表没食子儿茶素没食子酸酯对人宫颈癌细胞系HeLa细胞凋亡及bcl―2bax表达影响.docVIP

表没食子儿茶素没食子酸酯对人宫颈癌细胞系HeLa细胞凋亡及bcl―2bax表达影响 　　[摘要] 目的 探讨表没食子儿茶素没食子酸酯（EGCG）对宫颈癌细胞系HeLa细胞凋亡及bcl-2/bax表达的影响。 方法 体外培养HeLa细胞，分为对照组和实验组，实验组用不同浓度的EGCG处理24、48、72 h，对照组加入等体积的培养基。采用MTT法测定EGCG对HeLa细胞增殖的影响；倒置显微镜下观察细胞的形态学变化；EGCG（浓度分别为20、40、80 μg/mL）处理HeLa细胞48 h后，分别采用流式细胞仪检测细胞凋亡和周期、分光光度计检测半胱氨酸天冬氨酸蛋白酶（Casepase-3）相对活性、Western blot法检测bcl-2/bax蛋白表达情况。 结果 EGCG（浓度10～100 μg/mL）能抑制HeLa细胞的增殖，呈剂量和时间依赖性；倒置显微镜下可观察到典型的细胞凋亡形态；浓度为20、40、80 μg/mL的EGCG作用HeLa细胞48 h后，细胞凋亡率逐渐上升，分别为12.4%、23.4%、35.4%，明显高于对照组（3.3%）；细胞周期结果显示EGCG能抑制细胞的G1期行进和G1/S转换，降低S期细胞比例；实验组HeLa细胞Caspase-3相对活性分别为（1.36±0.07）、（1.85±0.06）、（2.45±0.07），均高于对照组（0.93±0.06），差异均有统计学意义（P 0.05）；bax蛋白表达量升高，而bcl-2蛋白表达量降低，呈剂量依赖性。 结论 EGCG能抑制HeLa细胞的增殖并诱导其凋亡，其作用机制可能与增加细胞Casepase-3活性、下调bcl-2基因的表达及上调bax基因有关。EGCG是一种前景广阔的抗肿瘤药物。 　　[关键词] 表没食子儿茶素没食子酸酯；HeLa；半胱氨酸天冬氨酸蛋白酶 　　[中图分类号] R285 [文献标识码] A [文章编号] 1673-7210（2014）08（c）-0029-05 　　Effects of EGCG on cell apoptosis in cervical cancer HeLa cells and expression of bcl-2/bax 　　QIN Jing HONG Li 　　The Second Department of Gynecology， Renmin Hospital of Wuhan University， Hubei Province， Wuhan 430060， China 　　[Abstract] Objective To explore the effect of EGCG on cell apoptosis in cervical cancer line HeLa cells and expression of bcl-2/bax. Methods HeLa cells were cultured in vitro， and they were divided into two groups， including control group and experimental group. The control group was received only the culture medium， and the experimental group was treated by EGCG at different concentrations for 24、48、72 h respectively. The cell survival of the two groups were determined by the MTT method； the changes of cell morphology were observed by inverted microscope； apoptosis and cell cycle were analyzed by flow cytometry； the relative activity of caspase-3 was monitored by spectrophotometer； the changes of protein expression of bcl-2 and bax were detected by Western blot. Results From the data of MTT， the cell proliferation of human cervical cancer HeLa cells was inhibited by EGCG （10-100 μg/mL） in a dose-dependent a