# Presentation: 592717

## Tea and Tea Polyphenols in Cancer Prevention

**Tea and Tea Polyphenols in Cancer Prevention**

- Sharon Ross, PhD, MPH
- _[rosssha@mail.nih.gov](mailto:rosssha@mail.nih.gov)_
- Nutritional Science Research Group,
- Division of Cancer Prevention

## Tea (Camellia sinensis)

**Green Tea**

**Black Tea**

- 30-40%   Catechins
-     3-6%	  Caffeine
- ~310 mg polyphenols
- per 6 ounces

- 3-10%	 Catechins
-   2-6%	 Theaflavins
- > 20%	 Thearubigens
-   3-6%    Caffeine
- ~340 mg polyphenols
- per 6 ounces

- Dried
- Tea Leaves

- Crushed tea leaves
- Polyphenol oxidase
- Oxidation,
- Polymerization

** **

** ****Yang, CS. Personal Communication**

## Increased Concentration of Catechins Following Black Tea Consumption

- Plasma Levels
- (nmol/L)

- Urinary Excretion
- (nmol/h)

- Fecal Excretion
- (****mol)

**Epigallocatechin**
**Epicatechin**
**Epigallocatechin Gallate**
**Epicatechin Gallate**

**Warden BA, et al. J Nutr 2001;131:1731-1737**

## Epidemiological Studies of

**Epidemiological Studies of**
**Tea and Cancer**

- Ecologic, case-control and cohort studies have been performed.
- Many performed as secondary analyses.
- Little information on precision of tea intake.
- Several cancer sites investigated: bladder and urinary tract, breast, colon and rectum, esophagus, kidney, liver, lung, nasopharynx, pancreas, stomach and uterus (with mixed results).

## Recent Epidemiological Studies of Tea

**Recent Epidemiological Studies of Tea**
**And Gastric/Stomach Cancer**

- Country (Tea type)

- Study Type

- Risk/Association

- Reference

- China
- (green tea)
- India
- (unknown)
- Japan
- (green tea)
- Japan
- (green tea)
- Japan
- (green tea)
- China
- (unknown**)
- Japan
- (green tea)
- Japan
- (green tea)

- Case-
- control
- Case-
- control
- Prospective
- cohort
- Prospective
- cohort
- Prospective
- cohort
- Nested case-
- control
- Prospective
- cohort
- Nested case-
- control

- Decrease
- Decrease
- No association
- No association*
- No association
- Decrease
- Decrease***
- No association

- Setiawan
- 2001
- Rao 2002
- Tsubono
- 2001
- Hoshiyama 2002
- Fujino 2002
- Sun 2002
- Sasazuki 2004
- Hoshiyama 2004

*** Deaths   **Urinary polyphenols/metabolites ***distal gastric, women only**

## Effects of Tea on Human Oral Precancerous Lesions

**A double-blind intervention trial of 59 patients with oral mucosa leukoplakia**
**Twenty-nine patients received tea administered orally and topically; 30 patients received placebo treatment**
**After 6 months, the size of oral lesion decreased in 38% of the treated group and in 10% of the placebo group;  the lesion increased in 3.4% of the treated group and in 6.7% of the placebo group**
**The incidence of micronucleated exfoliated oral mucosa cells in the treated group (0.54%) was lower than the control group (1.13%)**

***Li et al.  Proc. Soc. Expr. Biol. Med. 220: 218-224, 1999.***

## Effect of Increased Tea Consumption on Oxidative DNA Damage Among Smokers 

**A phase II randomized controlled tea intervention trial (4 cups/d) of decaffeinated green or black tea among smokers over a 4-mo period. **
**143 heavy smokers, aged 18-79 y, were randomized to drink either green or black tea or water. **
**Plasma and urinary levels of catechins rose significantly in the green tea group compared with the other two groups.**
**Significant decrease in urinary 8-OHdG (-31%) after 4 mo of drinking decaffeinated green tea (P = 0.002). **
**No change in urinary 8-OHdG was seen among smokers assigned to the black tea group. **

**Hakim IA, et al*****. J Nutr. 2003; 133:3303S-3309S.***

## Tea and Cancer Prevention

***Camellia Sinensis***

**Yang, CS. Personal Communication**

- Skin

- Liver

- Colon

- Oral

- Esophagus

- Lung

- Stomach

- Prostate

- Mammary

## Results of Animal Studies With Tea

| Organ Site | Protective | Not Protective |
| --- | --- | --- |
| Skin | 13 | 5 |
| Oral | 2 | 0 |
| Esophagus | 3 | 1 |
| (Fore)stomach | 5 | 0 |
| Intestine/Colon | 10 | 3 |
| Liver | 7 | 0 |
| Bladder  | 1 | 0 |
| Prostate | 1 | 0 |
| Breast | 6 | 3 |
| Lung | 10 | 1 |

**	****	****	****  ****Number of Studies**

**Yang CS. Personal Communication**

## Lung Tumorigenesis Model in A/J Mice

**Yang CS. Personal Communication**

*Tea given during the initiation stage*

- -2

- 0

- 16 weeks

- tea

- water

- 1

- NNK

*Tea given during the post-initiation stage*

- water

- tea

- 0

- 16 weeks

- 1

- NNK

*Tea given after lung adenoma formation*

- water

- tea

- 0

- 16 weeks

- 52 weeks

- NNK

- -2

****** ****Tumors**

****** ****Tumors**

****** ****Tumors**

## The TRAMP Mouse

- Transgenic Adenocarcinoma of Mouse Prostate (TRAMP) animal model
- that express the oncogene SV40 T antigen specifically in the epithelium of
- the prostate.

- Inhibition of prostate carcinogenesis in TRAMP mice by oral infusion of
- green tea polyphenols. Gupta et al. PNAS 2001;98:10350-10355.

- 100

- 50

- 0

- 30  Weeks

- 24

- 18

- 12

- 6

- metastasis

- neoplasia

- hyperplasia

- puberty

- TRAMP: A Model for Prostate Cancer Progression

- Greenberg et al. (Found on TRAMP webpage)

## Reactive Oxygen Species

**Reactive Oxygen Species**

- Endoplasmic
- Reticulum

- Mitochondrion

**O****2**

**H****2****0****2**

**OH**

-     Damage DNA,
-        RNA
-     Oxidize Proteins
-       (enzymes, histones)
-      Oxidize Lipids
-      Activate Cell
-        Suicide

- TEA

**X**

## Biological Activities of Tea Polyphenols

**Biological Activities of Tea Polyphenols**

- Induce Phase I and Phase II enzymes
- Inhibit cell proliferation and induce apoptosis
- Several effects on cell signaling pathways (e.g., cyclooxygenase)
- Inhibit angiogenesis and invasion
- Inhibit DNA methyltransferase activity

## Activation and Roles of NF-B in Oncogenesis

**NF-********B**

**Metastasis**
- (MMPs, cell adhesion molecules,
- cell surface proteases)

**Angiogenesis**
- (Cox-2, NOS, VEGF)

**Proliferation**
- (c-*myc*, cyclin D1, gro
-  cytokines [IL-2, -6])

**Oncoproteins (Her-2/neu, Ras) **

_**IKK activation**_** **
**CK2 induction**** **

**I********B**

**I********B**

**Activation and Roles of NF-********B in Oncogenesis**

**Altered cytokine production**

**NF-********B**

***nfkb***** gene amplif., rearrangement**

**Survival**
- (Bcl-xL, A1/Bfl-1,
- IEX-1L, IAP1, IAP2)

**Mutations in I********B genes**

**Co-acting factors**
**(AP-1, AhR, **
**c/EBP, p300)**

**Modifications **
**(phosphorylation, acetylation)**

**EGCG**

**[Proteasome]**

**DEX**

**Sonenshein GE.** **Personal Communication**

## Tea Polyphenol Epigallocatechin-3-Gallate Inhibits DNA Methyltransferase Activity

- Cancer Res. 63(22): 2003

## Microarray Analysis to Identify Genes Responding to EGCG Treatment in Breast Cancer Cells

**Human chip microarray (7,500 genes developed by GenomicTree, Inc, Korea)**
**Dose: 60 ********g/ml EGCG (dissolved in DMSO) or equivalent volume of DMSO, as control**
**Time points: 2, 7, and 24 hrs**
**Experiment performed twice, each time in duplicate**

**Microarray Analysis to Identify Genes Responding to EGCG Treatment in Breast Cancer Cells**

**Sonenshein GE.** **Personal Communication**

## Identification of Genes Affected by EGCG Treatment in D3-1 Breast Cancer Cells

**Identification of Genes Affected by EGCG Treatment in D3-1 Breast Cancer Cells**

**24 h samples**

**Sonenshein GE.** **Personal Communication**

## RT-PCR and Northern Blot Analysis Confirms Affects of EGCG 

**CSE-1 (****N69204**** )***

**CTGF (****AA598794****)***

**Heat shock protein 10kD (AA448396) **

**Prefoldin (AI682392)**

**ECTS (****AI031571****)***

**DMSO**

**EGCG 24 h**

**TGF******** (R36467)**

**BMP6 (AA424833)**

**GST (A4(aa152374))**

**Angiopoietin**** (T54298)***

**PTP3 (****AA995560****)***

**Thrombospondin (****AA464532****)***

**WISP-1 (AI473336)**

**GAPDH**

**Ring finger protein (AA402960)**

**AhR (AA181307)**

**28S**

**18S**

**RT-PCR**

**Northern**

**RT-PCR and Northern Blot Analysis Confirms Affects of EGCG **
**on Expression of Several Genes**

_**Genes Regulated by 24 h EGCG**_
**BMP6****        ****AhR**
**WISP-1****     ****CTGF**
**PPAR********      ****ECTS**
**GST****          ****HSP10**

**Sonenshein GE.** **Personal Communication**

## Summary

**Summary**

- Green, black and oolong teas are differentiated by tea manufacturing processes
- Teas are rich in polyphenols, e.g., catechins, theaflavins, thearubigens
- Epidemiological studies show inconsistent evidence for the association between tea consumption and reduced risk of cancer
- Preclinically, tea frequently inhibits colon, esophageal, liver, lung and skin tumorigenesis
- Tea and tea polyphenols may have multiple sites of action