PC12细胞缺氧培养后DNA损伤和修复相关基因表达的研究  

A study on the expression of DNA damage and repair related genes after hypoxia in PC12 cell

作　　者：徐广润[1] 张东君[2] 张旻[1] 方思羽[1] 张苏明[1] 

机构地区：[1]华中科技大学同济医学院同济医院神经内科 [2]山东大学齐鲁医院神经内科,济南250012

出　　处：《中国康复医学杂志》2005年第3期169-171,共3页Chinese Journal of Rehabilitation Medicine

基　　金：国家自然科学基金重点项目(39730170)面上项目(39770810)

摘　　要：目的:探讨PC12细胞缺氧后细胞凋亡与DNA损伤和修复的关系,进一步阐明缺氧导致PC12细胞凋亡的机制。方法:采用TUNEL法,结合应用流式细胞术观察PC12细胞缺氧培养不同时间点细胞凋亡现象,以及DNA损伤修复相关基因表达的改变。结果:在缺氧0.5h时,开始出现凋亡细胞,此时P53、P21waf1/cip1蛋白表达也开始增高,GADD45蛋白表达达到高峰(P<0.01)。至缺氧1h凋亡细胞达高峰,此时P53、P21蛋白表达最高(P<0.05),而GADD45表达则明显下降(P<0.05)。至缺氧6—12h,则以坏死为主,此时以上的基因表达均减弱。结论:PC12细胞缺氧早期(0.5—1h),主要出现凋亡为主的细胞死亡,伴随着DNA损伤相关基因表达的动态改变,提示缺氧后PC12细胞凋亡部分是由于DNA损伤严重,损伤不能及时修复所致。Objective:To investigate the relationship between apoptosis and DNA damage after hypoxia in PC12 cell.Method:The apoptosis of PC12 cell in different time points after hypoxia was determined by TUNEL techniques,and the expression DNA damage related genes was observed by flow cytometry techniques.Result:Apoptotic cells appeared at 30min after hypoxia and the expression of protein P53,P21waf1/cip1,GADD45 began to increase.Apoptosis cell came to a head and the expression of protein P53,P21,GADD45 arrived the top at 1 hour following hypoxia(P<0.05). At 6hour and 12h after hypoxia,PC12 cells were mainly necrostic and expression of the genes also decresed.Conclusion:In the early period after hypoxia of PC12 cell(0.5-2h),cell death mostly are apoptosis companied with the increased expressions of DNA damage related genes.Therefore apoptosis of PC12cell after hypoxia partly is caused by serious DNA damage which cannot be repaired.

关 键 词：缺氧培养 修复相关基因 表达的研究 PC12细胞凋亡 相关基因表达 GADD45 DNA损伤修复 P21蛋白表达 TUNEL法 细胞凋亡现象 细胞缺氧 凋亡细胞 流式细胞术 不同时间 结合应用 细胞死亡 动态改变 P53 出现 高峰 

分 类 号：R736.6[医药卫生—肿瘤]