The Expression of GFP Reporter Gene Drived by Different Promoters in Tumor Cells Regulated by Ionizing Radiation

Objective To study the effect of ionizing radiation on the expression of liposome mediated GFP reporter gene drived by EGR - 1 gene promoter and CMV promoter, respectively in hepatoma 7402 cells. Methods EGR -1 gene promoter and CMV promoter were used to construct pcDNA3 - CMV - GFP, pcDNAS - EGR - GFP recombinant vectors, mediated by cationic liposome LipofectAMINE to transfect the hepatoma 7402 cell line. On the basis of the optimization of transfection of liposome, transfected tumor cells were treated with different concentrations of H2O2 and different dosages of γ- ray, respectively. Fluorescent microscopy and flow cytometric( FACS) methods were used to detect the GFP expression. Results Fluorescent microscopy and FACS analysis showed that reactive oxygen intermediates and ionizing radiation could induce the GFP reporter gene expression in the pcDNA3 -EGR -GFP group,and its inducibility is in a dose -dependent manner. There was no inducible effect in the pcDNA3 - CMV - GFP control group. Conclusion EGR - 1 gene promoter had an ionizing radiation - inducible characteristic , but not CMV promoter is not so.