| 史盼影,林温文,张保国.miR-101对宫颈癌HeLa细胞辐射敏感性的影响和机制研究[J].中华放射医学与防护杂志,2016,36(12):888-892.Shi Panying,Lin Wenwen,Zhang Baoguo.Radiosensitizing effects of miR-101 on HeLa cancer cells and underlying mechanism[J].Chin J Radiol Med Prot,2016,36(12):888-892 |
| 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 |
| 投稿时间：2016-07-11 |
| DOI：10.3760/cma.j.issn.0254-5098.2016.12.002 |
| 中文关键词: miR-101 HeLa细胞 辐射敏感性 DNA修复 |
| 英文关键词:miR-101 HeLa cells Radiosensitization DNA Repair |
| 基金项目:江苏省自然科学基金面上项目（BK20141084） |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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 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. |
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