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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 effects of ionizing radiation on liposome-mediated EGR-1 gene promoter and CMV promoter-driven GFP reporter gene expression in human hepatocellular carcinoma 7402 cells. Methods Recombinant plasmid pcDNA3-CMV-GFP and pcDNA3-EGR-GFP were constructed by CMV promoter and ECR-1 gene promoter. The recombinant plasmid was transfected into human hepatocellular carcinoma 7402 cell by lipofectamine. The optimal transfection conditions were given to different concentrations of H2O2, different doses of γ-ray treatment of transfected cells, fluorescence microscopy, flow cytometry (FACS) to detect the expression of GFP. Results FACS assay showed that oxygen free radicals and ionizing radiation could induce the expression of GFP gene driven by EGR-1 gene promoter without inducing GV expression driven by CMV promoter. Conclusion EGR-1 gene promoter has the characteristics of ionizing radiation.