【摘要】 目的通过对调控线粒体凋亡信号通路上相关分子表达的检测,了解高强度电脉冲诱导卵巢癌组织凋亡可能的线粒体途径。方法人卵巢囊腺癌SKOV3细胞接种于裸鼠皮下建立肿瘤模型,待成瘤直径达1cm,将20只裸鼠用随机数字表法分为实验组和对照组,每组10只,能量可控脉冲治疗仪用于实验组卵巢癌移植瘤,免疫荧光法检测电压依赖阴离子通道(voltage dependent anion channel,VDAC)的变化,Fluo-3/AM探针标记后激光共聚焦显微镜定位细胞内外Ca2+分布及其含量,RT-PCR测定Bcl-2的mRNA水平。结果高强度电脉冲作用后,实验组卵巢癌细胞VDAC表达与对照组相比显著下降(P=0.025);Ca2+向线粒体流动,其含量比对照组明显增加(P=0.042);Bcl-2在高强度电脉冲作用后mRNA显著低于对照组(P=0.038)。结论高强度电脉冲作为一种强的外界刺激信号,作用于线粒体外膜VDAC,使凋亡信号的细胞传导和发生效应增强,从而导致卵巢癌组织凋亡。更多还原

【Abstract】 Objective To investigate the possible mitochondrial apoptotic signaling pathway in ovarian cancer cells induced by high intensity electric pulses.Methods Human ovarian cancer models were established in nude mice by transplanting SKOV3 cells,and then the tumors were exposed to high intensity electric pulses(through 2 electrodes,with voltage 1 kV,frequency 1 kHz,pulse width 250 ns for 1 min).The mice were sacrificed immediately after the exposure,then the mass was taken out for mitochondria extraction.The expression of cytochrome C in voltage dependant anion channel(VDAC) was detected by immuofluorescence assay.Confocal microscope were used to detect Ca2+ flowing with the aid of Fluo-3/AM staining,and RT-PCR for Bcl-2 mRNA expression.Results In experiment group,VDAC was lowered,significantly different with control group(P<0.05),Ca2+ flowed to mitochondria distinctly(P<0.05),mRNA expression of Bcl-2 fell to(0.15±0.09),much lower than the control group(P<0.05).Conclusion High intensity electric pulses can act on mitochondrial membrane,especially on VDAC,and result in the membrane potential falling and transmit intracellular signals,thus leads to the apoptosis of ovarian cancer cells.更多还原