Establishment of hepatocellular carcinoma cell line HepG2 stably over-expressing CYP2E1
XIONG Yong-fu
YAN Zai-hua
YANG Zhao
LI Jing-dong
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 .
Keywords：CYP2E1 genelentiviral vectorliver carcinomaHepG2 cells
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 8-10 )