Construction of pcDNA3.0-tk eukaryotic expression vector and its induced suicidal effect on transfected human lung cancer cells

Objective To construct eukaryotic expression vector containing suicide gene HSV-tk and to investigate the cytotoxic effects of tk gene on transfected human lung cancer cells A549 after prodrug treatment.Methods HSV-tk gene fragment with EcoR Ⅰ,Xho Ⅰ restriction sites was obtained by polymerase chain reaction(PCR) and followed by cloning the fragment into the corresponding sites of pcDNA3.0 to obtain pcDNA3.0-tk eukaryotic expression vector.Recombinant plasmid pcDNA3.0-tk was transfected into A549 cells with Lipofectin-mediated method.The cytotoxic effect of a prodrug,Ganciclovir,on A549/tk cells was analyzed by cell growth curve and MTT assay.Results A549/tk cells expressed suicide tk gene on the levels of RNA and protein.Transfection with pcDNA3.0-tk conferred cytotoxicity to lung cancer cells in presence of Ganciclovir.Conclusion pcDNA3.0-tk eukaryotic expression vector has been successfully constructed,and HSV-TK/GCV system can effectively kill human lung cancer cells A549 in vitro.