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目的探讨甲基硝基亚硝基胍(MNNG)对DNA造成损伤后,对人叉头蛋白转录因子(FOXO1)核质分布的影响。方法体外培养人肺腺癌细胞系H1299,经不同浓度MNNG处理细胞后,采用彗星电泳、细胞免疫荧光、Western blot等方法,对细胞DNA损伤程度、FOXO1核质定位情况及GADD45蛋白表达变化进行检测。结果 H1299细胞DNA损伤程度随MNNG处理浓度增加而显著增强,伴随DNA损伤程度的加深,FOXO1转录因子在核内的分布逐渐增加,且在细胞DNA发生损伤4h后GADD45表达升高。结论 MNNG能造成H1299细胞DNA损伤,并促进FOXO1转录因子的核转位,从而启动损伤修复过程。
Objective To investigate the effect of methyl nitroso-nitrosoguanidine (MNNG) on DNA distribution of human forkhead protein transcription factor (FOXO1). Methods Human lung adenocarcinoma cell line H1299 was cultured in vitro. After treated with different concentrations of MNNG, the DNA damage, the localization of FOXO1 and the expression of GADD45 protein were detected by comet assay, immunofluorescence and Western blot. . Results The DNA damage degree of H1299 cells was significantly increased with the increase of MNNG concentration. With the deepening of DNA damage, the distribution of FOXO1 transcription factor increased gradually in nucleus, and the expression of GADD45 increased 4h after DNA damage. Conclusion MNNG can cause DNA damage in H1299 cells and promote the nuclear translocation of FOXO1 transcription factor, thus initiating the injury repair process.