HBx基因转染对Huh7细胞中基因转录的影响  

The effect of HBx protein on gene transcription in Huh7 cells

作　　者：章莉[1] 郑大利[2] 黄健 韩泽广[1] 

机构地区：[1]华东理工大学生物工程学院生物化学系,上海200237 [2]上海市疾病与健康基因组学实验室-省部共建国家重点实验室培育基地国家人类基因组南方研究中心

出　　处：《肿瘤》2008年第4期282-287,共6页Tumor

基　　金：2004年国家杰出青年基金项目(编号:30425019);2005年上海市科委重点实验室建设项目(编号:05DZ22201)

摘　　要：目的:研究HBx基因对肝癌细胞Huh7中基因表达的影响,为探讨HBx在肝癌发生发展中的分子机制提供依据。方法:以pcDNA3.1b质粒为载体,将HBx转入Huh7细胞中,同时将pcDNA3.1b作空白对照。用Western印迹法证实基因转染成功。应用Affymetrix公司的人类全基因组寡核苷酸基因芯片分析转染HBx基因前后,Huh7细胞的基因表达谱。随机选择转染前后有明显差异的6个基因(3个上调,3个下调),用荧光定量PCR技术进行验证。结果:从基因芯片结果分析中发现,与对照组相比,HBx转染后Huh7细胞中明显上调的基因有88个,下调的基因有111个,这些差异基因的功能涉及多个方面。结论:HBx基因对肝癌细胞的影响是双向的,既有转录激活的作用,同时又有转录抑制的功能。这些数据对于理解HBx基因在肝癌发生发展中的作用具有重要的参考意义。Objective： To study the effect of HBx on the gene expression profile in the hepatocellular carcinoma cell line Huh7 and to provide the molecular evidence for exploring the role of HBx in the tumorigenesis of hepatoma. Methods： HBx gene was transletted into Huh7 cells carried by pcDNA3.1 b plasmid which contained the entire open reading frame （ORF） of HBx gene. The empty vector of pcDNA3.1 b was used as a control. The HBx expression in transfected cells was confirmed by Western blotting as compared with those transfected with empty vectors. And then a microarray analysis was performed to detect the gene expression profile in Huh-7 cells by Human Genome U133 Plus 2.0 Array of Affymetrix-GeneChip. Six differentiated expressed genes （3 up-regulated and 3 downregulated） were selected randomly and verified with RFQ-PCR. Results： The microarray analysis revealed that 88 genes were significantly up-regulated and 111 genes were down-regulated in Huh7 cells after transfection with HBx gene as compared with those transfected with empty vector. The functions of these differentiated genes were related with many aspects. Conclusion： HBx affected hepatocellular carcinoma cell HUH-7 bi-directionally. It not only had transcription-activating effects but also had transcription-suppressing effects. These data had important value for understanding the role of HBx in tumorigenesis and development of hepatoma.

关 键 词：癌 肝细胞 转录激活因子类 寡核苷酸序列分析 HBX基因 

分 类 号：R730.3[医药卫生—肿瘤]