Construction of Eukaryotic Expression Vector for Human P115 Gene and Its Effect on Proliferation of Hepatocarcinoma Cells

Objective To construct a eukaryotic expression vector for human P115 gene and investigate the effect of the expression on proliferative activity of human hepatocarcinoma HepG2 cells.Methods Human P115 gene was amplified by RT-PCR from HepG2 cells and inserted into plasmid pEGFP-N1 Vector(+).The constructed recombinant plasmid pEGFP-N1 Vector(+)-USO1 was transfected to HepG2 cells.The transfection efficacy was determined by IFA and flow cytometry.The transfected cells were determined for expressions of P115 mRNA and protein by RT-PCR and Western blot,for proliferative activity by MTT method,and for cell cycle by flow cytometry.Results Restriction analysis and sequencing proved that recombinant plasmid pEGFP-N1 Vector(+)-USO1 was constructed correctly.The transfection efficacy of HepG2 cells with the recombinant plasmid was 84.83%.The expression levels of P115 gene and protein as well as proliferative activity of HepG2 cells transfected with the recombinant plasmid were significantly higher than those with empty vector and untransfeted.The percentage of transfected cells at G1 / G2 phases decreased significantly,while that at S phase increased significantly.Conclusion A recombinant eukaryotic expression vector for human P115 gene was successfully constructed and over-expressed in hepatocarcinoma cells,and the expressed P115 promoted the proliferation of hepatocarcinoma cells.