Construction of HO-8910-FasL, a Cell Line with Gene Transfer of Fas L of Ovarian Cancer

Objective To construct a HO-8910 cell line with gene transfer of FasL of ovarian cancer and to give a good model for studying ovarian cancer.Methods Human FasL cDNA was inserted into reombinant retrovirus vector plasmidpLXSN by subcolone method.By using calcium phosphate-DNA co-precipitation technique,recombinant plasmid was transfected into retrovirus package cell line PA317 and the culture supernatant containing recombinant retrovirus collected to infect the cell line HO-8910 (HO-8910-FasL).The gene transferred cells were selectively cultured with G418 and identified by polymerase chain reaction PCR, and flow cytometer(FCM) techniques.Results The recombinant PL(hFasL)SN was correctly constructed and confirmed by restriction endonuclease analysis.The presence of FasL cDNA in genome of HO-8910-FasL cell line was detected by PCR and FCM, that showed FasL has raised 40.32% gene expression in HO-8910-FasL.HO-8910-FasL cells could induce apoptosis of HL-60 cells in culture assay and apoptosis of HL-60 cells was significantly increased 25.65%.Conclusion A gene transferred HO-8910-FasL cell line,which can express FasL gene highly and have biological characteristics is successfully established.