纳秒脉冲电场诱导肿瘤细胞凋亡的内质网信号通路分析

ns脉冲电场在诱导肿瘤细胞凋亡方面展现出诱人的应用前景。由于ns脉冲电场诱导肿瘤细胞凋亡的机制尚未明确,严重阻碍了ns脉冲电场肿瘤治疗的临床进程。为进一步揭示其诱导凋亡的机制,将参数组合(电压幅值9kV,脉宽100ns,脉冲30个,频率1Hz)作用于人卵巢浆液性囊腺癌(SKOV3)细胞。首先检测了半胱氨酸天冬氨酸蛋白酶-12(cysteine aspartic acid specific protease-12,Caspase-12)蛋白的释放水平,其次用浓度分别为0、25、50、100μmol/L的钙螯合剂(BAPTA-AM)螯合细胞内的游离Ca2+,用Hoechst 33342荧光染色检测细胞凋亡率,最后采用蛋白质印迹(western blot)技术检测了Caspase-12蛋白释放的变化。试验结果表明:ns脉冲诱导SK-OV3凋亡时引起了细胞内Ca2+升高,从而介导内质网凋亡信号通路。该研究结果将进一步揭示ns脉冲诱导凋亡的机制。

Analysis of Endoplasmic Reticulum Apoptotic Signaling Pathway of Tumor Cells Induced by Nanosecond Pulsed Electric Field

Tumor cells apoptosis by using nanosecond pulsed electric field(nsPEF) has a great potential in clinical tumor treatment.The clinic progress of nsPEF in tumor treatment is seriously impeded because of unclear mechanisms of tumor cell apoptosis with nsPEF.To probe the mechanisms of nsPEF-induced tumor cell apoptosis,optimized parameters(voltage amplitude of 9 kV,pulse duration of 100 ns,pulse number of 30,repetition frequency of 1 Hz) of nsPEF were performed on SKOV3 cells.Release of Caspase-12 was studied by western blot technology;and BAPTA-AM of 25,50,100 μmol/L were used to chelate Ca2+ generated by nsPEF.Then,apoptotic rate of tumor cell was tested by Hoechst 33342,and expression of Caspase-12 was studied by western blot.Experimental results indicate that the concentration of Ca2+ greatly increases with nsPEF,which induces tumor cell apoptosis through endoplasmic reticulum pathway.The mechanism of tumor cell apoptosis with nsPEF is further indicated by this research results.