表没食子儿茶素没食子酸酯对人卵巢癌细胞株SKOV3自噬的影响

Effects of EGCG on the autophagy of human ovarian cancer SKOV3 cells

目的：探讨表没食子儿茶素没食子酸酯（ EGCG）对人卵巢癌株SKOV3自噬的影响。方法体外培养SKOV3细胞，不同浓度EGCG作用于SKOV3细胞不同时间后，采用MTT比色法检测SKOV3细胞增殖的情况；选取低、中、高浓度的EGCG处理SKOV3细胞24 h，用实时荧光定量PCR法（ real－time FQ－PCR）检测Beclin1 mR-NA表达水平，用Western blot法检测Beclin1蛋白的表达水平。结果 MTT比色法检测显示，与对照组细胞比较，上述不同浓度的EGCG作用不同时间后，SKOV3细胞增殖抑制率明显增加（P＜0．05），且随着EGCG浓度的增加及作用时间的延长，其抑制作用呈明显的计量和时间依赖性（P＜0．05）。 Real－time FQ－PCR检测显示，不同浓度EGCG作用后，SKOV3细胞中 Beclin1 mRNA 表达水平与相应对照组细胞比较差异均有统计学意义（ P＜0．05）；Western blot显示，随着EGCG浓度的增加，SKOV3细胞中Beclin1蛋白表达水平逐渐下降，其作用浓度为中与高浓度EGCG组分别与对照组细胞比较，差异均有统计学意义（P＜0．05）。结论 EGCG对SKOV3细胞具有增殖抑制作用，同时可诱导SKOV3细胞自噬水平降低，提高患者药物治疗效果，值得临床应用。

Objective To investigate the effects of epigallocatechin -3-gallate （ EGCG） on the autophagy of hu-man ovarian cancer SKOV 3 cells.Methods SKOV3 cells were cultivated in vitro and exposed to different concentrations of EGCG for different times .Then, their proliferation was determined by MTT assay .The cells were also treated with dif-ferent concentrations of EGCG for 24 hours and the levels of autophagy -related genes Beclin1 mRNA and protein were measured by real-time quantitative fluorescent PCR （ real-time FQ-PCR） and Western blotting .Results According to MTT assay, exposure to EGCG could remarkably inhibit the proliferation rate of SKOV 3 cells in a dose-and time-dependent manner in comparison with the control （P〈0.05）.Real-time FQ-PCR results showed that, the level of Beclin1 mRNA in SKOV3 cells exposed to different concentrations of EGCG was significantly different from that in the control （P〈0.05）.Western blotting analysis indicated that , the level of Beclin1 was gradually down -regulated along the increase of EGCG concentrations , and statistical difference could be found at a medium and high concentration of EGCG （P〈0.05）.Conclusion EGCG can inhibit the proliferation of SKOV3 cells, and suppress their autophagy, which improves therapeutic efficacy and is worthy of clinical application .