Genistein-triggered anticancer activity against liver cancer cell line HepG2 involves ROS generation, mitochondrial apoptosis, G2/M cell cycle arrest and inhibition of cell migrationand inhibition of cell migration
Authors: Qian Zhang
Juan Bao
Jiehua Yang
Journal: Archives of Medical Science (2019)
Discipline: CLINICAL MEDICINE
Keywords: HepG2; anticancer; genistein; liver cancer.
Affiliations: Second Department of Hepatobiliary Surgery, Qujing Second People's Hospital, Qujing, China., Department of Cerebrovascular Disease, The Second Affiliated Hospital, Kunming Medical University, Kunming, China.

Abstract:  Introduction: Liver cancer is one of the most common malignancies across the globe and one of the major causes of cancer-related mortality. With limited available treatment options, there is an urgent need to look for new available options. Genistein is an important plant flavonoid and has been shown to possess tremendous pharmacological potential. The objective of the present study was therefore to evaluate the anticancer effect of the genistein. Material and methods: The antiproliferative activity and IC50 of genistein were determined by MTT assay. Reactive oxygen species (ROS) and cycle distribution were investigated by flow cytometry. Apoptosis was detected by DAPI and annexin V/IP staining. Cell migration was investigated by wound healing assay. Protein expression was estimated by western blotting. Results: MTT assay revealed that genistein reduced the cell viability of HepG2 cancer cells in a dose-dependent manner. Genistein also reduced the colony forming potential of the HepG2 cell concentration dependently. The IC50 of genistein was found to be 25 μM. Genistein caused G2/M cell cycle arrest and G2/M cells increased from 4.2% in the control to 56.4% at 100 μM concentration. Genistein prompted generation of significant (p &lt; 0.01) amounts of ROS, ultimately favouring cell death. Genistein also triggered apoptosis which was associated with upregulation of cytosolic cytochrome c, Bax, cleaved caspase 3 and 9 expression and downregulation of Bcl-2 expression in HepG2 cells. Conclusions: We propose that genistein exhibits significant anticancer activity against liver cancer and therefore may prove beneficial in the management of liver cancer. 

Retracted: No

PubPeer comments:
 Fig 2A.  Colony formation assay of HepG2 cells: A u2013 inhibition of colony formation . ![] 25 and 50 microM panels are the same. For comparison: Fig 1C from [Chen et al (2019)] . ![] Fig 3 from [Zhang, Li Ji (2018)] , and Figure 1C from [Huang, Xie Liu (2019)] . ![] , 
 #1 After I posted the marked figure, just saw, although you didn t mark it, you did describe in your text. ![file] , 
 *[left]* Figure 5 from [Zhang, Bao Yang (2019)] .  Detection of apoptosis by DAPI staining at indicated doses of genistein . *[right]* Fig 3 from [Duan et al (2019)] .  Figure 3. Mulberroside-A induces dose-dependent apoptotic cell death as indicated by DAPI staining. The upper two-thirds of each panel from the left is also the lower half of a panel from the right. ![file] , 
 Figure 7.  The effect of indicated doses of genistein on the expression of caspase-dependent mitochondrial apoptosis pathway proteins in HepG2 cells. Representative images of cytochrome c, Bax, Bcl-2, PARP, cleaved caspase 9 and cleaved caspase 3 protein expression detected by western blot. u03b2-actin was used as a control. ![] For comparison: *[left]* Figure 5 from [Li, Yang Shen (2019)] .  The effect of indicated doses of gentiopicroside on Bax and Bcl-2 expression as indicated by western blotting. *[middle]* Figure 5 from [Deng Li (2019)] .  Effect of linderalactone on the cell cycle phase distribution of A549 cells as depicted by Western blotting. *[right]* Fig. 7 from [Bi et al (2018)] .  Effect of indicated doses of genistein ... (B) protein expression of survivin, cyclin D1 and ALDH1A1 . ![] , 
 #3 Fig 5 again *[left]]* ![] Another sighting! Fig 4 from [Zhang et al (2018)] : ![] , 
 #4 Fig 7 again *left]*. The PARP band appears to come from Fig 5D of [Bi et al (2018)] *[right]*. ![file] -------------------------- No, wait - an earlier appearance. Fig 2C of  Hispidulin Induces Apoptosis Through Mitochondrial Dysfunction and Inhibition of P13k Akt Signalling Pathway in HepG2 Cancer Cells ([Gao et al 2014] ). ![file] ,