表达凋亡素基因的重组禽痘病毒对人喉癌细胞Hep-2的抑制效应研究  

Anti-tumor effects on human laryngeal carcinoma Hep-2 of recombinant fowlpox virus expressing chicken anemia virus Apoptin gene

作　　者：管国芳[1] 金宁一[2] 李霄[2] 孙丽丽[3] 金春顺[1] 娄玮[1] 史平[1] 郝延茹[1] 

机构地区：[1]吉林大学白求恩医学部第二临床医院耳鼻咽喉科,长春130041 [2]军事医学科学院全军基因工程重点实验室 [3]吉林省肿瘤医院头颈外科

出　　处：《临床耳鼻咽喉头颈外科杂志》2009年第6期264-266,270,共4页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

基　　金：自然科学基金重大项目(No:39893290-4-3)

摘　　要：目的:探讨含鸡贫血病病毒(CAV)凋亡素(Apoptin)基因的重组禽痘病毒vFVApoptin诱导人喉癌细胞Hep-2凋亡的作用及其作用方式。方法:用vFVApoptin感染体外培养的人喉癌细胞Hep-2,运用MTT染色法检测重组质粒对肿瘤细胞的抑制作用;并运用流式细胞仪检测肿瘤细胞线粒体跨膜电位(ΔΨm)和细胞内活性氧(ROS)水平变化情况;通过免疫印迹检测细胞色素c(Cytoc)释放;应用底物显色法检测Caspase-3/9活性。结果:vFVApoptin感染可明显抑制Hep-2肿瘤细胞,并具有下调肿瘤细胞ΔΨm,上调其ROS水平,促进Cytoc释放和激活Caspase-3/9的功能。结论:vFVApoptin通过上调Hep-2肿瘤细胞内线粒体ROS产生,造成Cytoc的释放,激活Caspase-9,进而激活Caspase-3等下游凋亡因子,最终通过线粒体途径诱导细胞凋亡抑制Hep-2肿瘤细胞。Objective：To investigate the anti tumor effects and the mechanism of the recombinant fowlpox virus expressing Apoptin gene on human laryngeal carcinoma Hep-2. Method： Hep-2 cells cultured in vitro were in- fected with vFVApoptin. The anti tumor effects on Hep 2 cells were measured through MTT staining and, the mitochondrial trans-membrane potential （△ψm） and reactive oxygen species （ROS） were analyzed by flow cytome- try. Western blot was used to detect the release of cytochrome c （Cyto c）. Caspase-3/9 activities were measured by colorimetric assay. Result：vFVApoptin could restrain Hep-2 cells significantly and, had the function of down- regulating △ψm, up-regulating ROS, promoting Cyto c release and activating Caspase-3/9. Conclusion： Cyto c were released from mitochondria by the function of up-regulating ROS of vFVApoptin. Cyto c triggered Caspase-9 and, after the activation of Caspase 9, downstream apoptotic factors, such as caspase-3, were activated. Eventu- ally, Hep-2 cells were suppressed by mitochondrial pathway apoptosis induced by vFVApoptin.

关 键 词：禽痘病毒 凋亡素基因 人喉癌细胞Hep-2 细胞凋亡 

分 类 号：R739.65[医药卫生—肿瘤]