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目的 提高基因疗法对结肠直肠肿瘤的疗效。 方法 以绿色荧光蛋白 (GFP)基因作为报告基因 ,观察在X线作用下GFP基因对结肠直肠肿瘤细胞的转染效率及表达时间 ;用克隆形成试验观察X线对胞嘧啶脱氨酶 (CD)基因转染的影响 ;同时用细胞存活率测定试验检测X线联合CD和 5 氟胞嘧啶对结肠直肠肿瘤细胞的杀伤作用。 结果 X线提高超螺旋质粒对结肠直肠肿瘤细胞的转染效率 2～ 4倍 ,可提高线性质粒转染效率 3 0倍。肿瘤细胞的存活率 :转染CD基因组LoVo细胞为 85 % ,83 4 8细胞为 88% ;剂量X线联合空载体组为 10 % ;LD90 剂量X线联合CD基因组 ,LoVo细胞为 9% ,83 4 8细胞为 11% ,加上X线作用 ,细胞总体存活率仅为 1%。 结论 X线联合自杀基因可有效地杀伤结肠直肠肿瘤细胞 ,可成为治疗结肠直肠肿瘤的一种全新有效的方法
Objective To improve the efficacy of gene therapy on colorectal tumors. Methods GFP gene was used as a reporter gene to observe the transfection efficiency and expression time of GFP gene on colorectal tumor cells under the action of X-ray. The effect of X-ray on cytosine deaminase (CD) Gene transfection. At the same time, the cytotoxicity of X-ray combined CD and 5-fluorocytosine on colorectal tumor cells was tested by cell viability assay. Results X-ray enhanced the transfection efficiency of supercoiled plasmids to colorectal tumor cells by 2-4 times, and increased the transfection efficiency of linear plasmids by 30 times. The survival rates of tumor cells were 85% in LoVo cells transfected with CD and 88% in 8348 cells, 10% in X-ray combined with empty vector group, 9% in LoVo cells with X-ray combined with LD90 4 8 cells 11%, coupled with the role of X-rays, the overall cell survival rate of only 1%. Conclusion X-ray combined with suicide gene can effectively kill colorectal tumor cells and may be a new and effective method for the treatment of colorectal tumors