NF—κB在永生化神经前体细胞凋亡中的作用  

Role of NF-κB in apoptosis of immortalized neural progenitor cells

作　　者：刘志恒[1] 桂伶俐[1] 祝畅[1] 姚文龙[1] 张传汉[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院麻醉科,武汉市430030

出　　处：《中华麻醉学杂志》2010年第1期105-108,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（30300328）

摘　　要：目的探讨NF-κB在永生化神经前体细胞凋亡中的作用。方法永生化神经前体细胞接种于6孔板，随机分为5组，每组6孔，INPC组不进行基因转染，INPC／CMV组转染表达质粒RcCMV；INPC／050组、INPC／p65组分别转染含050基因和p65基因的表达质粒RcCMV，转染后2d，加入200μg／ml G418筛选12～14d，进行阳性克隆扩大培养3—4周，检测050mRNA、p65mRNA表达、NF-κB活性和细胞凋亡情况。INPC／p50#5组转染含p65基因的表达质粒RcCMV，加入200μg／mlG418筛选12—14d，进行阳性克隆扩大培养3～4周，转染含pSO基因的表达质粒RcCMV，2d后进行上述指标的检测。结果INPC／050组和INPC／p50#5组p50mRNA表达高于其他组（P〈0．05），INPC／p65组和INPC／p50p65组p65mRNA表达、NF-κB活性及细胞凋亡率高于其他组（P〈0．05）。结论NF-κB活性升高可促讲永牛化神绎前体细胞的凋产．Objective To investigate the role of NF-κB in apoptosis of immortalized neural progenitor cells （ INPCs）. Methods INPCs were cultured in 6-well plates and were randomly divided into 5 groups （ n = 6 each） ： group I was not transfected with any plasmid （group INPC）; group II was transfected with control plasmid （group INPC/CMV） ; group m was transfeeted with plasmid RcCMV-p50 （ group INPC/p50） ; group IV was transfected with plasmid RcCMV-p65 （group INPC/p65） and groupV was transfected with plasmid RcCMV-p50 and RcCMV-p65 （group INPC/p50p65）. Group INPC/CMV （II）, INPC/p50 （m） and INPC/p65 （IV） were screened by G418, and the positive clones were then cultured for 3-4 weeks. The transcription of p50 mRNA or p65 mRNA was detected by RT-PCR. The NF--κB activity was measured by luciferase reporter gene assay. The cell apoptosis was measured by mmexin V/PI staining. In group INPC/p50p65 and group INPC/p65, the cultured positive clone was transiently transfected with plasmid RcCMV-p50. Two days after transfection, the same measurement was performed in group INPC/p50p65 and the other groups. Results The expression of p50 mRNA was significantly increased in group INPC/p50 and INPC/p50p65 as compared with the other groups （P 〈 0.05）. The expression of p65 mRNA, the NF-κB activity and the apoptotic rate were significantly increased in group INPC/p65 and INPC/p50p65 as compared with the other groups （ P 〈 0.05）. Conclusion Enhanced NF-κB activity can increase immortalized neural progenitor cell apoptosis.

关 键 词：NF-ΚB 干细胞 细胞凋亡 

分 类 号：R741[医药卫生—神经病学与精神病学]