山东医药Issue(7)：8-10,3.DOI:10.3969/j.issn.1002-266X.2015.07.003
稳定高表达 CYP2 E1基因肝癌HepG2细胞系的构建
Establishment of hepatocellular carcinoma cell line HepG2 stably over-expressing CYP2E1
摘要
Abstract
Objective To establish a hepatocellular carcinoma cell line HepG 2 which highly expressing CYP 2E1 stab-ly.Methods The coded sequence (CDS) of CYP2E1 was searched in NCBI website, and lentiviral vector (pLV(Exp)-Puro-CMVm-CYP2E1) was designed and constructed .293T cells were co-transfected with the vector corresponding lentivi-ral packaging plasmid ( pMDLg/pRRE, pRSV-REV and pMD2.G).Lentiviral ( Lenti-Cyp2e1-eGFP-Puro) carrying CYP2E1 gene was used to trasfect HepG2 cell line, and puromycin-resistance screening were performed to establish the HepG2 cell line that highly expressed CYP 2E1.The transfection was detected through enhanced green fluorescent protein , q-PCR and Western blotting was used to detect the expression of CYP 2E1 mRNA and protein in HepG2, no-load HepG2 cell line and HepG2 cell line transfected by CYP2E1 gene.Results HepG2 cell line all transfected CYP2E1 gene suc-cessfully.The relative expression of CYP2E1 mRNA in HepG2, no-load HepG2 cell line and HepG2 cell line transfected by CYP2E1 gene was 1.02 ±0.06, 1.06 ±0.05 and 7.42 ±0.07, the protein expression of each group was 0.26 ±0.02, 0.29 ±0.01 and 1.61 ±0.08 respectively.HepG2 cell line transfected by CYP2E1 gene had significant differences as compared with the former two groups ( all P<0.05 ) .Conclusion The hepatocellular carcinoma cell line HepG 2 stably and highly expressing CYP2E1 gene is successfully constructed by lentiviral transfection .关键词
CYP2 E1基因/慢病毒载体/肝癌/HepG2细胞Key words
CYP2E1 gene/lentiviral vector/liver carcinoma/HepG2 cells分类
医药卫生引用本文复制引用
熊永福,闫再华,杨召,李敬东..稳定高表达 CYP2 E1基因肝癌HepG2细胞系的构建[J].山东医药,2015,(7):8-10,3.基金项目
国家自然科学基金资助项目（81370531）。 （）
