为研究ns脉冲电场(nsPEF)治疗在体肿瘤的生物电效应机制,以接种人黑色素瘤细胞A375的BALB/c裸鼠为研究对象.采用电压幅值为4kV、脉冲宽度为200ns、重复频率为1Hz的脉冲电场进行处理。用凝胶电泳法检测脉冲处理后的DNALadder分布情况表明,与对照组相比,处理组有明显阶梯状分布,即细胞发生凋亡。用TUNEI,法检测肿瘤组织的凋亡情况表明,处理组表现出较高凋亡率(检验水准P〈0.01)。用Westernblot法和免疫组织化学方法检测脉冲处理后肿瘤组织中促凋亡蛋白Bax、凋亡抑制蛋白Bcl-2的表达量情况,2者共同表明,与对照组相比,处理组Bax表达量显著升高(P〈0.01),而Bcl-2表达量明显降低(P〈0.01)。实验结果揭示nsPEF通过Bax、Bcl-2基因调控作用来诱导肿瘤凋亡。
We experimentally investigated the bioelectric effect of nanosecond pulsed electric field (nsPEF) on in vivo tumor tissue by performing nsPEF (amplitude of 4 kV, pulse width of 200 ns, repetition frequency of 1 Hz) on BALB/c nude mouse inoculated with human melanoma cells. The distribution of DNA Ladder after the nsPEF treatment was tested by gel electrophoresis, and significant “ladder” was observed in treated group versus the eontrol group, showing the apoptosis of tumorous tissues. The apoptosis was examined by TUNEL, and an increase in apoptotic rate was observed in treated group (acceptance standard P〈0.01). Expressions of BAX and Bcl-2, which were tested by Western blot and immunochemistry method, indicate that Bax (P〈0.01) obviously increases and Bcl-2 (P〈0.01) decreases in the treated group compared with the control group. Our experimental results demonstrate that the nsPEF induced apoptosis of tumorous tissues through gene regulation affects of Bax/Bcl-2.