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Objective: To observe the table epigallocatechin gallate (Epigallaocatechin-3-gallate, EGCG) , and to explore the molecular mechanisms of human colon cancer HT-29 cell proliferation . METHODS: Cultured human colon cancer HT-29 cells , using MTT colorimetric detection of EGCG on HT-29 cell growth inhibition ; using flow cytometry analysis of EGCG on HT-29 cell cycle distribution ; using immunocytochemistry the immunoblotting methods such as observation EGCG expression in HT-29 cell cycle-related protein (p53, PCNA, p38MAPK, RB) . RESULTS: MTT colorimetric results show that EGCG can in a concentration -dependent inhibition of proliferation of HT-29 cells . Different concentrations of EGCG (30μg/ml, 40μg/ml, 50μg/ml, 60μg/ml) significant growth inhibitory effect of HT-29 cells (24h, 48h, 72h, 96h) and dose - and time - effect dependence (P lt; 0.05); cell viability test results showed that the control group and the EGCG -treated cells of various concentrations of no significant difference in survival sex ( P gt ; 0.05) ; under the light microscope , compared with the control group , treatment group tumor cells sparse , the number of significantly reduced cell atypia smaller , more consistent morphology , abundant cytoplasm , nuclear becomes smaller , more consistent size , a larger proportion of nuclear pulp obvious nucleoli, abundant cytoplasmic organelles ; flow cytometry the analysis results show that the induction of EGCG in human colon cancer cells in G1 phase arrest , and with the treatment time , the more obvious effects of the induction cycle arrest ( P lt; 0.05 ) , the same time treated with different concentrations dealing with the blank according occurred G1 arrest (P lt; 0.05); immunocytochemistry quantitative image analysis : EGCG ( 50μg/mL , 48h) after treatment of human colon cancer HT-29 cells , p53, PCNA protein positive rate than the untreated group was significantly lower (P lt; 0.05); Western blot : EGCG can cut of p38MAPK, RB protein phosphorylation level , its role in the concentration and time , can down-regulate the expression of Cyclin D1 protein . Conclusion : 1.EGCG cultured human colon cancer HT-29 cells with significant inhibition of proliferation and a dose - and time-dependent relationship . 2.EGCG inhibit HT-29 cell proliferation and induced cell cycle G1 phase arrest. 3.EGCG possibly through inhibition of p38MAPK phosphorylation of RB protein down of Cyclin D1 , p53 , PCNA protein expression induced by HT-29 cells in G1 phase arrest , inhibition of proliferation of HT-29 cells .
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