表达IL-24基因的溶瘤腺病毒对肝癌细胞的抑制作用  

Oncolytic adenovirus vector expressing IL-24 gene suppresses hepatocellular carcinoma in vitro

作　　者：陈漪[1] 韩丹[1] 刘彬彬[1] 梁旻[1] 孙瑞霞[1] 任正刚[1] 王艳红[1] 叶胜龙[1] 

机构地区：[1]复旦大学中山医院肝内科、复旦大学肝癌研究所,上海200032

出　　处：《中华肝胆外科杂志》2011年第3期257-260,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：基金项目：本课题受国家重点基础研究发展规划973项目资助（基金编号：2004CB518708）

摘　　要：目的研究携带IL-24基因的溶瘤腺病毒对肝癌细胞抑制生长和转移的作用。方法构建Ad．HS4．AFP．E1A／IL-24，病毒的复制由人甲胎蛋白启动子控制，携带IL-24基因。检测病毒在不同细胞系中的选择性复制，以及对高转移潜能人肝癌细胞系MHCC97-H的生长抑制、诱导凋亡和抑制转移能力（侵袭、运动、黏附）的作用。结果Ad．HS4．AFP．E1A／IL-24在肝癌细胞SMMC-7721、Hep3B和MHCC97-H中选择性表达，而不影响正常肝细胞L02（P〈0．05）。Ad．HS4．AFP．E1A／IL-24可明显抑制MHCC97-H细胞的增殖，诱导其凋亡，并抑制其体外运动、侵袭和黏附能力（P〈0．01）。RT—PCR和明胶酶谱显示其抑制肝癌作用与抑制MMP-2的表达有关。结论携带IL-24基因的溶瘤腺病毒能够选择性抑制肝癌细胞的增殖并抑制其转移。Objective To investigate the selective oncolytic role and antitumor action of a novel recombinant adenovirus containing E1A and IL-24 on hepatocellular carcinoma cell（HCC）. Methods The recombinant adenovirus expressing IL-24 （Ad. HS4. AFP. E1A/IL-24） was constructed by using modified human alpha-fetoprotein （HS4-AFP） promoter to drive adenovirus E1A gene and IL-24 gene. Cell Counting Kit-8 were performed to test the selective cytotoxicity of the virus in hepatocellular carcinoma cell lines SMMC-7721, Hep3B, MHCC97-H and hepatocyte cell line L02 . The mRNA and protein expression of IL-24 gene were detected by RT-PCR and western blot. Cell growth curves and Annexin V/PI assay were used to study cell proliferation and apoptosis of MHCC97-H. The anti-metastatic effects of the recombinant adenovirus were evaluated in cell adhesion, migration, and cell motion. Matrix metalloproteinase -2 （MMP-2） expression was examined by RT-PCR and zymography. Results Selective replications of Ad. HS4. AFP. E1A/IL-24 adenovirus were observed in over expression AFP cell line MHCC97-H, a highly metastatic potential HCC cell line but not in hepatocyte cell line 1.02. The mRNA and protein of IL-24 Were also over expressed in MHCC97-H. This recombinant adenovirus also showed the significant oncolytic action on MHCC97-H but not on L02 （P〈0.05）. Besides, the recombinant adenovirus significantly inhibited MHCC97-H metastatic potential such as cell adhesion, migration and invasion as well（P〈0.01）. Conelusion The selective oncolytic adenovirus expressing E1A and IL-24 has a selective antitumor effect and play an inhibitory role in metastasis of HCC.

关 键 词：癌 肝细胞 转移 IL-24 基因治疗 

分 类 号：R735[医药卫生—肿瘤]