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目的:研究表没食子儿茶素没食子酸酯(Epigallaocatechin-3-gallate,EGCG)对人结肠癌HT-29细胞增殖的影响。方法:实验分为EGCG不同浓度处理组和阴性对照组,采用MTT比色法检测EGCG(30μg/mL、40μg/mL、50μg/mL、60μg/mL、70μg/mL)对HT-29细胞的生长影响;应用流式细胞术分析EGCG对HT-29细胞周期分布的影响;免疫印迹观测EGCG对HT-29细胞p38MAPK、cyclinD1蛋白表达的影响。结果:MTT比色结果显示,不同浓度EGCG(30μg/ml、40μg/ml、50μg/ml、60μg/ml)对HT-29细胞具有明显的生长抑制作用,并呈剂量-效应依赖关系(P<0.05);流式细胞术分析显示,EGCG诱导人结肠癌细胞G1期阻滞,且随着处理时间的延长,其诱导周期阻滞的效应越明显(P<0.05);蛋白免疫印迹显示,总的p38MAPK不随处理时间和浓度的改变而改变,但是磷酸化的p38MAPK蛋白的表达随处理时间和处理浓度的增加而明显增加,而CyclinD1蛋白的表达随处理浓度的增加而明显减少。结论:EGCG诱导HT-29细胞G1期阻滞,抑制细胞增殖,可能与活化p38MAPK,下调CyclinD1蛋白表达有关。
Objective: To study the effect of epigallaocatechin-3-gallate (EGCG) on the proliferation of human colon cancer HT-29 cells. Methods: The experiment was divided into different concentrations of EGCG treatment group and negative control group. MTT colorimetric method was used to detect the growth of HT-29 cells induced by EGCG (30μg / mL, 40μg / mL, 50μg / mL, 60μg / mL and 70μg / mL) The effect of EGCG on the cell cycle distribution of HT-29 cells was analyzed by flow cytometry. The effect of EGCG on the expression of p38MAPK and cyclinD1 protein in HT-29 cells was observed by immunoblotting. Results: The results of MTT assay showed that EGCG at different concentrations (30μg / ml, 40μg / ml, 50μg / ml and 60μg / ml) significantly inhibited the growth of HT-29 cells in a dose-dependent manner (P < 0.05). Flow cytometry analysis showed that EGCG induced cell cycle arrest in human colon cancer cell line G1, and the more obvious the induction of cell cycle arrest (P <0.05) with the prolongation of treatment time (P <0.05) P38MAPK did not change with the time and concentration of phosphorylation, but the expression of phosphorylated p38MAPK protein increased with the increase of treatment time and concentration, while the expression of CyclinD1 protein decreased with the increase of the concentration of p38MAPK. Conclusion: EGCG can induce G1 phase arrest and inhibit cell proliferation in HT-29 cells, which may be related to activation of p38MAPK and down-regulation of CyclinD1 protein expression.