电离辐射对不同启动子驱动的GFP报告基因表达的影响

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

目 的 研究电离辐射调控脂质体介导的EGR-1基因启动子、CMV启动子驱动的GFP报告基因在人肝癌7402细胞内的表达。 方法 利用CMV启动子、ECR-1基因启动子构建pcDNA3-CMV-GFP、pcDNA3-EGR-GFP重组质粒,经阳离子脂质体LipofectAMINE介导转染人肝癌7402细胞株,对脂质体转染条件进行优化,在最佳转染条件下,分别给予不同浓度H2O2、不同剂量γ射线处理转染细胞,用荧光显微镜、流式细胞仪(FACS)检测GFP的表达情况。 结果FACS检测显示氧自由基、电离辐射可诱导EGR-1基因启动子驱动的GFP基因表达,而不诱导CMV启动子驱动的GFP表达。 结论EGR-1基因启动子具有电离辐射诱导特性。

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.