含人γ-干扰素基因真核表达载体的构建及其在肝癌细胞中的表达  

Expression of exogenous interferon-γ gene in human hepatocellular carcinoma cells mediated by liposome

作　　者：郭英[1] 杨丽新[1] 王坤[2] 闵军[2] 陈积圣[2] 

机构地区：[1]中山医科大学附属第三医院外科,广东广州510630 [2]孙逸仙纪念医院外科,广东广州510120

出　　处：《中国普通外科杂志》2002年第1期26-28,共3页China Journal of General Surgery

基　　金：国家自然科学基金资助 (39670 70 5)

摘　　要：目的　 构建含人γ -干扰素 (IFN -γ)基因的真核表达载体 ( pcDNA3-IFN -γ) ,并观察其在肝癌细胞株中的表达情况。方法　将IFN -γ的cDNA亚克隆于哺乳动物载体 ( pcDNA3)上 ,经脂质体介导将pcDNA3-IFN -γ导入人肝癌细胞株SMMC -772 1和QGY -770 1中 ,通过高剂量G418选择培养得出抗性克隆。RT -PCR检测IFN -γ基因在转染细胞内有无转录 ,ELISA法检测转染细胞内IFN -γ的表达水平。 结果　 IFN -γ基因的mRNA仅存在于pcDNA3-IFN -γ导入的肝癌细胞内。其细胞培养上清液存在高水平的IFN -γ。 结论　 含IFNObjective To establish a recombinant eukaryotic expression vector containing human interferon-γ(IFN-γ) cDNA,and investigate the expression of IFN-γ gene in transfected hepatocellular carcinoma (HCC) cell lines. Methods pcDNA3-IFN-γ,established by subcloning IFN-γ cDNA into a mammalian expression vector pcDNA3,was mediated into HCC cell lines SMMC-7721 and QGY-7701 by lipofectamine. The transfected HCC cells were selected in RPMI1640 containing G418(400～700μg).RT-PCR analysis and ELISA assay were used for mRNA transcription and protein expression of IFN-γ gene. Results IFN-γ mRNA was detected only in pcDNA3-IFN-γ transfected cell lines, and high levels of IFN-γ protein were detected only in the supernatants of IFN-γ gene-modified cell lines. Conclusions pcDNA3-IFN-γ can stably expressIFN-γ cDNA in the transfected HCC cell strains

关 键 词：遗传学 基因表达 重组干扰素γ 培养的肿瘤细胞 肝癌 

分 类 号：R735.702[医药卫生—肿瘤]