逆转录病毒载体介导的γ-干扰素基因在人肝癌细胞中的表达研究  

Expression of Exogenous r-Interferon Genes in Human HepatoceUuIar Carcinoma Cells Following Retroviral Transduction

作　　者：钱书兵[1] 张腾飞[1] 胡亮[1] 陈诗书[1] 

机构地区：[1]上海第二医科大学人类基因治疗研究中心上海,200025

出　　处：《中国肿瘤生物治疗杂志》1997年第2期95-99,共5页Chinese Journal of Cancer Biotherapy

基　　金：本课题由国家"863"高科技项目及国家自然科学基金项目资助

摘　　要：以逆转录病毒载体(pLXSN)将人源γ-干扰素(huIFN-γ)基因转导入4种不同的人肝癌细胞株,经G418抗性筛选均获得了阳性克隆.PCR和RT-PCR结果均表明IFN-γ基因已在基因组DNA中整合并表达.IFN-γ生物活性检测结果表明,在基因修饰的4种不同个体人肝癌细胞株及同一细胞株的5种不同克隆中,分泌的IFN-γ活性有较大差别.流式细胞仪检测细胞表面HLAⅠ类分子,结果表明基因修饰肿瘤细胞表面HLAⅠ类分子表达有显著提高.同时还首次对HLAⅠ类分子专一位点A2表达进行了分析,结果表明经IFN-γ基因修饰后,A2表达增加与HLAⅠ类分子总体增加相一致,本实验为进行基因工程修饰的肿瘤疫苗研究奠定了基础.Retroviral vector pLXSN was employed to introduce human r-Interferon (IFN-γ) gene into four human hepa-tocellular carcinoma cell lines (HCC). The G418-resistant colonies were isolated and cloned. PCR and RT-PCR analysis indicated that the integration and expression of IFN-γ gene was shown only in the transduced cells. Using a bioassay method, we found that all genetically modified HCC cells can secrete varied amount of IFN-γ. The results of flow cytome-try showed that the cell surface expression of HLA class I molecules significandy increased following transduction. Moreover , we firsdy indicated that the increase in the expression of one specific HLA class I antigen, HLA-A2, was almost in the same magnitude as that of the total HLA class I molecules after transduction with IFN-r gene.

关 键 词：逆转录病毒载体 Γ-干扰素基因 肝癌细胞 肝肿瘤 

分 类 号：R735.7[医药卫生—肿瘤]