Involvement of protein tyrosine phosphorylation in the effect of green tea polyphenols on Ehrlich ascites tumor cells in vitro.
Green tea extract and its polyphenolic components have been found to possess anticarcinogenic, antimutagenic, antihypertensive and antihepatotoxic effects, and several mechanisms have been proposed for these effects. In this study, the effects of five tea polyphenols, (-)-epigallocatechin-3-gallate (EGCG), (-)-epigallocatechin (EGC), (-) epicatechin-3-gallate (ECG), ( -) epicatechin (EC) and (+)-catechin (C), were examined on the viability of Ehrlich ascites tumor cells in vitro and a possible relationship with tyrosine phosphorylation was determined. Proteins extracted from the cells treated with the tea polyphenols were separated by SDS-PAGE, and tyrosine-phosphorylated proteins were detected by immunoblotting with anti-phosphotyrosine antibody and the extent of phosphorylation determined. EGC (100 microM) caused a significant decrease in cell viability to 4.1 +/- 0.2% of the control value, and this correlated with a stimulation of protein tyrosine kinase (PTK) activity. EGCG (100 microM) also caused a slight decrease in cell viability (approximately 70% of the control value) but this and the other polyphenols, which had no effect on cell viability likewise, had no effect on tyrosine phosphorylation. Tyrosine phosphorylations of 42 and 45 kDa proteins were also observed for EGC. Further evaluation of the effect of EGC showed that the activity of ornithine decarboxylase (ODC), a key enzyme in polyamine biosynthesis in cells, decreased significantly as well. A significant correlation has therefore been observed between a cellular event, namely, a reduction in the viability of Ehrlich ascites tumor cells and an association with a tyrosine phosphorylation of 42 and 45 kDa proteins by the polyphenol EGC.
Kennedy
D O
DO
Department of Food and Nutrition, Faculty of Human Life Science, Osaka City University, Osaka, Japan.
Nishimura
S
S
Hasuma
T
T
Yano
Y
Y
Otani
S
S
Matsui-Yuasa
I
I
D000818:Animals; D016588:Anticarcinogenic Agents / Q000494:pharmacology*; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism*; D002392:Catechin / Q000031:analogs & derivatives / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D005419:Flavonoids / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D009363:Neoplasm Proteins / Q000378:metabolism*; D009955:Ornithine Decarboxylase / Q000378:metabolism; D010636:Phenols / Q000494:pharmacology*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D013662:Tea; D014407:Tumor Cells, Cultured
D016588:Anticarcinogenic Agents; D005419:Flavonoids; D009363:Neoplasm Proteins; D010636:Phenols; D013662:Tea; D019000:Phosphotyrosine; D002392:Catechin; C062669:epicatechin gallate; C045651:epigallocatechin gallate; D011505:Protein-Tyrosine Kinases; D009955:Ornithine Decarboxylase; C057580:gallocatechol