人肝再生增强因子上调细胞周期素B2基因的表达  

Up-regulation effect of human augmenter of liver regeneration on transcription activity of cyclin B2 gene promoter

作　　者：邵凤娟[1] 成军[2] 马英骥[1] 郭江[2] 刘妍[3] 王巧侠[2] 

机构地区：[1]哈尔滨医科大学附属第一医院传染科,150001 [2]北京地坛医院传染病研究所 [3]解放军302医院基因研究室

出　　处：《中国地方病学杂志》2006年第6期646-649,共4页Chinese Jouranl of Endemiology

基　　金：国家自然科学基金资助项目(30371288);军队"十;五"科技攻关项目(01MB135);北京市自然科学基金项目(5042024);第35批博士后科学基金资助(2004035045)

摘　　要：目的研究人肝再生增强因子(ALR)转基因表达对细胞周期素B2启动子(cyclin B2p)转录的调节作用,从而进一步探讨ALR的生物学作用及机制。方法根据文献确定细胞周期素B2p区域,聚合酶链反应(PCR)扩增细胞周期素B2p,克隆至报告基因表达载体pCAT3-Basic中,构建pCAT3-cyclin B2p报告载体;以该质粒转染肝癌细胞系HepG2细胞系,同时与ALR的真核表达载体pcDNA3．1(-)-ALR共转染HepG2细胞系,用酶联免疫吸附法(ELISA)检测报告基因氯霉素乙酰转移酶(CAT)的表达活性。结果成功构建pCAT3- cyclin B2p报告基因表达载体;pCAT3-cyclin B2p与pcDNA3．1(-)-ALR共转染HepG2细胞系的CAT表达活性是pCAT3-Basic空载体的25．13倍,是pCAT3-cyclin B2p单转染的2．60倍。结论人ALR对细胞周期素B2p基因的转录表达具有反式激活作用。Objective To investigate the trans-regulating effect of human augmenter of liver regeneration （ALR） on transcription activity of cyclin B2 gene promoter and explore the molecular biological mechanisms of ALR. Methods Polymerase chain reaction（PCR） technique was employed to amplify the DNA sequence of cyclin B2 promoter by using HepG2 genomic DNA as template, and the product was cloned into pCAT3-Basic, the resulted vector was named as pCAT3-cyelin B2p. The HepG2 cells line were co-transfected by pCAT3-cyclin B2p and pcDNA3.1 （-）-ALR. The chloramphenicol acetyhransferase （CAT） activity in HepG2 cells transfected was detected by an enzyme linked immunosorbent assay（ELISA） kit. Results The report vector pCAT3-cyclin B2p had been constructed and confirmed by restriction enzyme digestion and sequencing. The expression of CAT in HepG2 cells eo-transfected with pCAT3-cyclin B2p and pcDNA3.1 （-）-ALR was 25.13 times as high as that of pCAT3- Basic, and 2.60 times as high as that of Hep G2 cells transfected by pCAT3-cyclin B2p. Conclusions The cyclin B2 gene promoter identified in this study has transcription activity. ALR can trorns-activate the expression of cyclin B2 gene.

关 键 词：肝再生增强因子 细胞周期素B2 启动子 反式激活 

分 类 号：R346[医药卫生—基础医学]