论文部分内容阅读
目的:评价RNA干扰(RNAi)同时沉默胰岛素样生长因子-1受体(IGF-1R)和表皮生长因子受体(EG-FR)后生物学特性变化及对鼻咽癌CNE2细胞株放疗敏感性的改变。方法:构建荧光标记的靶向IGF-1R及EGFR信使RNA真核表达质粒,质粒转染鼻咽癌CNE2细胞株,共聚焦显微镜观察细胞转染结果,行平板克隆实验检测细胞存活分数;流式细胞术检测细胞凋亡及细胞周期;Western-blot法检测细胞周期相关蛋白cdk4/6,cyclin D1和c-myc。结果:成功构建真核表达质粒;siRNA-IGF-1R和EGFR组的细胞存活分数显著降低(P<0.05),与对照组相比,凋亡率显著增高(P<0.05),G0/G1期细胞比例明显增多,而S期和G2/M期细胞比例明显减少,细胞周期相关蛋白cdk4/6,cyclin D1和c-myc蛋白表达降低(P<0.05)。结论:RNA同时干扰沉默IGF-1R和EGFR可以引起鼻咽癌细胞周期蛋白表达降低,细胞周期阻滞,促进细胞凋亡。
Objective: To evaluate the biological characteristics of RNA interference (RNAi) simultaneous silencing of insulin-like growth factor-1 receptor (IGF-1R) and epidermal growth factor receptor (EG-FR) and the radiosensitivity to nasopharyngeal carcinoma CNE2 cell line Change. METHODS: Fluorescently labeled eukaryotic expression plasmids targeting IGF-1R and EGFR messenger RNA were constructed. The plasmids were transfected into CNE2 cell lines of nasopharyngeal carcinoma. The transfection results were observed by confocal microscopy. Cell viability was detected by plate clone assay. Flow cytometry Cell apoptosis and cell cycle were detected by flow cytometry. The expression of cdk4 / 6, cyclin D1 and c-myc were detected by Western-blot. Results: The eukaryotic expression plasmid was constructed successfully. The cell viability of siRNA-IGF-1R and EGFR groups was significantly lower than that of the control group (P <0.05) The proportion of cells in S phase and G2 / M phase decreased significantly, and the expressions of cdk4 / 6, cyclin D1 and c-myc protein decreased (P <0.05). Conclusion: Simultaneous silencing of IGF-1R and EGFR by RNA can cause the decrease of cell cycle protein, cell cycle arrest and apoptosis in NPC cells.