细胞骨架在ns脉冲诱导肿瘤细胞凋亡中的作用  

Effect of Cytoskeleton in Tumor Cell Apoptosis Induced by Nanosecond Pulsed Electric Field

作　　者：郭飞[1] 姚陈果[1] 章锡明[1] 孙才新[1] 肖德友[2] 唐丽灵[2] 

机构地区：[1]重庆大学输配电装备及系统安全与新技术国家重点实验室,重庆400030 [2]重庆大学生物工程学院,重庆400030

出　　处：《高电压技术》2012年第1期205-210,共6页High Voltage Engineering

基　　金：国家自然科学基金(50637020;50877083);教育部博士点基金(20100191110019)~~

摘　　要：为了研究细胞骨架在ns脉冲诱导肿瘤细胞凋亡中的作用,将脉冲电场参数组合(电场强度10kV/cm,脉宽500ns,脉冲30个,频率1Hz)作用于细胞骨架裂解的Hep-G2细胞。利用Annexin-V和碘化丙碇(propidium i-odide,PI)双染法检测细胞凋亡和坏死情况;采用荧光分光光度计观测线粒体跨膜电位和细胞膜穿孔的变化情况。试验发现:肌动蛋白裂解后脉冲处理组的细胞凋亡和坏死大大降低(与单独脉冲处理组相比检验水平P<0.05),早期细胞凋亡由(16.94±2.87)%下降到(4.77±1.87)%;晚期凋亡和坏死由(20.94±3.09)%下降到(14.38±2.60)%;脉冲处理组和肌动蛋白裂解后脉冲处理组的细胞线粒体跨膜电位都出现下降,但后者的线粒体跨膜电位却明显高于前者(P<0.05);单独脉冲处理组和细胞骨架缺失后脉冲处理组二者在相同时间点时其PI荧光强度无明显差异,且PI荧光随时间而变化的规律亦相似。上述结果表明:细胞骨架的缺失抑制了ns脉冲诱导细胞凋亡的线粒体通路,进而降低细胞凋亡水平,但对细胞膜的穿孔情况无明显影响。In order to study the effect of cytoskeleton in tumor cell apoptosis induced by nanosecond pulsed electric field（nsPEF）,Parameters（field intensity of 10 kV/cm,pulse duration of 500 ns,pulse number of 30,repetition frequency of 1 Hz） of nsPEF were performed on actin-disrupted Hep-G2 cells.Tumor cell apoptosis and necrosis were studied by Annexin-V and propidium iodide（PI）,and change of the mitochondrial trans-membrane potential and cell electroporation course were also tested.Significant decreases in apoptosis and necrosis were observed in actin-disrupted nanosecond pulsed electric field（nsPEF） treated group by comparing to nsPEF treated only group（test level P0.05）,from（16.94±2.87）% to（4.77±1.87）% in early apoptosis,and（20.94±3.09）% to（14.38±2.60）% in late apoptosis and necrosis;meantime,decreases in mitochondrial trans-membrane potential were observed in both actin-disrupted nsPEF and nsPEF treated only groups,however,the latter was significantly higher than the former one（P0.05）;PI intensity at the same time and Changes in PI with time were similar in both nsPEF treated only and actin-disrupted nsPEF treated groups.The results indicated that,the loss of integrity of cytoskeleton can minimize the apoptosis induced by nsPEF through mitochondrial signaling pathway,and had no significant effect on plasma membrane electroporation.

关 键 词：ns脉冲电场(nsPEF) 肿瘤细胞 细胞骨架 Hep-G2细胞 线粒体通路 细胞凋亡 

分 类 号：R730.2[医药卫生—肿瘤]