Objective To investigate the radio sensitivity of silencing HK2 gene through siRNA interference in human breast cancer MDA-MB231 cells. Methods The cells were divided into three groups: the blank group( Control) ,the negative control group ( Negative) and the test group ( HK2-siRNA) . The HK2 siRNA was transfect-ed into MDA-MB231 cells and the Western blot and qRT-PCR tests were used to detect expressions of HK2 protein and mRNA before and after silencing. Then proliferation of cells in three groups that were exposed to different doses of X-ray (0,2,4,6,8 Gy) was analyzed by CCK-8 experiment,and the apoptosis rate of cells irradiated by 4 Gy X rays was determined by flow cytometry. Results After silencing HK2 gene,the expressions of protein and mRNA of HK2 in the test group cells were inhibited effectively. After three groups of cells being exposed to different doses of X-ray, the survival rate of cells displayed a decreased trend in a dose-dependent manner. And compared with the negative control group,the cell survival rate in the test group was significantly decreased and the cell apoptosis rate in the test group was higher than that in the negative control group and blank group(P<0. 01). Conclusion Si-lencing HK2 gene through siRNA can significantly sensitize MDA-MB231 breast carcinoma cells to X radiation.%目的 观察小干扰RNA(siRNA)沉默己糖激酶-2(HK2)对乳腺癌细胞MDA-MB231放疗敏感性的影响.方法 实验分为3组:空白(Control)组、阴性对照(Negative)组和转染HK2 siRNA的实验(siRNA-HK2)组.将HK2 siRNA瞬时转染于MDA-MB231细胞,用Western blot和qRT-PCR分别检测转染前后HK2蛋白和mRNA表达;CCK-8实验观察不同剂量(0、2、4、6、8 Gy)X线照射下三组细胞增殖情况;流式细胞仪检测三组细胞在4 Gy照射下细胞凋亡率,观察siRNA干扰HK2对乳腺癌细胞MDA-MB231放疗敏感性的影响.结果 siRNA-HK2组HK2的蛋白和mRNA表达量均降低.三组细胞分别给予不同剂量的X线照射后,细胞存活率呈剂量依赖性递减的趋势,且siRNA-HK2组的存活率较Control组和Negative组明显下降(P<0.05).照射后,siRNA-HK2组细胞凋亡率明显高于Control组和Negative组(P<0.01).结论 沉默乳腺癌细胞HK2基因可增加其对放疗的敏感性.
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