去甲斑蝥素诱导超氧阴离子的产生介导HeLa细胞线粒体途径的细胞凋亡

Eff ect of the norcantharidin-induced production of superoxide anion on mediating the apoptosis of HeLa cells in the mitochondrial pathway

目的探讨去甲斑蝥素(norcantharidin,NCTD)诱导的超氧阴离子对人宫颈癌HeLa细胞凋亡的影响。方法采用流式细胞术检测细胞凋亡率和线粒体膜电位,荧光显微镜下观察罗丹明123染色后线粒体膜电位的变化,蛋白印迹法检测线粒体途径细胞凋亡相关蛋白的表达。结果60μmol/L去甲斑蝥素作用24 h后增加了细胞凋亡率,诱导了线粒体膜电位的丧失,降低Bcl-2蛋白表达,增加Bax蛋白表达。5 mmol/L超氧阴离子清除剂超氧化物歧化酶(SOD)作用后逆转了去甲斑蝥素诱导的细胞凋亡率和线粒体膜电位的崩溃,同时还逆转了去甲斑蝥素诱导的Bax表达增加和Bcl-2的表达水平降低。结论去甲斑蝥素诱导超氧阴离子的产生介导了线粒体途径的细胞凋亡。

Objective To investigate the eff ect of the superoxide anion induced by norcantharidin(NCTD)on the apoptosis of HeLa cells in human cervical cancer.Methods The cell apoptosis rate and mitochondrial membrane potential were detected by the flow cytometry.The changes of the mitochondrial membrane potential after the rhodamine 123 staining were observed under a fl uorescence microscope,and the expression of the apoptosis-related proteins in the mitochondrial pathway was detected by Western blotting.Results After being treated with the 60μmol/L NCTD for 24 h,the rate of apoptosis was increased,the loss of mitochondrial membrane potential was induced,the expression of Bcl-2 protein was decreased,and the expression of Bax protein was increased.After being treated with the 5 mmol/L superoxide anion scavenger superoxide dismutase(SOD),the apoptosis rate and mitochondrial membrane potential collapse induced by NCTD were reversed.Meanwhile,the increase of Bax expression and the decrease of Bcl-2 expression induced by NCTD were also reversed.Conclusion The NCTD-induced production of superoxide anions mediates the apoptosis in the mitochondrial pathway.