乳浆大戟提取物诱导HeLa细胞凋亡及其机制研究

Study on the mechanism of Euphorbia esula extract induced HeLa cells apoptosis

目的研究乳浆大戟提取物诱导宫颈癌HeLa细胞凋亡的作用及其分子机制。方法采用MTT法和克隆形成实验检测乳浆大戟提取物对宫颈癌HeLa细胞的生长和增殖抑制作用;利用Annexin V-FITC/PI双染和DAPI细胞核染色法检测乳浆大戟提取物诱导细胞凋亡情况;线粒体膜电位检测凋亡细胞的线粒体膜电位变化情况;流式细胞仪和Westernblot法分别分析细胞周期及细胞周期蛋白变化情况。结果①乳浆大戟提取物以时间和剂量依赖的方式显著抑制宫颈癌HeLa细胞的生长和增殖;②乳浆大戟提取物能使宫颈癌HeLa细胞线粒体膜电位显著降低,诱导细胞发生凋亡性细胞死亡;③乳浆大戟提取物能够显著阻滞宫颈癌HeLa细胞在G0/G1期,降低Cyclin D1、Cyclin E和CDK4的表达水平,增加P21的表达水平。结论乳浆大戟提取物通过下调Cyclin D1、Cyclin E和CDK4的表达和上调P21的表达,使HeLa细胞阻滞在G0/G1期,进而显著抑制宫颈癌HeLa细胞生长和增殖,诱导细胞依赖线粒体通路的凋亡性死亡。

Objective To study the impact of Euphorbia esula extract induced HeLa cells of cervical apoptosis and its molecular mechanism.Methods MTT assay and clonogenic survival assay were used to detect the growth and proliferation of cervical cells;Annexin V-FITC/PI double staining and DAPI nuclear staining were used to detect the apoptosis which were induced by Euphorbia esula extract;Mitochondrial membrane potential change was investigated with JC-1 assay;Cell cycle change and cell cycle related proteins were respectively analyzed by flow cytometry and Western blot method.Results（1） Euphorbia esula extract can significantly inhibit the growth and proliferation of HeLa cells of cervial cencer in a time and dose-dependent manner.（2） Euphorbia esula extract significantly reduced the mitochondrial membrane and induced HeLa cells of cervical cencer to die by apoptosis.（3） Euphorbia esula extract significantly arrested the cells growth in the G0/G1 phase,reduced the expression levels of Cyclin D1,Cyclin E and CDK4 proteins,and increased the expression levels of P21 protein.Conclusion Euphorbia esula extract can induce the cell cycle arrest at G0/G1 phase by down-regulating the expression levels of Cyclin D1,Cyclin E and CDK4 proteins,and up-regulating the expression levels of P21 protein.Therefore,Euphorbia esula extract can efficiently inhibit the cell proliferation and the apoptosis of HeLa cells.