转染RNF6基因对肝癌细胞IRS-2表达的影响  

Effect of Gene Ring Finger Protein 6 on the Expression of Insulin Receptor Substrate-2 in Hepatoma Cells

作　　者：巩健[1] 宋健[1] 

机构地区：[1]淄博职业学院制药与生物工程系,淄博255314

出　　处：《生物技术通报》2014年第6期225-228,共4页Biotechnology Bulletin

摘　　要：构建环指蛋白6(RNF6)真核表达载体,并探讨其对胰岛素受体底物1(IRS-2)表达的影响。以人cDNA为模板,PCR扩增RNF6全长编码基因,并将其克隆至载体pcDNA3.1-CHA中,将重组质粒转染肝癌细胞株HepG2,利用Real time-PCR、Western blot检测细胞内IRS-2 mRNA水平及蛋白表达情况。携带RNF6目的基因的质粒转染HepG2细胞48 h后IRS-2的mRNA表达降低,为对照组的37%,显著低于对照组,差异有统计学意义(P<0.01)。RNF6引起IRS-2的表达下调,这一过程可能由于泛素化导致胰岛素信号转导通路障碍。It was to construct an eukaryotic expression vector of ring finger protein 6（RNF6）gene and investigate the effect of RNF6 on the expression of insulin receptor substrate-2（IRS-2）. The coding sequence of hRNF6 gene was amplified by PCR with human cDNA as template. The pcDNA3.1-CHA-RNF6 was constructed and transfected into hepatocarcinoma cells（HepG2）by routine mole cular biology technology. The total RNA was extracted from HepG2 cells 72 hours post-transfection, the expression levels of IRS-2 was detected by real-time quantitative PCR. Western blotting was applied to detect the protein levels of IRS-2. Result showed that the mRNA level of IRS-2 gene in transfected HepG2 was 37%of the control. The expression level of IRS-2 was lower than the control group significantly （P〈0.01）. The expression of IRS-2 was down-regulated in HepG2 significantly, and the disorder in insulin signal transduction pathway, which may result from enhanced ubiquitylation level of IRS-2.

关 键 词：胰岛素受体底物-2 泛素化 环指蛋白6 胰岛素 

分 类 号：R735.7[医药卫生—肿瘤]