敲降RMI1基因加剧电离辐射诱导的DNA损伤
首发时间:2016-12-28
摘要:目的:研究RecQ解旋酶介导的基因组稳定性蛋白1(RecQ-Mediated genome instability protein 1, RMI1)在电离辐射(ionizing radiation, IR)诱导DNA损伤修复中的生物学功能。方法:运用慢病毒介导的RMI1短发夹RNA(shRNA),在细胞中特异性敲降RMI1基因,通过实时定量PCR和蛋白免疫印迹检测敲降水平,然后通过单细胞凝胶电泳技术定量分析电离辐射诱导的DNA损伤程度及修复能力的变化。结果:与对照组相比,电离辐射在敲降RMI1基因的细胞中会引起更多的DNA断裂(p<0.05)。受照15min和30min后,敲降RMI1基因的细胞中尾部DNA含量和Olive尾距均显著高于对照组细胞(p<0.05)。结论:RMI1能减少电离辐射诱导的DNA损伤程度,并且促进受损DNA的修复,在电离辐射损伤修复中具有重要的作用。
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Knocking down of RMI1 gene exacerbates DNA damage induced by ionizing radiation
Abstract:Objective To investigate the biological effect of RMI1 on DNA damage and repair induced by ionizing radiation. Methods A short hairpin RNA targeting RMI1 (shRMI1) and a negative control shRNA (shControl) were transduced into 293T cells through lentivirus vector. The levels of protein and mRNA expression of RMI1 were measured by qPCR and western blot. The level of DNA damage was tested with alkaline single cell gel electrophoresis (Comet assay). Results As compared with the shControl group, the mRNA and protein levels of RMI1 in shRMI1 cells were both decreased significantly. And Comet assay showed that the DNA damage induced by ionizing and Tail Olive moment in shRMI1 cells were significantly higher than the control group (p<0.05) at 15min and 30min after IR. Conclusion Knocking down of RMI1 enhanced the IR-induced DNA damage level in 293T cells, and decreased the efficiency of DNA repair. RMI1 may play an important role in the IR-induced DNA damage and repair.
Keywords: Ionizing radiation;RMI1 DNA damage and repair
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