摘要
目的:研究HBsAg基因启动子I DNA结合蛋白1(SBP1)与白介素-18(IL-18)基因表达的关系,研究SBP1在HBV致病的分子生物学机制中的作用. 方法:构建质粒pcDNA3.1(-)-SBP1,pCAT3-IL-18,转染HepG2细胞进行表达,应用酶联免疫黏附法(ELISA)检测细胞中氯霉素乙酰转移酶(CAT)的表达活性. 结果:构建的表达载体pcDNA3.1(-)-SBP1经过序列分析和酶切鉴定正确.真核表达载体pcDNA3.1(-)-SBP1和pCAT3-IL-18P共转染的HepG2细胞的pCAT表达活性是pCAT3空载体的0.37倍,是转染pCAT3-IL-18P的0.17 倍,抑制率达到86.32%. 结论:SBP1可以下调IL-18启动子的活性,抑制IL-18基因的表达.
AIM: To investigate the relationship between HBsAg promoter Ⅰ DNA binding protein 1 (SBP1) and IL-18 promoter gene expression. METHODS: Recombinant plasmid, named pcDNA3.1(-)-SBP1, were constructed by inserting a gene fragment of hepatitis B virus surface antigen promoter Ⅰ DNA binding proteinl (SBP1) into pcDNA3.1(-). Then it was transfected into HepG2 cells transfected with pCAT3-IL-18P by FuGENE 6 transfection reagents. The activity of chloramphenicol acetyltransferase (CAT) in HepG2 cells was detected by enzyme-linked immunoassay (ELISA) after 48 h. RESULTS: pcDNA3.1(-)-SBP1 was confirmed by restriction enzyme digestion and sequencing. The expression of CAT in HepG2 cells co-transfected with pCAT3-IL-18P and pcDNA3.1(-)-SBP1 was 0.37 times of that transfected with pCAT3-basic, and 0.17 times of that transfected with pCAT3-IL-18P. The inhibitory rate was about 86.32%. CONCLUSION: SBP1 can down-regulate the expression of IL-18P promoter, which provides new evidence to explain the molecular biological mechanisms of SBP1 in the interactions between IL-18P and hepatocytes.
出处
《世界华人消化杂志》
CAS
2004年第12期2797-2800,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金攻关项目
No.C03011402
No.C30070689军队九五科技攻关项目
No.98D063队回国留学人员启动基金项目
No.98H038军队十五科技攻关青年基金项目
No.01Q138军队十五科技攻关面上项目
No.01MB135~~
