摘要
目的:探讨七叶皂苷钠诱导宫颈癌HeLa细胞凋亡的作用及其分子机制。方法:采用MTT法检测七叶皂苷钠对宫颈癌HeLa细胞的生长和增殖抑制作用;倒置显微镜观察细胞形态改变;利用annexin V-FITC/PI流式细胞术检测细胞凋亡率;采用DAPI单染法荧光显微镜下观察细胞核变化情况;利用Western blotting检测凋亡相关蛋白[聚(ADP-核糖)聚合酶(PARP)、cleaved caspase-8、cleaved caspase-9、pro-caspase-3]和细胞存活相关信号通路(Akt、ERK)以及TRAIL受体(DR4、DR5)的变化情况。结果:七叶皂苷钠以剂量依赖的方式显著抑制宫颈癌HeLa细胞的生长和增殖;七叶皂苷钠作用于HeLa细胞后,可见典型的凋亡细胞形态学特征,细胞凋亡率显著增加;随着七叶皂苷钠浓度升高,cleaved PARP、cleaved caspase-8和cleaved caspase-9明显增多,pro-caspase-3显著减少,p-Akt和p-ERK激活减少,细胞内DR4和DR5总蛋白水平上调。结论:七叶皂苷钠通过抑制细胞存活相关信号通路,上调死亡受体水平,抑制细胞增殖,促进细胞凋亡。
AIM: To study the effects of sodium aescinate on the apoptosis of cervical cancer HeLa cells and its molecular mechanism. METHODS : MTT assay was used to detect the growth and proliferation of HeLa cells. The mor- phological alteration was observed under inverted microscope. Annexin V-FITC/PI double staining and DAPI nuclear stai- ning were used to determine the apoptosis of HeLa cells induced by sodium aescinate. The apoptosis-related proteins PARP, cleaved caspase-8 and pro-caspase-3, and the proliferation-associated molecules Akt and ERK, as well as TRAIL receptors DR4 and DR5 were detected by Western blotting. RESULTS: Sodium aescinate inhibited the growth of HeLa ceils in a concentration-dependent manner. Treatment with sodium aescinate induced the typical morphology of apoptotic cells and increased the apoptotic rate significantly. The cleaved PARP, cleaved caspase-8 and cleaved caspase-9 protein expression was observed. The expression of DR4 and DR5 was up-regulated. Meanwhile, pro-caspase-3 was decreased, and the levels of p-Akt and p-ERK were down-regulated by sodium aescinate in a dose-dependent manner. CONCLU- SION : Sodium aescinate inhibits the proliferation and promotes the apoptosis of HeLa cells by increasing death receptor ex- pression and repressing proliferation-associated signaling pathways.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2014年第2期239-244,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.31301171)
活性生物大分子研究安徽省重点实验室项目(No.1306C083008)
高等学校省级优秀青年人才基金重点项目(No.2013SQRL055ZD)
皖南医学院博士科研启动基金资助项目