重组腺病毒Ad－HP对肝癌细胞及肝癌肺转移模型的抑制效应  

Antitumor effects of the recombinant adenovirus Ad-HP on hepatoma carcinoma cell and pulmonary metastasis model of hepatocellular carcinoma

作　　者：齐延新[1,2,3] 王浩然[1,2] 李霄[2,3] 王卓越[1,2,3] 胡宁宁[2,3] 郭欢欢[2,3] 吴娜[2,3] 金宁一 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]军事医学科学院军事兽医研究所,吉林长春130062 [3]吉林省人兽共患病预防与控制重点实验室,吉林长春130062

出　　处：《中国兽医学报》2012年第6期823-827,832,共6页Chinese Journal of Veterinary Science

基　　金：转基因生物新品种培育重大专项基金资助项目(2009ZX08006-002B);国家自然科学基金资助项目(81101140);中国博士后基金资助项目(20100481057);吉林省重大科技攻关项目(10ZDGG007)

摘　　要：为探讨重组腺病毒Ad-HP对人肝癌细胞BEL-7402的影响及其对鼠肝癌肺转移模型的抑制作用,本研究利用MTT染色法进行了Ad-HP的体外抑瘤研究,并应用Annexin V检测、Caspase活性检测、AO/EB染色和DAPI染色等方法分析了Ad-HP的抑瘤方式。另外,本研究利用鼠肝癌细胞H22,建立了肝癌肺转移动物模型,并检测了Ad-HP对肝癌肺转移的抑制作用及其对模型动物免疫的影响。结果表明,Ad-HP能够有效抑制BEL-7402细胞的生长,且其作用具有一定时效和剂效关系趋势;Ad-HP感染导致BEL-7402细胞呈现磷脂膜外翻、细胞核皱缩、细胞膜通透性增加和Caspase酶活性增强等典型凋亡特征;应用Ad-HP治疗肿瘤模型能够有效延长模型动物平均生存期,控制转移灶形成,增强NK和CTL活性,上调Th1类细胞因子水平。总之,Ad-HT能够通过诱导细胞凋亡抑制BEL-7402肿瘤细胞增殖,并且能够有效控制肝癌肺转移灶形成,延长模型动物平均生存期。The antitumor effect of the recombinant adenovirus Ad-HP on BEL-7402 hepatoma carcinoma cells and pulmonary metastasis mouse model of hepatocellular carcinoma was evaluated by MTT assay in vitro, and the antitu- mor mechanism of Ad-HP was identified by Annexin V assay, Caspase assay, AO/EB staining, DAPI staining. The anitumor and immunoloregulation effects of Ad-HP in vivo were evaluated by pulmonary metastasis mouse model of hepatocellular carcinoma H22. The results showed that Ad-HP suppressed BEL-7402 cells successfully, and the in vitro antitumor effects of Ad-HP depended on the treat time and the treat dose. The infection of Ad-HP resulted in the exposure of phosphatidylserine,the shrink of nucleus,the permeation of membrane and the activation of caspase. Furthermore, in the animal experiment, the administration of Ad-HP resulted in the prolongation of the mean surviv- al, the limitation of metastasis, the activation of NK/CTL and upregulation of Thl type cytokines. In conclusion, Ad- HP could effectively suppress BEL 7402 cells,and prolong the mean survival of the animal model.

关 键 词：NDV HN基因 PEG-3启动子 重组腺病毒 肝癌 抗肿瘤 

分 类 号：S852.65[农业科学—基础兽医学]