【摘要】 目的:探讨纳秒级脉冲电场对SKOV3细胞的凋亡诱导作用及对凋亡相关蛋白Bcl-2/Bax表达的影响。方法:以场强为50kV/cm,脉宽100ns,10、30、60或80个脉冲电场处理SKOV3细胞,以未经处理的SKOV3细胞为对照组,Annexin V/PI双染流式细胞仪检测早期凋亡;处理后6h,琼脂糖凝胶电脉法检测晚期凋亡;以50kV/cm,100ns,30个脉冲电场处理细胞,分别孵育0h,2h,6h,12h后提取总蛋白,Western blot法检测caspase-3活性片段、Bax、Bcl-2表达量的改变。结果:与对照组相比,纳秒级陡脉冲处理组细胞出现明显的磷脂酰丝氨酸(PS)外翻(P<0.05)且细胞膜保持完整,即Annexin V(+)/PI(-);凝胶电泳检测显示,处理组DNA呈明显的阶梯状分布;Western blot检测示,与对照组相比,处理组细胞内caspase-3活性片段、促凋亡蛋白Bax表达量显著增加(P<0.05),凋亡抑制蛋白Bcl-2表达量略降低,Bcl-2/Bax显著降低(P<0.05)。结论:纳秒级脉冲电场可通过caspase-3依赖凋亡途径,降低Bcl-2/Bax比值诱导SKOV3细胞凋亡。更多还原

【Abstract】 Objective:To investigate the effect of nanosecond pulsed electric fields(nsPEF)on apoptosis and Bcl-2/Bax expression in SKOV3 cells.Methods:SKOV3 cells were exposed to the 100ns,50kV/cm,10,30,60 and 80 pulses(as the treated groups),Annexin V/PI double-stain and flow cytometry were used to assay early stage apoptosis.Agarose gel electrophoresis was used to detect DNA ladder after 6 h treated with nsPEF;Western blot was used to measure the level of cleaved caspase-3,Bcl-2 and Bax after exposing to 100ns,50kV/cm,30 pulses electric field at 0,2,6 and 12 h,respectively.Results:Compared with the control group,phosphatidylserine externalized(P<0.05),and a typical DNA ladder pattern of internucleosomal fragmentation was observed,expression of cleaved capase-3 and Bax was increased(P<0.05);Bcl-2 and Bcl-2/Bax ratio decreased(P<0.05)in nsPEF-treatment groups.Conclusion:caspase-3 dependent apoptotic pathway and the decrease in Bcl-2/Bax ratio may be involved in the apoptosis induced by the nsPEF.更多还原