Recombinant Apoptin Eukaryotic Expression Vector Inducing Apoptosis in Human Ovarian Cancer Cells

Objective To investigate effect of apoptin gene on human ovarian cancer cell(CoC1).Methods pMD18-CAVP3 and pcDNA3.1(+) were respectively double digested by restriction endonuclease KpnⅠ and XbaⅠ,and obtained 365 bp and 5.0 kb fragment,then ligatated them and the recombinant apoptin eukaryotic expression vector pcDNA3.1-VP3 was constructed.Two plasmids,pcDNA3.1(+)and pcDNA3.1-VP3,were transfected into human ovarian cancer cells(CoC1) by Lipofectamine~(TM) 2000-mediated gene transfection method in vitro.The expression of apoptin gene at transcription level was proved by RT-PCR.Apoptosis in CoC1 was observed by flow cytometry.Results The result of sequencing is consistent with reports in Genbank,apoptin gene was successfully cloned into pcDNA(3.1)(+)and recombinant apoptin eukaryotic expression vector pcDNA3.1-VP3 was successfully constructed.The transient expression of apoptin gene at transcription level had been existed in transfected cells.Apotosis rate in pcDNA3.1-VP3-transfected cells was significantly higher than that in pcDNA(3.1)(+)-transfected cells.Conclusion Apoptin induces apoptosis in human ovarian cancer CoC1.