以人甲胎蛋白启动子控制表达HIV-1vpr基因的重组腺病毒诱导肝癌细胞G2期阻滞和细胞凋亡作用的研究  

Cell cycle G2 arrest and apoptosis induced by adenovirus expressing HIV-1 vpr driven by human alpha-fetoprotein promoter in hepatoma cell lines

作　　者：魏强[1] 王健伟[1] 郭丽[1] 屈建国[1] 赵玉琪[2] 洪涛[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所洪涛院士实验室,北京100052 [2]美国马里兰大学

出　　处：《中国实验诊断学》2006年第7期725-729,共5页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金(项目编号3000761748)。

摘　　要：目的研究人甲胎蛋白(AFP)启动子控制表达HIV-1 vpr基因的重组腺病毒对AFP(+)肝癌细胞G2期阻滞和细胞凋亡的作用,评估以该策略利用vpr进行肿瘤特异性基因治疗的可行性。方法将以AFP启动子控制HIV-1vpr表达的重组腺病毒rvAdAFP-vpr和空白载体rvAd-null分别感染AFP(+)的肝癌细胞BEL-7402和AFP(-)的肝癌细胞SMMC-7721,利用流式细胞仪检测Vpr的特异性表达和细胞周期分布,用细胞荧光染色和线粒体膜电位检测等方法观察细胞凋亡。结果与对照病毒rvAd-null相比,重组腺病毒rvAdAFP-vpr只在AFP(+)肝癌细胞中表达HIV-1Vpr,并诱导肝癌细胞的细胞周期G2期阻滞和细胞凋亡,而在AFP(-)肝癌细胞中不明显表达Vpr,不能显著诱导肝癌细胞G2期阻滞和细胞凋亡。结论重组腺病毒rvAdAFP-vpr对于Vpr的表达是AFP表达特异性的,可有效诱导肝癌细胞G2期阻滞和细胞凋亡,有望用于AFP(+)肝癌的基因治疗研究。Objective To investigate the cell cycle G2 arrest and apoptosis induced by adenovirus expressing HIV-1 vpr gene driven by α-fetoprotein （AFP） promoter on AFP（ ＋ ） hepatoma cell lines to assess the practibility of the strategy that utilizing HIV-1 vpr on liver cancer gene therapy.Methods The hepatoma cell lines BEL-7402, which expresses AFP [ AFP（ ＋ ） ] ,and SMMC-7721, which does not express AFP [ AFP（ - ） ], were respectively infected with the recombinant adenovims rvAdAFP-vpr that comprises HIV-1 vpr gene driven by AFP promoter, or the control adenovirus rvAd-null.The expression of Vpr and the distribution cell cycles in the hepatoma cell were examined by flow cytometery,and the apoptosis was detected by fluorescent staining as well as mitochondrial membrane potential monitoring. Results Compared to the negative control adenovirus rvAd-null, the recombinant adenovirus rvAdAFP-vpr specifically expressed HIV-1 Vpr and induced cell cycle G2 arrest and apoptosis in the AFP（ ＋ ） （BEL-7402） but not the AFP（-） （SMMC-7721）cell line.Conclusion The cell eycle G2 arrest and apoptosis in hetatoma cell lines induced by the recombinant adenovims rvAdAFP-vpr are specifically linked to the AFP expression.These encouraging results provided a helpful clue to the AFP（ ＋ ） liver cancer gene therapy.

关 键 词：甲胎蛋白启动子 HIV-1 vpr 重组腺病毒 细胞周期G2阻滞 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]