, 40, 50μg/mL
Aqueous
Polyphenols
NM
Inhibited replication of DNA and transcription of RNA which induce the ROS
[164]
Mammea longifolia Planch. and Triana
Fruit
SW480
25, 50, 100μg/mL
Methanolic
NM
NM
Mitochondria-related apoptosis and activation of p53
[165]
Rollinia mucosa (Jacq.) Baill.
NM
HCT116, SW-480
<4, <20μg/mL
EtOH
Rollitacin, jimenezin, membranacin, desacetyluvaricin, laherradurin
Cytotoxic activity
NM
[54]
Annona diversifolia Saff.
NM
SW-480
0.5μg/mL
NM
Cherimolin-2
Cytotoxic activity
NM
[54]
A. purpurea Moc. & Sessé ex Dunal
NM
HT-29
1.47μg/mL
CHCl3-MeOH
Purpurediolin, purpurenin, annoglaucin, annonacin A
Cytotoxic activity
NM
[54]
Viguiera decurrens (A.Gray) A. Gray
NM
NM
3.6μg/mL
Hex; EtOAc; MeOH
β-Sitosterol-3-O-β-D-glucopyranoside; β-D-glucopyranosyl oleanolate; β-sitosterol-3-O-β-D-glucopyranoside, and oleanolic acid-3-O-methyl-β-D-glucuronopyranoside ronoate
Cytotoxic activity
NM
[54]
Helianthella quinquenervis (Hook.) A. Gray
NM
HT-29
2-10μg/mL
NM
Demethylencecalin
Cytotoxic activity
NM
[54]
Smallanthus maculatus (Cav.) H. Rob.
NM
HCT15
<20μg/mL
Acetone
Fraction F-4, fraction F-5, ursolic acid
Cytotoxic activity
NM
[54]
Bursera fagaroides (Kunth) Engl.
NM
HF6
1.8×10-4 to 2.80 μg/mL
Hydroalcoholic
Podophyllotoxin,β-peltatin-A methyl ether, 5′-desmethoxy-β-peltatin-A methyl ether, desmethoxy-yatein, deoxypodophyllotoxin, burseranin, acetyl podophyllotoxin
NM
(i) Inhibitor of microtubules(ii) Ability to arrest cell cycle in metaphase
[54]
Viburnum jucundum C.V. Morton
NM
HCT15
<20μg/mL
Acetone
Ursolic acid
Cytotoxic activity
NM
[54]
Hemiangium excelsum (Kunth) A.C.Sm.
NM
HCT15
<10 (μg/mL)
MeOH
PE, EtOAc, MeOH
Cytotoxic activity
NM
[54]
Hyptis pectinata (L.) Poit.
NM
Col2
<4, <20μg/mL
NM
Pectinolide A, pectinolide B, pectinolide C,α-pyrone, boronolide, deacetylepiol-guine
Cytotoxic activity
NM
[54]
H. verticillata Jacq.
NM
Col2
<4,<20μg/mL
NM
Dehydro-β-peltatin, methyl ether dibenzylbutyrolactone, (-)-yatein, 4′-demethyl-deoxypodophyllotoxin
Nonspecific cytotoxic activity
NM
[54]
H. suaveolens (L.)
NM
HF6
2.8-12μg/mL
Chloroform and butanol
β-Apopicropodophyllin
Nonspecific cytotoxic activity
NM
[54]
Salvia leucantha Cav.
Leaf, root, stem
HF6, HT-29, HCT15
14.9, 12.7, 9.9μg/mL
CHCl3
NM
Cytotoxic activity
NM
[54]
Vitex trifolia L.
NM
HCT15
3.5 to <1 (μg/mL)
Hexane and dichloromethane
Salvileucalin B, Hex: leaf, Hex: stem, DCM: leaf, DCM: stem
Cytotoxic activity
NM
[54]
Persea americana Mill.
NM
HT-29
<4μg/mL and <20 μg/mL
Ethanolic
1,2,4-trihydroxynonadecan, 1,2,4-trihydroxyheptadec-16-ene, 1,2,4-trihydroxyheptadec-16-yne
Cytotoxic activity
NM
[54]
Linum scabrellum
Roots, aerial parts
HF6
0.2, 0.5, 2.3μg/mL
Chloroform and butanol
DCM: MeOH, 6MPTOXPTOX
NM
(i) Induction of cell cycle arrest in G2/M(ii) Inhibition of tubulin polymerization
[54]
Phoradendron reichenbachianum (Seem.) Oliv.
NM
HCT15
3.6, 3.9, and 4.3μg/mL
NM
Moronic acid
Cytotoxic activity
NM
[54]
Cuphea aequipetala Cav.
NM
HCT15
18.70μg/mL
Acetone
NM
Cytotoxic inactivity
NM
[54]
Galphimia glauca Cav.
NM
HCT15
0.63, 0.50, 1.99μg/mL
EtOH, MeOH, aqueous
NM
Cytotoxic activity
NM
[54]
Mimulus glabratus Kunth
NM
HF6
12.64μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
Picramnia antidesma Sw.
NM
HCT15
0.6 to 4.5μM
NM
10-Epi-uveoside, uveoside, picramnioside E, picramnioside D
Cytotoxic activity
NM
[54]
Penstemon barbatus (Cav.) Roth
NM
HF6
15.19μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
P. campanulatus (Cav.) Willd.
NM
HF6
6.74μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
Veronica americana Schwein. ex Benth.
NM
HF6
0.169 and 1.46μg/mL
MeOH
NM
Cytotoxic activity
NM
[54]
Zea mays L.
NM
HCT116, SW-480, SW-620
NM
NM
13-Hydroxy-10-oxo-trans-11-octadecenoic acid
Cytotoxic activity
NM
[54]
Colubrina macrocarpa (Cav.) G. Don
NM
HCT15
10, 2.1, 9.1μg/mL
PE, EtOAc, MeOH
NM
Cytotoxic activity
NM
[54]
Coix lacryma-jobi
Seed, endosperm, and hull
HT-29
0.1–1,000μg/mL
Methanolic, hexane
Phytosterols (campesterol, stigmasterol, andβ-sitosterol), gamma-linolenic acid (GLA), arachidonic acid (AA), eicosapentaenoicacid (EPA) and docosahexaenoic acid (DHA), linoleic acid
NM
(i) Influence of signal transduction pathways that involve the membrane phospholipids(ii) Enhancement of ROS generation and decrease of cell antioxidant capacity
[166]
Abutilon indicum
Leaf
HT-29
210μg/mL
Aqueous
Flavonoids (4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl, 2-ethoxy-4-vinylphenol, N,N′-dimethylglycine, lup-20(29)-en-3-one, linolenin, 1-mono-, 9-hexadecanoic acid methyl ester, linolenic acid methyl ester), phenolic (amino acids, terpenoids, fatty acids, methyl palmitoleate)
NM
(i) Increase in the levels of reactive oxygen species and simultaneous reduction in cellular antioxidant, mitochondrial membrane loss, DNA damage, and G1/S phase cell cycle arrest
[167]
Galla rhois
NM
HCT116, HT-29
12.5, 25, 50, 100, 200μg/mL
Aqueous with steaming process
Gallotannins
Increased contents of gallic acid and ellagic acid
(i) Induced apoptosis through the activation of caspases 3, 8, 9(ii) Modulated activation of mitogen and protein kinases, p38, and c-Jun NH2-terminal kinase
[168]
Artemisia annua Linné
Powder
HCT116
20, 30, 40, 60, 80, 100μg/mL
Ethanolic
Phenolic compounds
Inhibited cell viability and increased LDH release
(i) PTEN/p53/PDK1/Akt signal pathways through PTEN/p53 induce apoptosis(ii) Increased apoptotic bodies, caspase 3 and 7 activation(iii) Regulated cytochrome c translocation to the cytoplasm and Bax translocation to the mitochondrial membrane
[169]
Nelumbo nucifera stamen
Powder
HCT116
100, 200, 400μg/mL
Ethanolic crude
NM
NM
(i) Increased the sub-G1 population, mRNA levels of caspases 3 and 8, levels of IκBα and caspase 9(ii) Modulated the Bcl-2 family mRNA expression(iii) Reduced the mRNA levels of NFκB
[170]
Corn silk
NM
LoVo, HCT116
1.25, 2.5, 5, 10, 20μg/mL
Aqueous
Proteins, polysaccharides, flavonoid, vitamins, tannins, alkaloids, mineral salts, steroids
NM
(i) Increase in the Bax, cytochrome c, caspases 3 and 9 levels
[171]
Lycium barbarum L.
Powder
HT-29
1, 2, 3, 4, 5μg/mL
NM
Neoxanthin, all-trans-β-cryptoxanthin, polysaccharides, carotenoids, flavonoids
NM
(i) Upregulation of p53 and p21 expression(ii) Downregulation of the CDK2, CDK1, cyclin A, and cyclin B expression(iii) Arrest in the G2/M phase of cell cycle
[172]
Chrysobalanus icaco L.
Freeze-dried fruit
HT-29
1, 2.5, 5, 10, 20μg/mL
Crude ethyl acetate
Delphinidin, cyanidin, petunidin, and peonidin
NM
(i) Increased intracellular ROS production(ii) Decreased TNF-α, IL-1β, IL-6, and NFκB1 expressions
[173]
Zanthoxylum piperitum De Candolle
Fruit
Caco-2, DLD-1
200μg/mL
Aqueous
NM
NM
(i) Increased the phosphorylation of c-Jun N-terminal kinase (JNK)
[174]
Celtis aetnensis (Tornab.) Strobl (Ulmaceae)
Twigs
Caco-2
5, 50, 100, 250, or 500μg/mL
Methanolic
Flavonoid and triterpenic compounds
NM
(i) Increase in the levels of ROS(ii) Decrease in RSH levels and expression of HO-1
[175]
Rosa canina
Peel and pulp
Caco-2
62.5, 125, 250, 500μg/mL
Total extract (fraction 1), vitamin C (fraction 2), neutral polyphenols (fraction 3), and acidic polyphenols (fraction 4)
Polyphenols
Decreased production of reactive oxygen species (ROS)
NM
[176]
Rhazya stricta
Leaf
HCT116
47, 63, 79, and 95μg/cm2
Crude alkaloid
Alkaloids
NM
(i) Downregulated DNA-binding and transcriptional activities of NFκB and AP-1 proteins(ii) Increase in Bax, caspases 3/7 and 9, p53, p21 and Nrf-2 levels(iii) Decrease in expression of ERK MAPK, Bcl-2, cyclin D1, CDK-4, survivin, and VEGF
[177]
Green coffee
NM
Caco-2
10-1,000μg/mL
NM
5-Caffeoylquinic acid (5-CQA), 3,5-dicaffeoylquinic acid (3,5-DCQA), ferulic acid (FA), caffeic acid (CA), dihydrocaffeic acid (DHCA), dihydroferulic acid (DHFA)
Reduced viability of cancer cells
NM
[178]
Flourensia microphylla
Leaf
HT-29
NM
Ethanolic and acetone
Phenolic compounds
NM
(i) Inhibition of IL-8(ii) Activation of apoptosis by the increment of the Bax/Bcl-2 ratio and expression of TNF family
[179]
∗NM: not mentioned.


## 3.1.2. Studies in Animal Models

The most used animal model is the murine one (Tables2(a) and 2(b)). In particular, studies were carried out above all on HT-29 and HCT116 cells. The effects of the different medicinal plants and their extracts are essentially the same detected in in vitro studies. In particular, plant extracts were able to induce apoptosis and inhibit proliferation and tumor angiogenesis by regulating p53 levels and checkpoint proteins with consequent cell cycle arrest and antiproliferative and antiapoptotic effects on cancerous cells.Table 2
(a)
Efficacy of medicinal plants on colon cancer inin vivo models
Scientific name
Parts used
Model
Dose
Type of extract
Important compounds
Cellular effect
Mechanisms
References
Vitis vinifera
Seed
In vivo (murine)
Caco-2
In vivo: 400–1,000 mg/kgIn vitro: 10–25 μg/mL
Aqueous
Procyanidins
(i) Increased crypt depth and growth-inhibitory effects(ii) Inhibited cell viability(iii) Significantly decreased the histological damage score
Reduced MPO (myeloperoxidase) activity
[180]
Seed
In vivo
HT-29, SW480
5 mg/kg
Aqueous
NM
NM
Decreased VEGF, TNF, MMP-1, MMP-3, MMP-7, MMP-8, MMP-9, and MMP-13 protein expression
[181]
Skin
In vivo
NM
7.5, 30, 60μg/mL
Methanolic
4′-Geranyloxyferulic acid
NM
NM
[30]
Seed
In vivo (murine)
NM
0.12%w/w
NM
Catechin, epicatechin
NM
(i) Suppressed proliferation, sphere formation, nuclear translocation ofβ-catenin and Wnt/β-catenin signaling(ii) Elevated p53, Bax/Bcl-2 ratio, and cleaved PARP and mitochondrial-mediated apoptosis
[31]
Camellia sinensis
Leaf
In vivo (murine)
HT-29
In vitro: 0, 10, 30, 50μMIn vivo: 1.5 mg per day
Aqueous
Catechin, epigallocatechin gallate
1.9-fold increase in tumor endothelial cell apoptosis
Inhibited the ERK-1 and ERK-2 activation, VEGF expression, and VEGF promoter
[182]
In vivo (murine)
HCT116
0.5%
NM
NM
Reduced basement membrane
Inhibition of MMP-9 and VEGF secretion
[183]
In vivo (murine)
Caco-2, HT-29
300μM
Aqueous
Theaflavins (TF-2, TF-3, TF-1)
Induced apoptosis of human colon cancer cells
Inhibition of edema formation correlated to attenuation of COX-2 expression and promoter analysis revealed modulation of NFκB, AP-1, CREB, and/or NF-Il-6 (C/EBP)
[36]
In vivo (murine)
HT115
25μg/mL
Hydroethanolic
Phenolic compounds (p-hydroxyphenyl ethanol, pinoresinol & dihydroxyphenyl ethanol)
NM
Inhibition via reduced expression of a range ofα5 & β1
[184]
Sasa quelpaertensis
Leaf
In vivo
HT-29, HCT116
0, 100, 200, 300 mg/L
Ethanolic
p-Coumaric acid, tricin
Inhibition of colony formation
(i) Nonadherent sphere formation suppressed CD133+ & CD44+ population(ii) Downregulated expression of cancer stem cell markers
[41]
Anoectochilus
NM
In vivo
CT26
Oral dose of 50 & 10 mg/mouse per day
Aqueous
Kinsenoside
Stimulated proliferation of lymphoid tissues
Activation of phagocytosis of peritoneal macrophages
[185]
Purple-fleshed potatoes
Fruit
In vivo
Colon cancer stem cells
5.0μg/mL
Ethanol, methanol, ethyl acetate
Anthocyanin,β-catenin, cytochrome c
Reduction in colon CSCs number and tumor incidence
(i) Increase in cytochrome c levels from p53 status and maybe mitochondria-mediated apoptosis(ii) Suppressed levels of cytoplasmic and nuclear β-catenin
[58]
Phaseolus vulgaris
Leaf
In vivo
HT-29
Nm
Ethanolic
Polysaccharides, oligosaccharides
Induction of apoptosis and inhibit proliferation
(i) Inactivation of the retinoblastoma phosphoprotein(ii) Induced G1 arrest(iii) Suppression of NF-jb1(iv) Increase in EGR1 expression
[59]
Rosmarinus officinalis L.
Leaf
In vivo
HT-29
SC-RE 30μg/mL and CA 12.5 μg/mL
Ethanolic
Polyphenols (carnosic acid (CA) and carnosol)
(i) Activation of Nrf2 transcription factor(ii) Activated common regulators, such as XBP1 (Xbp1) gene, SREBF1/SREBF2 (Srebp1/2), CEBPA and NR1I2 (Pxr) genes
Leaf
In vivo (rat)
NM
NM
Ethanolic
Rosmanol and its isomers, carnosol, rosmadial, carnosic acid, and 12-methoxycarnosic acid, carnosic acid, carnosol
Interactions with the gut microbiota and by a direct effect on colonocytes with respect to the onset of cancer or its progression
NM
Wasabia japonica
Rhizomes
In vivo
COLO 205
5 mg/mL
Methanolic
6-(Methylsulfinyl)hexyl isothiocyanate
Anticolon cancer properties through the induction of apoptosis and autophagy
(i) Activation of TNF-α, Fas-L, caspases(ii) Truncated Bid and cytochrome c(iii) Decreased phosphorylation of Akt and Mtor(iv) Promoted expression of microtubule-associated protein 1 light chain 3-II and AVO formation
[186]
Zingiberaceae
Rhizome
HT-29
HT-29
5 g/kg
Dichloromethanic
Turmerone
Suppressed the proliferation of HT-29 colon cancer cells
(i) LDH release(ii) ROS generation(iii) Collapse in mitochondrial membrane potential(iv) Cytochrome c leakage(v) Activation of caspase 9 and caspase 3
[187]
Panax quinquefolius
Root
In vivo (murine)
NM
30 mg/kg
Ethanolic
Ginsenosides (protopanaxadiol or protopanaxatriol)
Attenuated azoxymethane/DSS-induced colon carcinogenesis by reducing the colon tumor number and tumor load
(i) Reduced experimental colitis(ii) Attenuated on AOM/DSS-induced colon carcinogenesis(iii) Proinflammatory cytokines activation(iv) Suppressed DSS(v) Downregulated inflammatory cytokine gene expression
[188]
Myrtaceae
Leaf
In vivo (murine)
HCT116
100μg/mL (in vitro) 200 and 100 μg/disc (in vivo)
Methanolic
Phenolics, flavonoids, betulinic acid
Inhibition of tumor angiogenesis
(i) Inhibition of angiogenesis of tube formation on Matrigel matrix and HUVECS migration (in vitro)(ii) Decreased nutrient and oxygen supply and consequently tumor growth and tumor size (in vivo)(iii) Increased extent of tumor necrosis
[82]
Spica prunellae
Leaf
In vivo
HT-29
200 mg/mL (in vitro), 600 mg/mL (in vivo)
Ethanolic
Rosmarinic acid
Induction of apoptosis and inhibition of cell proliferation and tumor angiogenesis
(i) Induced apoptosis(ii) Inhibited cancer cell proliferation and angiogenesis STAT3 phosphorylation(iii) Regulated expression of Bcl-2, Bax, cyclin D1, CDK4, VEGF-A, and VEGFR-2 (in vivo)
[83]
Gymnaster koraiensis
Aerial part
In vivo (murine)
NM
500μmol/kg
Ethanolic
Gymnasterkoreaynes B, C, E, 2,9,16-heptadecatrien-4,6-dyne-8-ol
Anti-inflammatory and cancer preventive activities
(i) Significant decrease in expression of COX-2(ii) Increase in serum IL-6
[189]
Allium fistulosum
Edible portions
In vivo (murine)
CT26
50 mg/kg b.w.
Hot water
p-Coumaric acid, ferulic acid, sinapic acid, quercitrin, isoquercitrin, quercetol, kaempferol
Suppression of tumor growth and enhanced survival rate of test mice
(i) Decreased expression of inflammatory molecular markers(ii) Downregulated expression of MMP-9 and ICAM(iii) Metabolite profiling and candidate active phytochemical components
[190]
Annona squamosa Linn
Leaf
In vivo (animal)
HCT116
8.98μg/mL
Crude ethyl acetate
Acetogenins (annoreticuin & isoannoreticuin) and alkaloids dopamine, salsolinol, and coclaurine
(i) Inhibited growth and proliferation of tumor cells
Reactive oxygen species (ROS) formation, lactate dehydrogenase (LDH) release, and caspases 3/7, 8, 9 activation
[191]
Eupatorium cannabinum
Aerial parts
In vivo (murine)
HT-29
25μg/mL
Ethanolic
Pyrrolizidine alkaloids (senecionine, senkirkine, monocrotaline, echimidine)
Cytotoxicity against colon cancer cells
(i) Upregulation of p21 and downregulation of NCL, FOS, and AURKA, indicating reduced proliferation capacity(ii) Mitotic disruption and nonapoptotic cell death via upregulation of Bcl-xL
[96]
Flacourtia indica
Aerial parts
In vivo (murine)
HCT116
500μg/mL
Methanolic
Phenolic glucoside (flacourticin, 4′-benzoylpoliothrysoside)
Antiproliferative and proapoptotic effects in HCT116 cells
Apoptosis via generation of ROS and activation of caspases (PARP)
[192]
Sorghum bicolor
The dermal layer of stalk
In vivo (murine)
HCT116 & colon cancer stem cells
>16 and 103μg/mL
Phenolic, acetone
Apigeninidin & luteolinidin
Antiproliferative effect
(i) Target p53-dependent and p53-independent pathways
[97]
Gleditsia sinensis
Thorn
In vivo (murine)
HCT116
800μg/mL
Aqueous
Flavonoid, lupine acid, ellagic acid glycosides
Inhibited proliferation of colon cancer
(i) Increased p53 levels(ii) Downregulation of the checkpoint proteins, cyclin B1, Cdc2, and Cdc25c
[90]
Thorn
In vivo (murine)
HCT116
600μg/mL
Ethanolic
NM
Inhibitory effect on the proliferation of human colon cancer HCT116 cells
(i) Caused G2/M phase cell cycle arrest
[91]
Zingiber officinale
Rhizome
In vitro/in vivo (murine)
HCT116
5μM
Ethanolic
6-Paradol, 6- and 10-dehydrogingerdione, 6- and 10-gingerdione, 4-, 6-, 8-, and 10-gingerdiol, 6-methylgingerdiol, zingerone, 6-hydroxyshogaol, 6-, 8-, 10-dehydroshogaol, diarylheptanoids
Inhibitory effects on the proliferation of human colon cancer cells
(i) Arrest of G0/G1 phase(ii) Reduced DNA synthesis(iii) Induced apoptosis
[103]
Cucumaria frondosa
The enzymatically hydrolyzed epithelium of the edible
In vivo (murine)
HCT116
<150μg/mL
Hydroalcoholic
Monosulphated triterpenoid glycoside frondoside A, the disulphated glycoside frondoside B, the trisulphated glycoside frondoside C
(i) Inhibition at S and G2-M phase with a decrease in Cdc25c(ii) Increase in p21WAF1/CIP
(i) Inhibition the growth of human colon(ii) Apoptosis associated with H2AX phosphorylation and caspase 2
[105]
Rolandra fruticosa
Leaf & twigs
In vivo (murine)
HT-29
10 and 5 mg/kg/day
Methanolic
Sesquiterpene lactone (13-acetoxyrolandrolide)
Antiproliferative effect against human colon cancer cells
(i) Inhibition of the NFκB pathway, subunit p65 (RelA) and upstream mediators IKKβ and oncogenic K-ras
[106]
Cydonia oblonga Miller
Leaf & fruit
In vivo (murine)
Caco-2
250–500μg/mL
Methanolic
Phenolic compound (flavonol and flavone heterosides, 5-O-caffeoylquinic acid)
Antiproliferative effect against human kidney and colon cancer cells
(i) Suppression of NFκB activation, activator (AP-1), mitogen-activated protein kinases, namely, PKC, (GFR)-mediated pathways(ii) Cell cycle arrest(iii) Induction of apoptosis, antioxidant, and anti-inflammatory effects
[107]
Sedum kamtschaticum
Aerial part
In vivo (murine)
HT-29
0–0.5 mg/mL
Methanolic
Buddlejasaponin IV
Induced apoptosis in HT-29 human colon cancer cells
(i) Induced apoptosis via mitochondrial-dependent pathway triggered by downregulation of Bcl-2 protein levels, caspase 3 activation, and subsequent PARP cleavage
[109]
Ganoderma lucidum
Caps & stalks
In vivo (murine)
HT-29
0-0.1 mg/mL
Triterpene extract (hot water) extract
Polysaccharides (mainly glucans & glycoproteins), triterpenes (ganoderic acids, ganoderic alcohols, and their derivatives)
Cytokine expression inhibited during early inflammation in colorectal carcinoma
Induced autophagy through inhibition of p38 mitogen-activated kinase and activation of farnesyl protein transferase (FPT)
[193]
Ginkgo biloba
Fruit & leaf
In vivo (murine)
HT-29
20–320 mg/L
Aqueous
Terpene lactones and flavonoid glycosides
Inhibited progression of human colon cancer cells induced HT-29 cell apoptosis
(i) Activation in caspase 3, reduction in Bcl-2 expression, and elevation in p53 expression
[112]
Rubus occidentalis
Fruit
In vivo (murine)
JB6 Cl 41
25μg/mL
Methanolic
β-Carotene, α-carotene, ellagic acid, ferulic acid, coumaric acid
Inhibited tumor development
(i) Impaired signal transduction pathways leading to activation of AP-1 and NFB RU-ME fraction
[194]
Oryza sativa
Seed
In vivo (murine)
HT-29, SW 480, HCEC
100μg/mL
Ethyl acetate extract
Phenolic compound (tricin, ferulic acid, caffeic acid, and methoxycinnamic acid)
Inhibited growth of human colon cancer cells
(i) Induction of apoptosis by enhanced activation of caspases 8 and 3(ii) Decreased the number of viable SW480 and HCEC cells
[113]
Cistanche deserticola
Dried stem
In vivo (murine)
SW480
In vivo: 0.4 g/kg/dayIn vitro: 100 mg/mL
Aqueous
Polysaccharides, phenylethanoid glycosides
Decreased mucosal hyperplasia and helicobacter infection
(i) Increased number of splenic macrophages and NK cells(ii) Decreased frequency of hyperplasia and H. hepaticus infection of the intestine
[133]
Rehmannia glutinosa
NM
In vivo (male C57BL6 mice and Sprague-Dawley rats)
CT26
28 mg/kg
NM
Catalpol
(i) Inhibited proliferation, growth, and expression of angiogenic markers
(i) VEGF, VEGFR2, HIF-1α, bFGF inhibited the expressions of inflammatory factors such as IL-1β, IL-6, and IL-8
[143]
Olea europaea
Olive mill wastewater
In vivo (murine)
NM
NM
Methanolic
Hydroxytyrosol
Interferes with tumor cell growth
NM
[195]
Leaf
In vivo (xenograft model) (murine)
HCT116, HCT8
0, 5, 10, 20, 30, 50, and 70μg/mL
Phenolic
Oleuropein and hydroxytyrosol
NM
(i) Activation of caspases 3, 7, and 9(ii) Decrease of mitochondrial membrane potential and cytochrome c release(iii) Increase in intracellular Ca2+ concentration
[196]
Ginkgo biloba L.
Leaf
In vivo (rat)
NM
0.675 and 1.35 g/kg
Methanolic
Flavonoid glycosides, terpene lactones, and ginkgolic acids
(i) Suppressed tumor cell proliferation, promoted apoptosis, and mitigated inflammation
NM
[197]
Rhus trilobata Nutt.
NM
In vivo (hamster)
NM
400 mg/kg, 100 mg/kg
Aqueous
Tannic acid, gallic acid
Cytotoxic activity
NM
[54]
Annona diversifolia Saff.
NM
In vivo (mice)
SW-480
1.5, 7.5 mg/kg/day
NM
Laherradurin
Cytotoxic activity
NM
[54]
A. muricata L.
NM
In vivo (rat)
NM
250/500 mg/kg
EtOAc
A, B, and C, and cis- and trans-annomuricin-D-ones
Cytotoxic activity
NM
[54]
Plumeria acutifolia Poir.
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Lasianthaea podocephala (A. Gray) K. M. Becker
NM
In vivo (hamster)
NM
200 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Flourensia cernua DC.
NM
In vivo (hamster)
NM
350 mg/kg/day
Aqueous
Flavonoids, sesquiterpenoids, monoterpenoids, acetylenes, p-acetophenones, benzopyrans, benzofurans
Cytotoxic activity
NM
[54]
Ambrosia ambrosioides (Cav.) W. W. Payne
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Alnus jorullensis Kunth
NM
In vivo (hamster)
NM
175 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Dimorphocarpa wislizeni (Engelm.) Rollins
NM
In vivo (hamster)
NM
100 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Euphorbia pulcherrima Willd. ex Klotzsch
NM
In vivo (hamster)
NM
200 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Acalypha monostachya Cav.
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Crotalaria longirostrata Hook. & Arn.
NM
In vivo (hamster)
NM
400 mg/kg/day, 350 mg/kg/day
EtOH-CHCl3
NM
Cytotoxic activity
NM
[54]
Asterohyptis stellulata (Benth.) Epling
NM
In vivo (hamster)
NM
50 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Acacia constricta A. Gray
NM
In vivo (hamster)
NM
400 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Holodiscus dumosus A. Heller
NM
In vivo (hamster)
NM
350 mg/kg/day
Aqueous
NM
Cytotoxic activity
NM
[54]
Butea monosperma
Flower
In vivo (rat)
HT-29
150 mg/kg
n-Butanol extract
Isocoreopsin, butrin, and isobutrin
Free radical scavenging and anticancer activities
NM
[198]
Taraxacum spp.
Root
In vivo (xenograft murine model)
HT-29, HCT116
40 mg/kg/day
Aqueous
α-Amyrin, β-amyrin, lupeol, and taraxasterol
Induced programmed cell death
NM
[199]
∗NM: not mentioned.
(b)
Other effects of medicinal plants inin vivo models
Scientific name
Parts used
Model
Dose
Type of extract
Important compounds
Cellular effect
Mechanisms
References
Allium sativum
Root
In vivo (murine)
NM
2.4 mL of daily
Ethanolic
Allicin, S-allylmercaptocysteine
Significantly suppressed both the size and number of colon adenomas
Enhancement of detoxifying enzymes: SAC and GST activity
[200]
Olea europaea
Fruit
In vivo
Caco-2
50μM
Aqueous
Phenolic compounds, authentic hydroxyl tyrosol (HT)
(i) Effect of OPE and HT on CB1 associated with reduced proliferation of Caco-2 cells(ii) Increase in CB1 expression in the colon of rats receiving dietary EVOO
Increase in Cnr1 gene expression, CB1 protein levels
[201]
In vivo (murine)
HT115
25μg/mL
Hydroethanolic
Phenolic compounds (p-hydroxyphenyl ethanol, pinoresinol & dihydroxyphenyl ethanol)
NM
Inhibition via reduced expression of a range ofα5 & β1
[184]
Origanum vulgare L.
Leaf
In vivo (murine)
NM
20, 40, 60 mg·kg−1
Aqueous
Rosmarinic acid, caffeic acid, flavonoids
Antioxidant status
(i) Increased LPO products and activity of SOD and CAT enzymes and GST and GPx activity(ii) Antioxidant and anticarcinogenic effect
[202]
Hazelnut
Skin
In vivo
NM
The flow rate 0.21 mL/min and injection volume 9.4μL
Aqueous
Flavan-3-ols, in monomeric and polymeric forms, and phenolic acids
(i) Decreased circulating levels of free fatty acids and triglycerides(ii) Higher excretion of bile acid
Increase of the total antioxidant capacity of plasma
[203]
Apples and apple juice
Fruit
In vivo
NM
90 mg/L
Aqueous
Phenolic acids, flavonoids, tannins, stilbenes, curcuminoids
NM
NM
[204]
Grifola frondosa
Fruit
In vivo (murine)
HT-29
10 ng/mL
Aqueous
Phenolic compounds (pyrogallol, caffeic acid, myricetin, protocatechuic acid, etc.)
Inhibition of TNBS-induced rat colitis
(i) Induced cell cycle progression in G0/G1 phase and apoptotic death
[104]
Ruta chalepensis
Leaf
In vivo (human)
NM
250μg/mL
Ethanolic
Rutin, gallic acid, catechin hydrate, naringin
Oxidative profile in patients with colon cancer
NM
[205]
Cannabis sativa
Dry flower & leaf
In vivo (murine)
DLD-1 and HCT116
0.3–5μM
Methanolic
Cannabidiol, phytocannabinoids
NM
(i) Reduced cell proliferation in a CB1-sensitive and AOM-induced preneoplastic lesions and polyps(ii) Inhibition of colorectal cancer cell proliferation via CB1and CB2 receptor activation
[121]
Melia toosendan
Fruit
In vivo (murine)
SW480, CT26
0, 10, 20, 30, 40, 50μg/mL
Ethanolic
Triterpenoids, flavonoids, polysaccharide, limonoids
NM
(i) Inhibited cell proliferation of SW480 and CT26 by promoting apoptosis as indicated by nuclear chromatin condensation and DNA fragmentation(ii) Induced caspase 9 activity which further activated caspase 3 and poly(ADP-ribose) polymerase cleavage, leading the tumor cells to apoptosis
[123]
Smallanthus sonchifolius
Root
In vivo (murine)
NM
73.90, 150.74, 147.65, and 123.26 mg/kg
Aqueous
Fructans
NM
Reduction incidence of colon tumors expressing alteredβ-catenin
[206]
Punica granatum
Peel
In vivo (adult male Wistar rats)
NM
4.5 g/kg
Methanolic
Gallic acid, protocatechuic acid, cateachin, rutin, ellagic acid, punicalagin
NM
(i) Reduction in TGF-β, Bcl-2, EGF, CEA, CCSA-4, MMP-7 and in COX-2, cyclin D1, survivin content(ii) Downregulated expression of β-catenin, K-ras, c-Myc genes
[207]
Linum usitatissimum
Seed
In vivo (male Sprague-Dawley rats)
NM
500 mg/kg
Alkaline
Secoisolariciresinol diglucoside, carbohydrates, proteins, and tannins
Reduced the serum fasting glucose levels
Significantly reduced the HbA1c, insulin levels, and proinflammatory cytokines
[208]
Diospyros kaki
Fruit
In vivo (male CD-1 mice)
NM
15 mg/kg
Hydroacetone
Polyphenol
(i) Decreased attenuation of colon length in diarrhea severity(ii) Reduced mortality rate(iii) Reduction of the extent of visible injury (ulcer formation) and of mucosal hemorrhage
Decreased expression of COX-2 and iNOS in the colonic tissue
[147]
Muntingia calabura
Leaf
In vivo (rat)
NM
50, 250, 500 mg/kg
Methanolic
Rutin, gallic acid, ferulic acid, and pinocembrin
Reduction of the colonic oxidative stress, increasing the antioxidants levels possibly via the synergistic action of several flavonoids
NM
[209]
Portulaca oleracea
NM
In vivo (murine)
HT-29 CSCs
2.25μg/mL
Alcoholic
NM
Regulatory and target genes that mediate the Notch signal transduction pathway
Inhibition of expression of the Notch1 andβ-catenin genes
[161]
Aloe vera
Gel
In vivo (murine)
NM
400 mg/kg/day
Gel
Polysaccharides
NM
(i) Via inhibition of the cell cycle progression(ii) Induction of cellular factors, such as extracellular signal-regulated kinases 1/2, cyclin-dependent kinase 4, and cyclin D1; on the other hand, PAG increased the expression of caudal-related homeobox transcription factor 2
[210]
Artemisia annua Linné
Powder
In vivo (xenograft murine model)
HCT116
20, 40 mg/kg/day
Ethanolic
Phenolic compounds
NM
(i) Induced apoptosis via PTEN/p53/PDK1/Akt signal pathways through PTEN/p53(ii) Inhibited cell viability and increased LDH release and apoptotic bodies, caspase 3 and 7 activation, and reduced mitochondria membrane potential(iii) Regulated cytochrome c translocation to the cytoplasm and Bax translocation to the mitochondrial membrane(iv) Regulation of proteins
[169]
Hordeum vulgare
Powder
In vivo (xenograft murine model)
HT-29
2 g/kg and 1 g/kg
Aqueous (fermented)
β-Glucan, protein, amino acids, phenolic compounds
NM
(i) Promoted tumor apoptosis by upregulating the mRNA expression of Bax and caspase 3 and downregulating the mRNA expression of Bcl-2 and cyclin D1(ii) Decreased mRNA expression of Bcl-2 and cyclin D1(iii) Upregulated expressions levels of Bax and caspase 3
[211]
Dendrophthoe pentandra
Leaf
In vivo (murine)
NM
125, 250, 500 mg/kg
Ethanolic
Quercetin-3-rhamnose
NM
(i) Decreased the levels of IL-22, MPO levels, proliferation of epithelial cells(ii) Inhibited S phase of the cell cycle(iii) Upregulated p53 wild-type gene expression
[212]
Aquilaria crassna
Stem, bark
In vivo (murine)
HCT116
2,000 mg/kg/day100, 200 mg/kg
NM
Resin and essential oils
NM
NM
[213]
Berberis integerrima
NM
In vivo (murine)
NM
50 and 100 mg/kg
Hydroalcoholic
NM
NM
NM
[214]
Salix aegyptiaca
Bark
In vivo (murine)
NM
100 and 400 mg/kg
Ethanolic
Catechin, catechol, and salicin
NM
Decreased level of EGFR, nuclearβ-catenin, and COX-2
[215]The main mechanisms of action of medicinal plants are summarized in Figure1.Figure 1
Cell damage and cancer trigger p53 activation. The p53 protein activates the apoptotic protein Bax. Bax inhibits the antiapoptotic protein Bcl-2. During apoptosis, cytochrome c is released from mitochondria. To activate the Apaf-1 protein, the interaction between these proteins and cytochrome C is necessary. Pro-caspase 9 attaches to Apaf-1 and activates caspase 9. Caspase 9 activates caspases 3 and 7 and apoptosis occurs.Inin vitro studies, it has been found that grapes, which contain substantial amounts of flavonoids and procyanidins, play a role in reducing the proliferation of cancer cells by increasing dihydroceramides and p53 and p21 (cell cycle gate keeper) protein levels. Additionally, grape extracts triggered antioxidant response by activating the transcriptional factor nuclear factor erythroid 2-related factor 2 (Nrf2) [27].Grape seeds contain polyphenolic and procyanidin compounds, and their reducing effects on the activity of myeloperoxidase have been shown inin vitro and in vivo studies. It has been suggested that grape seeds could inhibit the growth of colon cancer cells by altering the cell cycle, which would lead eventually to exert the caspase-dependent apoptosis [180].Another plant that attracted researchers’ attention was soybean, which contain saponins. After 72 h of exposure of colon cancer cells to the soy extract, it was found that this extract inhibited the activity and expression of protein kinase C and cyclooxygenase-2 (COX-2) [34]. The density of the cancer cells being exposed to the soy extract significantly decreased. Soybeans can also reduce the number of cancer cells and increase their mortality, which may be due to increased levels of Rab6 protein [216].Green tea leaves have also attracted the researchers’ attention in these studies. Green tea leaves, with high levels of catechins, increased apoptosis in colon cancer cells and reduced the expression of the vascular endothelial growth factor (VEGF) and its promoter activity inin vitro and in vivo studies. The extract increased apoptosis (programmed cell death) by 1.9 times in tumor cells and 3 times in endothelial cells compared to the control group [182]. In another in vitro study, the results showed that green tea leaves can be effective in the inhibition of matrix metalloproteinase 9 (MMP-9) and in inhibiting the secretion of VEGF [183].Garlic was another effective plant in this study. Its roots have allicin and organosulfur compounds. In anin vitro study, they inhibited cancer cell growth and induced apoptosis through the inhibition of the phosphoinositide 3-kinase/Akt pathway. They can also increase the expression of phosphatase and tensin homolog (PTEN) and reduce the expression of Akt and p-Akt [32]. Garlic roots contain S-allylcysteine and S-allylmercaptocysteine, which are known to exhibit anticancer properties. The results of a clinical trial on 51 patients, whose illness was diagnosed as colon cancer through colonoscopy, and who ranged in age from 40 to 79 years, suggest that the garlic extract has an inhibitory effect on the size and number of cancer cells. Possible mechanisms suggested for the anticancer effects of the garlic extract are both the increase of detoxifying enzyme soluble adenylyl cyclase (SAC) and an increased activity of glutathione S-transferase (GST). The results suggest that the garlic extract stimulates mouse spleen cells, causes the secretion of cytokines, such as interleukin-2 (IL2), tumor necrosis factor-α (TNF-α), and interferon-γ, and increases the activity of natural killer (NK) cells and phagocytic peritoneal macrophages [200].The results ofin vitro studies on olive fruit showed that it can increase peroxide anions in the mitochondria of HT-29 cancer cells due to the presence of 73.25% of maslinic acid and 25.75% of oleanolic acid. It also increases caspase 3-like activity up to 6 times and induces programmed cell death through the internal pathway [217]. Furthermore, the olive extract induces the production of reactive oxygen species (ROS) and causes a quick release of cytochrome c from mitochondria to cytosol.The pomegranate fruit contains numerous phytochemicals, such as punicalagins, ellagitannins, ellagic acid, and other flavonoids, including quercetin, kaempferol, and luteolin glycosides. The results of anin vitro study indicate the anticancer activity of this extract through reduction of phosphorylation of the p65 subunit and subsequent inhibition of nuclear factor-κB (NFκB). It also inhibits the activity of TNF receptor induced by Akt, which is needed for the activity of NFκB. The fruit juice can considerably inhibit the expression of TNF-α-inducing proteins (Tipα) in the COX-2 pathway in cancer cells [43]. The effective and important compounds in pomegranate identified in these 104 studies are flavonoids, polyphenol compounds, such as caffeic acid, catechins, saponins, polysaccharides, triterpenoids, alkaloids, glycosides, and phenols, such as quercetin and luteolin, and kaempferol and luteolin glycosides.In a systematic review of the plants being studied, some mechanisms were mainly common, including the induction of apoptosis by means of an increase of expression and levels of caspase 2, caspase 3, caspase 7, caspase 8, and caspase 9 in cancer cells, increasing the expression of the proapoptotic protein Bax and decreasing the expression of the antiapoptotic proteins.Many herbal extracts block specific phase of the cell cycle. For instance, the extract prepared from the leaves ofAnnona muricata inhibits the proliferation of colon cancer cells and induces apoptosis by arresting cells in the G1 phase [53]. They can also prevent the progress of the G1/S phase in cancer cells [74]. In general, the herbal extracts reported here have been able to stop cancer cells at various stages, such as G2/M, G1/S, S phase, G0/G1, and G1 phase, and could prevent their proliferation and growth.Other important anticancer mechanisms are the increase of both p53 protein levels and transcription of its gene. Even the increase of p21 expression is not without effect [137]. In an in vitro study on the Garcinia mangostana roots, the results were indicative of the inhibitory effect of the extract of this plant on p50 and P65 activation [93]. Moreover, reduction of cyclin D1 levels and increase of p21 levels are among these mechanisms [137], as well as inhibition of NFκB and reduction of the transcription of its genes, which contribute to reduce the number of cancerous cells [127]. Other important anticancer mechanisms are the inhibition of COX-2, as well as the reduction of the protein levels in this pathway [34]. In addition to this, in some cases, the inhibition of MMP-9 can be mentioned as the significant mechanism of some herbal extracts to kill cancer cells [183].


## 4. Conclusion and Perspectives

The findings of this review indicate that medicinal plants containing various phytochemicals, such as flavonoids, polyphenol compounds, such as caffeic acid, catechins, saponins, polysaccharides, triterpenoids, alkaloids, glycosides, and phenols, such as quercetin and luteolin, and kaempferol and luteolin glycosides, can inhibit tumor cell proliferation and also intduce apoptosis.Plants and their main compounds affect transcription and cell cycle via different mechanisms. Among these pathways, we can point to induction of superoxide dismutase to eliminate free radicals, reduction of DNA oxidation, induction of apoptosis by inducing a cell cycle arrest in S phase, reduction of PI3K, P-Akt protein, and MMP expression, reduction of antiapoptotic Bcl-2, Bcl-xL proteins, and decrease of proliferating cell nuclear antigen (PCNA), cyclin A, cyclin D1, cyclin B1, and cyclin E. Plant compounds also increase the expression of both cell cycle inhibitors, such as p53, p21, and p27, and BAD, Bax, caspase 3, caspase 7, caspase 8, and caspase 9 proteins levels. In general, this study showed that medicinal plants are potentially able to inhibit growth and proliferation of colon cancer cells. But the clinical usage of these results requires more studies on these compounds inin vivo models. Despite many studies’ in vivo models, rarely clinical trials were observed among the studies. In fact, purification of herbal compounds and demonstration of their efficacy in appropriate in vivo models, as well as clinical studies, may lead to alternative and effective ways of controlling and treating colon cancer.



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*Source: 2075614-2019-12-04.xml*