白细胞介素1β转换酶基因转导人肝癌细胞株的建立及其生物学特性研究  

Biological Characteristics of the Human Hepatocellular Carcinoma Cell Line Transfected with Interleukin-lβ Converting Enzyme (ICE) Gene

作　　者：贾随旺[1] 钱其军[2] 姚晓平[2] 曹惠芳[2] 王华菁[2] 吴孟超[2] 

机构地区：[1]第一军医大学南方医院肝胆外科,广州510515 [2]第二军医大学东方肝胆外科医院,上海200433

出　　处：《中国肿瘤生物治疗杂志》1997年第4期255-258,共4页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金(39500082)资助。

摘　　要：应用基因重组技术构建了人细胞凋亡基因白介素iβ转换酶(ICE)重组逆转录病毒载体pLXSN-hICE,采用电穿孔法将其与空载体pLXSN导入包装细胞系PA317,筛选G418抗性克隆,并将其病毒上清转入人肝癌细胞株SMMC7721,经G418筛选分别获得阳性克隆SMMC7721-hICE及SMMC7721-neo.RT-PCR分析证实目的基因已整合在SMMC7721-hICE细胞基因组中.细胞生长曲线测定及裸鼠致瘤性分析表明SMMC7721-hICE体外生长速度明显低于对照细胞SMMC7721及SMMC7721-neo,而且在裸鼠体内成瘤性降低(前者3/6,后者6/6),肿瘤发生延缓,肿瘤重量明显减轻.以上结果表明,逆转录病毒介导的人ICE基因转染使肝癌细胞恶性增殖明显受抑,为开展人ICE基因治疗提供了实验基础.We used retroviral vector pLXSN to construct recombinant retroviral vectors with the human apoptosis gene, interleukin-lβ converting enzyme (ICE). The vectors were introduced into packaging cell line PA317 by electroporation method. The G418 resistant colonies were selected, and the supematants of the colonies were used to infect the human hepatocellular carcinoma cell line SMMC7721. G418 resistant colonies of SMMC7721 were named SMMC7721-MCE and SMMC7721-neo. The results of RT-PCR analysis showed that exogenous hICE gene had successfully integrated into the genome of SMMC7721-hICE cells. The proliferation rate and tumorigenicity of cells in nude mice were examined. Our data showed that the growth rate and the tumorigenicity of SMMC7721-hICE cells in nude mice were considerablely decreased comparing with parent SMMC7721 and SMMC7721-neo. These results suggested that the retroviral vector expressing hICE gene was successfully constructed and could suppress the growth ability and tumorigenicity of human hepatocellular carcinoma cells, which provided a basis for further investigation of hICE gene therapy.

关 键 词：细胞凋亡 白细胞介素1β转换酶 逆转录病毒载体 基因转染 肝癌细胞株 

分 类 号：R735.705[医药卫生—肿瘤]