将纳秒级电场脉冲作用于人卵巢癌(SKOV3)细胞,通过流式细胞仪检测细胞活性氧(ROS)、线粒体跨膜电位(MTMP)的变化情况,用免疫荧光法检测线粒体膜间隙凋亡相关蛋白释放情况;用Western blot法检测线粒体膜间隙凋亡相关蛋白表达及活化情况,以研究纳秒级电场脉冲诱导肿瘤细胞凋亡的线粒体通路。实验发现:经纳秒级电场脉冲处理后,细胞活性氧即刻升高,且线粒体跨膜电位在6 h时达到最低值;线粒体膜间隙蛋白释放入胞浆(P〈0.05);线粒体凋亡通路相关蛋白表达量也明显升高(P〈0.05)。结果表明,在纳秒级电场脉冲诱导的肿瘤细胞凋亡中,线粒体凋亡通路参与并发挥了重要作用。
In order to study the mitochondrial signaling pathways of tumor cell apoptosis induced by nanosecond pulsed electric fields(nsPEF),nsPEF with specific parameters were performed on SKOV3 cells,then changes of reactive oxygen species(ROS) mitochondrial trans-membrane potential(MTMP) were tested by flow cytometry;releases of mitochondrial membrane space apoptosis-related proteins were studied by immunofluorescence;expressions or activations of mitochondrial membrane space apoptosis-related proteins were tested by Western blot.The experimental results showed that ROS increased instantly,and MTMP reduced to the minimum value 6 h after nsPEF;mitochondrial membrane space apoptosis-related proteins released to cytoplasm(P 0.05),and expressions of these proteins were higher than the control group(P 0.05).The results indicate that mitochondrial apoptosis signaling pathway exists and plays an important role in tumor cell apoptosis with nsPEF.