基因表达谱芯片技术筛选乙型肝炎病毒核心蛋白结合蛋白基因C1反式调节基因  

Screening and identification of genes transactivated by C1 protein by microarray assay

作　　者：蔺淑梅[1] 成军[2] 杨媛[1] 刘敏[1] 张黎颖[2] 郭江[2] 

机构地区：[1]西安交通大学医学院第一附属医院传染科,陕西西安710061 [2]北京地坛医院传染病研究所,北京100011

出　　处：《西安交通大学学报（医学版）》2007年第2期126-129,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30571649)

摘　　要：目的对未知功能基因C1转染人肝母细胞瘤细胞系(HepG2)后的基因表达谱进行分析,探索该基因的表达对肝细胞基因表达谱的影响及其可能的调节功能线索。方法以分子生物学技术构建C1的真核表达载体pcDNA3.1(-)-C1,以表达质粒pcDNA3.1(-)-C1转染HepG2细胞,空载体pcDNA3.1(-)为平行对照,制备转染后的细胞裂解液,提取mRNA,逆转录为cDNA。应用基因表达谱芯片技术对差异表达的mRNA进行检测和分析。结果HepG2细胞经转染C1表达质粒后,有26条差异表达基因,其中24条基因表达水平下调,2条基因表达水平上调。这些差异表达的基因与细胞信号转导、凋亡、细胞增生分化及肿瘤的发生密切相关。结论应用基因表达谱芯片成功筛选了C1转染细胞后差异表达基因,为进一步阐明C1蛋白可能的生物学功能及乙型肝炎病毒核心蛋白的致病机制提供了理论依据。Objective To clarify the effect of the expression of the new gene C1 on spectra of the hepatocyte gene expression, and compare the differentially expressed genes between the HepG2 cells transfected with peDNA3.1（-）-C1 and control vector [pcDNA3. 1 （-）], respectively, by cDNA microarray technique. Methods HepG2 cells were transfected by recombined expression plasmid pcDNA3. 1 （-）-C1 and pcDNA3. 1 （-）, respectively, Total RNA was isolated from the transfected HepG2 cells with pcDNA3.1 （ - ） and pcDNA3.1 （--）-C1, respectively, cDNA was prepared by reverse transcription, cDNA microarray technique was used to detect the differentially expressed genes between the two groups. Results After screening with DNA microarray, we found that 24 genes were down-regulated while 2 genes were up-regulated, among which some genes coding proteins were involved in cell signal transduction, cell apoptosis, cell proliferation and differentiation, and tumor formation. Conclusion cDNA microarray technology is used to successfully screen the genes differentially expressed in C1- expressing HepG2 cells, which brings some new clues for studying the biological functions of C1 and infection mechanism of HBV.

关 键 词：乙型肝炎病毒 核心蛋白 反式激活基因 基因表达谱芯片 

分 类 号：R392.11[医药卫生—免疫学]