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为探讨纳秒级陡脉冲(nsPEFs)对人卵巢癌SKOV3细胞凋亡作用及Fas介导的细胞凋亡通路的影响,以场强为45kV/cm,脉宽分别为50、100和200ns,脉冲个数为30个的nsPEF处理的SKOV3细胞为实验组,以未经处理的SKOV3细胞为对照组,流式细胞术检测细胞晚期凋亡率和(或)死亡率,琼脂糖凝胶电泳法检测DNA Ladder,RT-PCR和Western blot检测细胞内Fas、FasL、caspase-8和Bid的表达量变化。结果显示与对照组相比,脉宽分别为50、100和200ns组细胞晚期凋亡率和(或)死亡率分别为(18.31%±0.65%)、(45.55%±3.71%)和(47.47%±7.01%),明显高于对照组(3.03%±0.57%)(P<0.05);琼脂糖凝胶电泳结果显示脉宽为50ns和100ns组有大量DNA ladder形成,而200ns组条带模糊;RT-PCR检测结果显示50ns、100ns组Fas、FasL、caspase-8和Bid mR-NA表达量均显著升高(P<0.05),200ns组mRNA表达量均显著降低(P<0.05);Western blot检测结果显示50ns、100ns组Fas、FasL、caspase-8和Bid蛋白表达量显著升高(P<0.05),200ns组蛋白表达量均显著降低(P<0.05)。提示场强45kV/cm,脉宽为50ns、100ns的nsPEFs能诱导人卵巢癌SKOV3细胞发生凋亡,机理可能与Fas介导的细胞凋亡通路被激活有关。
To investigate the effects of nsPEFs on the apoptosis of human ovarian cancer cell line SKOV3 and Fas-mediated apoptosis, the effects of pulse voltage The number of NSPEF-treated SKOV3 cells was 30 in the experimental group. The untreated SKOV3 cells were used as the control group. The apoptosis rate and / or mortality of the cells were detected by flow cytometry. The agarose gel electrophoresis DNA Ladder, RT-PCR and Western blot were used to detect the expression of Fas, FasL, caspase-8 and Bid. The results showed that compared with the control group, the rates of late apoptosis and / or mortality in the groups of 50, 100 and 200ns pulse width were (18.31% ± 0.65%), (45.55% ± 3.71%) and (47.47% ± 7.01%) was significantly higher than that of the control group (3.03% ± 0.57%) (P <0.05). The results of agarose gel electrophoresis showed that a large number of DNA ladder formed in the pulse width of 50ns and 100ns group, The results of RT-PCR showed that the expression of Fas, FasL, caspase-8 and Bid mR-NA in 50ns and 100ns groups were significantly increased (P <0.05), and the mRNA expression in 200ns group was significantly decreased (P <0.05) The results showed that the expressions of Fas, FasL, caspase-8 and Bid in 50ns and 100ns groups were significantly increased (P <0.05), and those in 200ns group were significantly decreased (P <0.05). Prompted field strength 45kV / cm, pulse width of 50ns, 100ns nsPEFs can induce apoptosis in human ovarian cancer SKOV3 cells, the mechanism may be related to Fas-mediated apoptosis pathway is activated.