| 高玲,李峰生,董波,刘丽卉,刘青杰,陈肖华,毛秉智.STAT3 RNA干扰对脑胶质瘤U251细胞活性氧含量及DNA损伤的影响[J].中华放射医学与防护杂志,2011,31(3):269-272 |
| STAT3 RNA干扰对脑胶质瘤U251细胞活性氧含量及DNA损伤的影响 |
| Effects of signal transducer and activator of transcription 3 RNAi on content of reactive oxygen species and DNA damage in glioma cell |
| 投稿时间：2010-10-29 |
| DOI：10.3760/cma.j.issn.0254-5098.2011.03.006 |
| 中文关键词: STAT3 siRNA 脑胶质瘤 活性氧 DNA损伤 |
| 英文关键词:STAT3 siRNA Glioma Reactive oxygen species DNA damage |
| 基金项目:国家自然科学基金(30770640;81001216) |
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| 中文摘要: |
| 目的 探讨STAT3 RNA干扰(RNAi)对脑胶质瘤U251细胞活性氧含量及DNA损伤的影响。方法 利用构建好的STAT3 RNAi载体(pSilencer2.1-STAT3, STAT3 组)和pSilencer2.1-GFP (RNAi对照组)分别转染U251细胞,分别检测24、48和72 h后细胞内活性氧(ROS)和丙二醛(MDA)水平;同时利用单细胞凝胶电泳方法,检测6、12和24 h后细胞内DNA损伤;利用碘化丙啶(PI)单染流式检测12、24、36和48 h后细胞周期分布情况。结果 U251细胞转染pSilencer2.1-STAT3 24 h后,细胞内ROS水平是对照的8.91倍(F =89.296, P <0.05),48 h后回复到正常水平;24、 48和72 h siSTAT3 组和siGFP组之间MDA水平无显著变化;同时,与出现明显拖尾现象的8 Gy照射U251细胞阳性对照组相比,转染pSilencer2.1-STAT3 6 h后部分细胞出现拖尾现象,12 h后拖尾变得较少,24 h后完全消失。与对照组相比,siSTAT3转染组12 h后S期的滞后率是17.22%,G2/M的滞后率是6.4%;24h后G0/G1的滞后率是18.44%;36 h后,S期的滞后率是17.99%。结论 对U251细胞STAT3 RNAi后,细胞内氧化还原状态发生改变,同时伴有DNA的损伤与修复。 |
| 英文摘要: |
| Objective To investigate the effects of s ignal transducer and activator of transcription 3 (STAT3) RNAi on the content of reactive oxygen species (ROS) and the DNA damage in glioma cells. Methods Glioma cells of the line U251 cells were cultured and transfected with STAT3 RNAi plasmid(pSilencer2.1-STAT3, STAT3 group) and pSilencer2.1-GFP (GFP control group) respectively. Part of the U251 cells were irradiated with γ-rays of 60Co as positive control group of smear phenomenon. The levels of ROS and malondialdehyde (MDA) in the cells were detected 24, 48, and 72 h later by flow cytometry and fluorescence chamoluminescence analyzer, respectively. The DNA damage in the transfected U251 cells was examined by using single cell gel electrophoresis assay, and the cell cycle distribution was examined using FACS PI staining 12, 24, and 36 h later. Results At 24 h after the transfection, the ROS level of the siSTAT3-transfected cells was 8.91 times that of the control group (F =89.296, P <0.05), and returned to the normal level 48 h later. There were not significant differences in the MDA level of the cells 24, 48, and 72 h later between the siSTAT3 group and siGFP group. Compared with the 8 Gy irradiation positive group with obvious smear phenomenon, smear phenomenon was shown in part of the cells in the siSTAT3 group 6 h later, became less 12 h later, and disappeared completely 24 h later. Compared with the control group, lag of S stage rate was 17.22% and the lag of G2/M stage rate was 6.4% 12 h later in the siSTAT-transfected group, and the G0/G1 stage lag rate was 18.44% 24 h later, and the lag of S stage rate was 17.99% 36 h later. Conclusions Inhibition of STAT3 results in the change of oxidoreduction status in glioma cells, as well as damage and reparation of DNA. |
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