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目的:研究脱氧胞苷激酶(DCK)对人宫颈癌细胞HeLa药物敏感性与辐射敏感性的影响,为宫颈癌的基因治疗提供理论依据。方法:采用FuGENE 6转染HeLa细胞构建细胞模型,实验分为空白对照组、空载体阴性对照组及DCK基因沉默(siRNA)组,实时荧光定量PCR及Western blotting检测转染前后DCK mRNA及蛋白的表达变化,细胞计数法检测细胞的增殖情况,MTT法检测细胞对阿糖胞苷(AraC)的药物敏感性,克隆形成实验检测细胞的辐射敏感性。结果:与空白对照组及阴性对照组比较,siRNA转染HeLa细胞后DCK mRNA及蛋白表达水平明显降低(P<0.05);3组细胞生长曲线无明显差异;siRNA组HeLa细胞对AraC药物的敏感性降低,辐射敏感性增高(P<0.05)。结论:抑制DCK基因表达可以降低宫颈癌细胞对药物的敏感性,增强辐射敏感性。
Objective: To study the effect of deoxycytidine kinase (DCK) on the sensitivity and radiosensitivity of human cervical cancer HeLa cells and to provide a theoretical basis for gene therapy of cervical cancer. METHODS: HeLa cells were transfected with FuGENE 6 to establish a cell model. The experiment was divided into blank control group, empty vector negative control group and DCK gene silencing (siRNA) group. Real-time fluorescent quantitative PCR and Western blotting were used to detect the expression of DCK mRNA and protein The cell proliferation was detected by the cell counting method. The cell chemosensitivity to AraC was detected by MTT assay. The radiosensitivity of the cells was detected by colony formation assay. Results: Compared with the blank control group and the negative control group, the expression of DCK mRNA and protein in HeLa cells was significantly decreased (P <0.05); the growth curve of the three groups showed no significant difference; the HeLa cells in siRNA group were sensitive to AraC Sexual decline, increased radiation sensitivity (P <0.05). Conclusion: Inhibition of DCK gene expression can reduce the sensitivity of cervical cancer cells to drugs and enhance the radiation sensitivity.