Study on E1A-TK gene transfected into human pulmonary adenocarcinoma cell A549 by lipofectamine in vitro.

Objective To transfect human pulmonary adenocarcinoma cells A549 with the eucaryotic expression plasmids pcDNA3E1A-TK by lipofectamine,and to determine the sensitivity of the transfected cells to the cytotoxic actions of ganciclovir(GCV) and X-ray.Methods Eucaryotic expression plasmids containing E1A-TK gene were transfected into human pulmonary adenocarcinoma cells A549 by lipofectamine.PCR and RT-PCR were used to examine E1A-TK gene DNA integration and mRNA expression in the transfected cells,respectively.GCV,X-ray or GCV + X-ray induced growth inhibition of the transfected cells were determined by MTT cytotoxicity assay.Results E1A-TK gene DNA integration and mRNA expression were found in the transfected cells.The IC50 of GCV and X-ray on the transfected cells was increased 113.3 and 2.9 folds,respectively.GCV and X-ray acted synergistically to increase the cell death of the transfected cells.Conclusion E1A-TK gene was successfully transfected into human pulmonary adenocarcinoma cells A549 by lipofectamine.The gene was successfully expressed and conferred sensitivity of the transfected cells to GCV and X-ray.GCV plus X-ray exhibited synergistic killing effect on the transfected cells.