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目的:观察表没食子儿茶素没食子酸酯(Epigallaocatechin-3-gallate,EGCG)对人结肠癌HT-29细胞增殖和凋亡的影响,并探讨其对MMP-2,RECK的调节作用。方法:体外培养人结肠癌HT-29细胞,MTT比色法检测EGCG对HT-29细胞的生长抑制作用;Histone/DNA ELISA检测细胞凋亡;FITC标记Annexin-V/PI双染流式细胞术分析凋亡细胞百分率;Western Blot和RT-PCR方法检测EGCG对MMP-2,RECK蛋白和mRNA表达的影响。结果:EGCG呈浓度和时间依赖性抑制HT-29细胞的增殖,并且增加HT-29细胞Histone/DNA碎片的渗漏;EGCG诱导HT-29细胞凋亡百分率增高;EGCG抑制MMP-2蛋白和mRNA的表达,促进RECK蛋白和mRNA的表达。结论:EGCG抑制人结肠癌HT-29细胞的增殖,促进其凋亡,并且呈浓度和时间依赖性;其作用机制可能与其下调MMP-2蛋白和mRNA的表达、上调RECK蛋白和mRNA的表达有关。
OBJECTIVE: To observe the effect of epigallaocatechin-3-gallate (EGCG) on the proliferation and apoptosis of human colon cancer HT-29 cells and to explore the regulation of MMP-2 and RECK. Methods: The human colon cancer HT-29 cells were cultured in vitro. The growth inhibition of HT-29 cells was detected by MTT assay. The apoptosis was detected by Histone / DNA ELISA. FITC-labeled Annexin-V / PI double staining flow cytometry The percentage of apoptotic cells was analyzed. The effects of EGCG on the expression of MMP-2, RECK protein and mRNA were detected by Western Blot and RT-PCR. Results: EGCG inhibited the proliferation of HT-29 cells in a concentration-and time-dependent manner and increased the leakage of Histone / DNA fragments in HT-29 cells. The percentage of apoptosis induced by EGCG increased in HT-29 cells. The expression of RECK protein and mRNA is promoted. CONCLUSION: EGCG can inhibit the proliferation and promote the apoptosis of human colon cancer HT-29 cells in a concentration-and time-dependent manner. The mechanism may be related to down-regulation of MMP-2 protein and mRNA expression and up-regulation of RECK protein and mRNA expression .