Construction of pcDNA3/HIF-1α Expression Plasmid and its Stable Expression in Liver Cancer Cell Line HepG2

Objective To construct the plasmid of HIF-1α and transfect it into human liver cancer cell line HepG2 with liposome,to establish the cell model stably expressing HIF-1α and provide an effective tool for liver cancer research.Methods The function region of HIF-1α mRNA in HepG2 was amplified with RT-PCR and subcloned into the vector pcDNA3,the cell lines that stably expressed HIF-1α were selected with G418 pcDNA3/HIF-1α after transfected into HepG2,the expression of HIF-1α protein was detected with Western blot and immunofluorescence staining.Results The constructed expression plasmid was identified with restriction enzyme digestion and gel elecrrophoresis,two DNA bands at 5.3kb and 2.55kb were found,respectively,which was in accordance with expected results,the sequence of cloned HIF-1α gene was the same as that reported in GeneBank.Eukaryotic expression plasmid was transfected into cultured HepG2 cells with liposome,the expression of HIF-1α protein detected with Western blot and immunofluorescence staining was significantly increased.Conclusion HepG2 with stable HIF-1α expression was successfully established,which may be an effective tool for liver cancer research.