| 肖韡,孙新臣,秦叔逵,成红艳,张伟,曹远东,李帆.RNA干扰抑制脑胶质瘤细胞HMGA1基因表达对放射敏感性的影响[J].中华放射医学与防护杂志,2010,30(2):129-132,142 |
| RNA干扰抑制脑胶质瘤细胞HMGA1基因表达对放射敏感性的影响 |
| Suppression of brain glioma cells expression of HMGA1 by lentivirus-mediated RNAi and its effect on radiosensitivity |
| 投稿时间：2009-10-29 |
| DOI： |
| 中文关键词: RNA干扰 HMGA1 脑胶质瘤细胞 放射敏感性 |
| 英文关键词:RNA interference HMGA1 Human brain glioma Radiosensitivity |
| 基金项目:国家自然科学基金(30970792);江苏省卫生厅科研基金(H200845);江苏省135工程医学重点学科 肿瘤放射治疗学基金(K0405) |
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| 中文摘要: |
| 目的 探讨慢病毒介导的RNA干扰沉默HMGA1对脑胶质瘤细胞株SHG-44放射敏感性的影响。方法 把HMGA1siRNA慢病毒载体转染脑胶质瘤细胞株SHG-44后,用半定量RT-PCR和Western blot检测其对细胞HMGA1基因表达的影响。采用克隆形成法检测细胞对6 MV X射线的敏感性,流式细胞仪检测各组细胞凋亡率及细胞周期。结果 RT-PCR和Western blot证实,HMGA1转染组细胞存在HMGA1基因表达下调:HMGA1 mRNA的表达量转染组(0.11%±0.02%)明显低于未转染组(0.89%±0.02%, t=46.6, P＜0.01);HMGA1蛋白的表达量转染组(0.18%±0.02%)明显低于未转染组(0.86%±0.03%, t=22.6, P＜0.01)。HMGA1转染组细胞较未转染组细胞对6 MV X射线的敏感性增加,增敏比为1.73。流式细胞仪检测,转染组凋亡率(37.4%±3.1%)显著高于未转染组(6.1%±0.5%, t=12.9, P＜0.05);转染组细胞周期存在G0/G1期延迟(72.6%±2.4%)显著高于未转染组(45.2%±1.6%, t=16.2, P＜0.05))。结论 HMGA1siRNA能特异性下调细胞SHG-44中HMGA1的表达,增强了该脑胶质瘤细胞对X射线的辐射敏感性。 |
| 英文摘要: |
| Objective To investigate the radiosensitivity change of brain glioma cells of SHG-44 after HMGA1 inhibition by lentivirus-mediated RNA interference (RNAi).Methods Lentiviral vectors of HMGA1siRNA was constructed successfully and verified by PCR and DNA sequencing. After HMGA1siRNA was transfeced into the brain glioma cell line SHG-44, the expression of HMGA1 was determined by retrotranscriptase polymerase chain reaction (RT-PCR) and Western blot, respectively. The effect of radiation exposure on cells was observed by clonogenic assay. Apoptosis and cell cycle were observed by flow cytometry.Results The results of RT-PCR and Western blot indicated that the expression of HMGA1 was down-regulated obviously in HMGA1 group. The expression of HMGA1 mRNA in HMGA1 group(0.11%±0.02%) was significantly less than that in untreated group(0.89%±0.02%, t=46.6, P＜0.01). The expression of HMGA1 protein in HMGA1 group(0.18%±0.02%)was also significantly decreased compared with that in untreated group(0.86%±0.03%, t=22.6, P＜0.01). Clonogenic assay showed that HMGA1 group was more sensitive than untreated group to 6 MV X-rays 12 d post-irradiation, with SER of 1.73. Flow cytometry demonstrated that the apoptosis rate of HMGA1 group (37.4%±3.1%) was higher than that of untreated group (6.1%±0.5%, t=12.9, P＜0.05). The rate of cell cycle delay in G0/ G1 was 72.6%±2.4% in HMGA1 group and 45.2%±1.6% in untreated group with significant statistical difference(t=16.2, P＜0.05).Conclusions RNAi vectors of HMGA1 could effectively suppress the expression of HMGA1 in SHG-44 cell line, so as to enhance the radio sensitivity of brain glioma cells. |
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