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目的探讨ErbB2 RNAi对脑胶质瘤U251细胞活性氧含量、DNA损伤以及细胞周期的影响。方法利用构建好的ErbB2 RNAi载体(pSilencer2.1-ErbB2)转染U251细胞后,检测不同时间点细胞内活性氧(ROS)和丙二醛(MDA)水平;同时利用单细胞凝胶电泳(SCGE)方法,检测不同时间点细胞内DNA损伤;利用PI单染检测不同时间点细胞周期分布情况。结果 U251细胞转染pSilencer2.1-ErbB2后,对细胞内ROS水平无显著性影响;U251细胞在转染psilencer2.1-ErbB2后24h时,MDA水平无显著变化,而在48h时显著性升高,72h时又恢复到正常水平;同时,U251细胞转染pSilencer2.1-ErbB2后发生DNA损伤并很快修复;ErbB2的表达抑制可引发细胞周期各时相发生紊乱。结论 U251细胞ErbB2 RNAi后,细胞内氧化还原状态发生改变,同时伴有DNA的损伤与修复以及细胞周期阻滞。
Objective To investigate the effect of ErbB2 RNAi on reactive oxygen species, DNA damage and cell cycle in glioma U251 cells. Methods U251 cells were transfected with the constructed ErbB2 RNAi vector (pSilencer2.1-ErbB2) to detect the level of reactive oxygen species (ROS) and malondialdehyde (MDA) in different time points. Single cell gel electrophoresis (SCGE) ) Method to detect DNA damage at different time points; PI single staining was used to detect cell cycle distribution at different time points. Results U251 cells transfected with pSilencer2.1-ErbB2 had no significant effect on intracellular ROS levels. U251 cells did not show significant changes in MDA levels at 24 h after transfection with psilencer2.1-ErbB2, but significantly increased at 48 h , 72h and returned to normal levels; the same time, U251 cells transfected with pSilencer2.1-ErbB2 DNA damage and rapid repair; inhibition of ErbB2 expression can lead to cell cycle disorders at various stages. Conclusion The ErbB2 RNAi of U251 cells change the redox state of cells, accompanied by DNA damage and repair and cell cycle arrest.