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ns脉冲电场独特的诱导肿瘤细胞凋亡的生物电效应,引起了相关学者广泛的关注。为此,结合最新研究成果,侧重于ns脉冲对细胞膜结构和功能的影响,重点研究ns脉冲电场诱导肿瘤细胞凋亡的死亡受体途径。将优化的脉冲电场参数组合(电压幅值为9kV,脉宽为100ns,脉冲为30个,频率为1Hz)作用于人卵巢浆液性囊腺癌细胞SKOV3。利用流式细胞术和凝胶电泳法检测细胞凋亡、坏死情况;逆转录聚合酶链式扩增反应(reverse transcription-polymerase chain reaction,RT-PCR)法检测Fas、FasL、半胱氨酸天冬氨酸蛋白酶-8(cysteine aspartic acid specific protease-8,Caspase-8)和Bid的信使核糖核酸(messager ribonucleic acid,mRNA)释放水平;蛋白质印迹(western blot)法检测Fas、FasL、Caspase-8和Bid蛋白释放水平。实验结果表明:ns脉冲诱导SKOV3凋亡时引起了Fas、FasL、Caspase-8和Bid等释放水平的升高,从而诱导死亡受体凋亡信号通路。该研究结果大大拓展了ns脉冲诱导凋亡的机制研究,从而为临床肿瘤治疗提供理论支撑。
The unique bioelectric effect of ns pulsed electric field to induce tumor cell apoptosis has aroused widespread concern. To this end, combined with the latest research results, focusing on the ns pulse on the cell membrane structure and function, focusing on the ns pulse electric field induced death of tumor cell death receptor pathway. The optimized combination of pulsed electric field parameters (voltage amplitude 9 kV, pulse width 100 ns, pulse 30, frequency 1 Hz) was applied to human ovarian serous cystadenocarcinoma SKOV3 cells. The apoptosis and necrosis were detected by flow cytometry and gel electrophoresis. The expressions of Fas, FasL, cysteine were detected by reverse transcription-polymerase chain reaction (RT-PCR) Caspase-8 and Bid mRNA ribonucleic acid (mRNA) release levels were detected by Western blot. Fas, FasL, Caspase-8 And Bid protein release levels. The experimental results show that: ns pulse induced apoptosis of SKOV3 caused by the release of Fas, FasL, Caspase-8 and Bid and so on, thereby inducing apoptotic signal transduction death receptors. The results of this study greatly expanded the mechanism of ns pulse-induced apoptosis, thus providing theoretical support for the clinical treatment of cancer.