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目的:探讨表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate,EGCG)诱导T24细胞凋亡及对Survivin表达的调控作用。方法:不同剂量(0~80μg/mL)表没食子儿茶素没食子酸酯与T24细胞同培养不同时间(12、24、48 h),采用CCK8法测定细胞的增殖,流式细胞术测定细胞凋亡率。用不同剂量表没食子儿茶素没食子酸酯与细胞培养24 h后,采用Western blot测定Survivin蛋白的表达情况,用半定量反转录聚合酶链式法(Reverse Transcription-Polymerase Chain Reaction,RT-PCR)测定Survivin mRNA表达。结果:不同浓度EGCG处理T24细胞,呈浓度及时间依赖性抑制T24细胞增殖;流式细胞仪检测发现EGCG作用T24细胞24 h后,细胞出现凋亡。Western blot及半定量RT-PCR检测细胞中Survivin及其mRNA的表达,结果显示EGCG可下调Survivin表达。结论:EGCG能抑制T24细胞增殖,诱导其凋亡,其机制可能与下调细胞中Survivin表达相关。
Objective: To investigate the effect of epigallocatechin-3-gallate (EGCG) on the apoptosis of T24 cells and the regulation of Survivin expression. Methods: Different concentrations of epigallocatechin gallate (0 ~ 80μg / mL) were incubated with T24 cells for different times (12,24,48 h). Cell proliferation was measured by CCK8 assay. Flow cytometry Death rate. Survivin protein expression was determined by Western blotting with different doses of epigallocatechin-gallate and cell culture for 24 h, and the expression of Survivin protein was detected by reverse transcriptase-Polymerase Chain Reaction (RT-PCR) ) Survivin mRNA expression was measured. Results: T24 cells treated with different concentrations of EGCG inhibited the proliferation of T24 cells in a concentration-and time-dependent manner. The apoptosis of T24 cells induced by EGCG was observed by flow cytometry. Western blot and semi-quantitative RT-PCR assay Survivin and its mRNA expression, the results showed that EGCG down-regulated Survivin expression. Conclusion: EGCG can inhibit the proliferation of T24 cells and induce its apoptosis. The mechanism may be related to the down-regulation of Survivin expression.