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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。
Objective: To study the apoptosis of human cervical carcinoma HeLa cells induced by Vitexin and to explore the mechanism of apoptosis induced by Vitexin. Methods: Human cervical cancer HeLa cells were cultured in vitro. The apoptotic ELISA kit was used to detect the histone / DNA fragments of HeLa cells. The percentage of Sub-G1 cells was quantified by propidium iodide staining and the DNA ladder was observed by DNA agarose gel electrophoresis. caspase colorimetric kit was used to detect the activity of caspase-3 / -9; Mitochondrial membrane potential was detected by Rh123 probe staining and the expression of cytochrome c, Bax, Bcl-2, Bcl-xL and XIAP was detected by Western blot. Results: After treatment with different concentrations of CAS for 48h, the number of Histone / DNA fragments and the percentage of Sub-G1 cells in HeLa cells increased significantly. DNA ladder was observed by DNA agarose gel electrophoresis. Caspase-3 / -9 activity, mitochondrial membrane potential decreased, cytochrome c, Bax increased significantly, while Bcl-xL and XIAP protein levels decreased. Conclusion: CAS induces HeLa cell apoptosis through mitochondrial apoptosis pathway.