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Green tea constituent epigallocatechin-3-gallate and induction of apoptosis and cell cycle arrest in human carcinoma cells.

J Natl Cancer Inst 1997 Dec 17;89(24):1881-6    (ISSN: 0027-8874)

Ahmad N; Feyes DK; Nieminen AL; Agarwal R; Mukhtar H
Department of Dermatology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106, USA.

BACKGROUND AND PURPOSE: The polyphenolic compounds present in green tea show cancer chemopreventive effects in many animal tumor models. Epidemiologic studies have also suggested that green tea consumption might be effective in the prevention of certain human cancers. We investigated the effect of green tea polyphenols and the major constituent, epigallocatechin - 3 - gallate , on the induction of apoptosis ( programmed cell death) and regulation of cell cycle in human and mouse carcinoma cells. METHODS: Human epidermoid carcinoma cells ( cell line A431), human carcinoma keratinocyte ( cell line HaCaT), human prostate carcinoma cells ( cell line DU145), mouse lymphoma cells ( cell line L5178Y), and normal human epidermal keratinocytes (NHEKs) were used. Apoptosis was assessed by 1) the formation of internucleosomal DNA fragments by agarose gel electrophoresis, 2) confocal microscopy, and 3 ) flow cytometry after tagging the DNA fragments by fluorescence label. The distribution of cells in different phases of the cell cycle was analyzed by flow cytometry. RESULTS: Treatment of A431 cells with green tea polyphenols and its components, epigallocatechin - 3 - gallate , epigallocatechin , and epicatechin- 3 - gallate , resulted in the formation of internucleosomal DNA fragments, characteristic of apoptosis . Treatment with epigallocatechin - 3 - gallate also resulted in apoptosis in HaCaT, L5178Y, and DU145 cells, but not in NHEK. Confocal microscopy and flow cytometry confirmed the findings. The DNA cell cycle analysis showed that in A431 cells, epigallocatechin - 3 - gallate treatment resulted in arrest in the G0-G1 phase of the cell cycle and a dose-dependent apoptosis . CONCLUSIONS: Green tea may protect against cancer by causing cell cycle arrest and inducing apoptosis . It needs to be evaluated in human trials.