减毒水泡性口炎病毒诱导宫颈癌HeLa细胞的凋亡及其作用机制  

Attenuated vesicular stomatitis virus induces apoptosis in cervical cancer HeLa cells and its mechanism

作　　者：连海[1] 张静敏[2] 夏志平[1] 汤晶[2] 张爽[1] 于玲[1] 

机构地区：[1]军事医学科学院第十一研究所基础研究室,吉林长春130062 [2]吉林大学药学院,吉林长春130021

出　　处：《中国肿瘤生物治疗杂志》2011年第3期264-269,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.31001070);吉林省科技发展计划项目(No.20100146);军事医学科学院第十一研究所创新启动基金资助项目(No.YCX0902)~~

摘　　要：目的:研究减毒水泡性口炎病毒(vesicular stomatitis virus,VSV)对宫颈癌HeLa细胞的凋亡诱导作用,并探讨其可能的机制。方法:将减毒VSV以1.0 MOI的接种密度感染HeLa细胞,在6、12、18、24和30 h后收集细胞,MTT法检测HeLa细胞的增殖;AO/EB染色观察HeLa细胞凋亡形态学变化,Annexin V-FITC/PI双染法检测HeLa细胞早期凋亡率,流式细胞术分析HeLa细胞sub-G1凋亡峰,JC-1染色法测定细胞线粒体跨膜电位水平,caspase试剂盒检测HeLa细胞caspase-3、caspase-8及caspase-9的活性。结果:减毒VSV感染HeLa细胞12和24 h后,HeLa细胞增殖活力分别为(78.4±1.9)%和(63.1±5.6)%(P<0.01);早期凋亡细胞率分别为(16.88±2.48)%和(31.9±4.24)%(P<0.01),sub-G1凋亡峰分别为(14.85±1.48)%和(21.05±2.28)%(P<0.01)。随着减毒VSV感染时间的增加,HeLa细胞线粒体跨膜电位逐渐降低(P<0.05),caspase-9和caspase-3的活性显著升高(均P<0.05)。结论:减毒VSV能够抑制HeLa细胞的增殖,并通过caspase-9和caspase-3依赖的途径诱导HeLa细胞凋亡。Objective： To investigate the apoptosis-inducing effect of an attenuated vesicular stomatitis virus in cervical cancer HeLa cells,and to explore the possible mechanism.Methods： HeLa cells were infected with VSV（MOI=1.0） and the cell proliferation was determined by MTT assay at 6,12,18,24,and 30 h after infection.Morphological changes of apoptosis were observed by acridine orange（AO）/ethidium bromide（EB） staining.Annexin V/PI double-staining was performed to detect early apoptosis rate of HeLa cells and the sub-G1 apoptotic peak was examined by flow cytometry.The mitochondrial membrane potential of HeLa cells was measured by the JC-1 staining.The activities of caspase-9,-8 and-3 were measured by caspase assay kit.Results： After HeLa cells were exposed to attenuated VSV for 12 h and 24 h,the viabilities of cells were reduced to（78.4±1.9）% and（63.1±5.6）%（P〈0.01）;the early apoptosis rates were（16.88±2.48）% and（31.9±4.24）%（P〈0.01）;the Sub-G1 apoptotic peaks were（14.85±1.48）% and（21.05±2.28）%（P〈0.01）,respectively.Attenuated VSV significantly decreased mitochondrial membrane potential in HeLa cells with the increase of infection time（P〈0.05）.The activities of caspase-9 and caspase-3 of HeLa cells were significantly increased after VSV infection（all P〈0.05）.Conclusion： The attenuated VSV can inhibit the proliferation of HeLa cells and trigger apoptosis via caspase-9 and caspase-3-dependent pathway.

关 键 词：水泡性口炎病毒 宫颈肿瘤 HeLa细胞 凋亡 

分 类 号：R737.33[医药卫生—肿瘤] R730.54[医药卫生—临床医学]