miR-101对宫颈癌HeLa细胞辐射敏感性的影响和机制研究  

Radiosensitizing effects of miR-101 on HeLa cancer cells and underlying mechanism

作　　者：史盼影 林温文 张保国[1] 

机构地区：[1]苏州大学医学部放射医学与防护学院江苏省高校放射医学协同创新中心,215123

出　　处：《中华放射医学与防护杂志》2016年第12期888-892,共5页Chinese Journal of Radiological Medicine and Protection

摘　　要：Objective To study the effects of microRNA101（miR-101）on radiosensitization of human uterine cervix cancer HeLa cells and underlying mechanism. Methods HeLa cells were divided into three groups including blank control, miRNA negative control and miR-101 transfection group. The cells were irradiated by 160 kVp X-ray generated from a linear accelerator at a 目的 研究microRNA101（miR-101）对体外培养的人宫颈癌HeLa细胞辐射敏感性的影响及其作用机制.方法 实验设为3组,分别为空白对照组、阴性对照组和实验组.采用160 kVp X射线照射细胞,吸收剂量率为1.15 Gy/min.实时定量PCR（qRT-PCR）法检测miR-101的过表达情况;克隆形成实验检测miR-101对HeLa细胞辐射敏感性的影响;γ-H2AX免疫荧光法检测细胞DNA双链断裂,Western blot实验观察ATM和DNA-PKcs蛋白表达量变化.结果 转染48 h后,实验组的细胞与空白对照组相比,miR-101的表达量明显增加（t=14.16,P〈0.05）.过表达miR-101的HeLa细胞存活率明显降低（t=10.75,P〈0.05）.miR-101能增加HeLa细胞的辐射敏感性（F=7.72,P〈0.05）,辐射增敏比为1.29.γ-H2AX免疫荧光显示,miR-101能抑制辐照后细胞DNA损伤修复.过表达miR-101的HeLa细胞与对照组相比,ATM和DNA-PKcs蛋白表达量明显减少.结论 miR-101 mimic对HeLa细胞的生长有抑制作用.miR-101过表达能增加HeLa细胞的辐射敏感性,miR-101通过抑制辐照后DNA损伤修复提高辐射敏感性.dose rate of 1.15 Gy/min. Real-time quantitative PCR （qRT-PCR） was used to detect the expression of miR-101. The clonogenic survival assay was applied to evaluate the effect of miR-101 on radiosensitization of HeLa cells. γ-H2AX immunofluorescence and Western blot assays were performed to observe DNA double-strand breaks and the protein expressions of ATM and DNA-PKcs of HeLa cells, respectively. Results Compared with the negative control group, the expression of miR-101 was significantly increased in the HeLa cells at 48 h after transfection with miR-101 mimic, and the survival of HeLa cells over expression of miR-101 was significantly reduced（t=10.75,P〈0.05）. The miR-101 had remarkable radiosensitive effect on HeLa cells（F=7.72,P〈0.05） with a SERD0 of 1.29. Moreover, over-expression of miR-101 could inhibit the repair of DNA damage induced by irradiation. Compared with the control group, the protein expressions of ATM and DNA-PKcs were significantly decreased in the HeLa cells over expression of miR-101. Conclusions Over-expressions of miR-101 could inhibit cell growth and enhance radiosensitivity of HeLa cells by inhibiting the repair of radiation-induced DNA damage.

关 键 词：miR-101 HELA细胞 辐射敏感性 DNA修复 

分 类 号：R737.33[医药卫生—肿瘤] R730.55[医药卫生—临床医学]