应用微阵列技术筛选HBV前-S1蛋白的反式调节基因谱  

Screening and identification of the genes trans-regulated by hepatitis B virus pre-S1 protein with microarray assay

作　　者：纪冬[1] 成军[1] 董菁[1] 刘妍[1] 王建军[1] 郭江[1] 

机构地区：[1]北京解放军第302医院,100039

出　　处：《解放军医学杂志》2004年第10期871-874,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(编号C39970674;C03011402;C30070689;C30070689);军队回国留学人员启动基金(编号98H038);全军医学科研"十五"计划青年基金项目(编号01Q138);全军医学科研"十五"计划科技攻关项目(编号01MB135)资助课题

摘　　要：目的　阐明前 S1蛋白的表达对乙型肝炎病毒 (HBV)感染肝细胞基因表达谱的影响。方法　以含有HBV全基因组的质粒G376 7(GenBank号 :AF384 371)作为模板 ,应用PCR扩增前 S1蛋白编码基因片段 ,以常规的分子生物学技术构建表达载体 pcDNA3 1(- ) preS1,以脂质体转染肝母细胞瘤细胞系HepG2 ,提取总mRNA ,逆转录为cDNA ,与转染空白表达载体pcDNA3 1(- )的HepG2细胞进行DNA芯片分析并比较。结果　在 115 2个基因表达谱的筛选中 ,发现有 30个基因表达水平显著上调 ,38个基因表达水平显著下调。结论　前 S1基因的表达对于肝细胞基因表达谱有显著影响。DNA芯片技术是分析反式调节靶基因的有效技术途径。Objective To evaluate the influence of expression of pre S1 protein on the genomic expression of hepatitis B virus (HBV) infected hepatocyte with microarray. Methods The differentially expressed genes between the hepatoblastoma cell line HepG2 transfected by pcDNA3.1(-) and pcDNA3.1(-)-preS1, were respectively compared by cDNA microarray technique. The HBV pre-S1 coding DNA fragment was amplified with polymerase chain reaction (PCR) technique by using G376-7 plasmid DNA containing the full length of HBV genome as the template. The expressive vector of pcDNA3.1-preS1 was constructed by routine molecular biological methods. HepG2 cells were transfected by pcDNA3.1(-) and pcDNA3.1-preS1, respectively, using FuGENE6 Transfection Reagent. The total RNA was isolated and reversely transcribed. Results The cDNAs were subjected for microarray screening with 1152 cDNA probes. From the scanning results, it was found that 30 genes were up-regulated and 38 genes were down-regulated by pre-S1 protein of HBV. Conclusion The expression of pre-S1 protein affected the genomic expression spectrum of HBV infected hepatocyte(HepG2 cell line).

关 键 词：HBV 基因表达谱 反式调节基因 肝细胞 蛋白编码基因 微阵列技术 PCDNA3 基因表达水平 表达载体 DNA芯片 

分 类 号：R512.62[医药卫生—内科学] R735.7[医药卫生—临床医学]