紫花牡荆素通过线粒体凋亡途径诱导人宫颈癌HeLa细胞凋亡

目的:研究紫花牡荆素诱导人宫颈癌HeLa细胞凋亡作用,并探讨其诱导凋亡机制.方法:体外培养人宫颈癌HeLa细胞.细胞凋亡ELISA试剂盒检测HeLa细胞组蛋白/DNA碎片；碘化丙啶染色流式细胞术定量分析Sub-G1细胞百分率；DNA琼脂糖凝胶电泳观察DNA梯形条带.caspase比色试剂盒检测caspase-3/-9活性；Rh123探针染色FCM检测线粒体膜电位;Western blot检测细胞色素c、Bax、Bcl-2、Bcl-xL和XIAP蛋白的表达.结果:不同浓度的CAS处理48h后,HeLa细胞组蛋白/DNA碎片增加、Sub-G1细胞百分率显著增高、DNA琼脂糖凝胶电泳法观察到典型的DNA梯形条带;CAS处理组caspase-3/-9活性明显升高、线粒体膜电位降低、细胞色素c、Bax显著升高,而Bcl-xL和XIAP蛋白表达水平降低.结论:CAS通过线粒体凋亡途径诱导HeLa细胞凋亡.

Induction of apoptosis by casticin in cervical cancer cells through mitochondrial pathway

Objective The purpose of this study was to examine the apoptotic activity of casticin on human cervical cancer HeLa cell and its molecular mechanism.Methods Apoptotic activities of casticin on human cervical cancer HeLa were performed using histone/DNA ELISA assay,flow cytometry with propidium iodide(PI) staining and DNA agarose gel electrophoresis.Caspase activities were assayed using caspase colorimetric activity assay kit.The mitochondrial membrane potential was evaluated by flow cytometry analysis after Rh123 probe staining.Protein expression of cytochrome c,Bax,Bcl-2,Bcl-xL and XIAP was performed by Western blot.Results Casticin caused histone/DNA increased,the Sub-G1 population accumulation and the typical DNA ladder in HeLa cell.The colorimetric analysis showed the activation of caspase-3 and-9 induced by casticin.Mitochondrial transmembrane potential was reduced,Bax and cytochrome c was up regulated and there was down regulation of Bcl-xL and XIAP expression after treatment with casticin.However,the expression of Bcl-2 showed no change in such same treatment.Conclusion These results indicate that casticin induced apoptois of cervical cancer HeLa cells is mediated by mitochondrial pathway.