摘要
目的研究激活的Notch1信号系统抑制体外培养的前软骨干细胞(precartilainous stem cells,PSCs)凋亡及其机制。方法用RT-PCR检测PSCs细胞中Notch信号系统的表达,并分别向培养细胞中加入Notch1激活剂rhNF-kB、抑制剂γ-secretase inhibitorⅡ(MW167)和Bcl-2抑制剂HA 14-1,空白对照加PBS缓冲液,在凋亡诱导剂Celecoxib诱导后,用MTT检测细胞活力、流式细胞术检测细胞凋亡和Western Blot检测Hes-1及Bcl-2蛋白的表达。结果PSCs细胞中存在Notch1／Jagged1信号通路,rhNF-kB组细胞活力增加,凋亡较少,Hes-1蛋白和Bcl-2蛋白表达明显增加,而MW167组、HA 14-1组和对照组细胞凋亡明显,差异有显著性意义(P<0．05),且HA 14-1组和HA 14-1协同rhNF-kB组无Bcl-2表达。结论激活的Notch1信号系统通过促进靶基因Hes-1和抗凋亡蛋白Bcl-2的表达来抑制PSCs细胞凋亡。
Objective To study the apoptosis repression in precartilaginous stem cells cultured with activated Notchl. Methods The expression of Notch was assayed by RT-PCR, then rhNF-kB, an activator of Notch, andγ-secretase inhibitor Ⅱ （MW167）, an inhibitor of Notch, and HA 14-1, an inhibitor of Bcl-2, were added into the medium of precartilaginous stem cells. PBS buffer was added to the control group. After the induction by Celecoxib, the cell viability was assessed by MTT, the apoptosis was by flowcytometry, and the proteins Hes-1 and Bcl-2 by Western Blots. Results Notch1 and Jagged1 were expressed in precartilaginous stem cells. In the group of rhNF-kB, the cell viability and the expressions of Hes-1 and Bcl-2 increased and the cell apoptosis decreased. In the MW167, HA 14-1 and control groups, the cell apoptosis increased significantly （P〈0. 05）. And there was no expression of Bcl-2 in HA 14-1 and HA 14-1 with rhNF-kB groups. Conclusions Activated Notch1 could repress the apoptosis in precartilaginous stem cells by promoting the expressions of the target gene Hes-1 and anti-apoptosis protein Bcl-2.
出处
《中华小儿外科杂志》
CSCD
北大核心
2006年第11期592-595,共4页
Chinese Journal of Pediatric Surgery
基金
国家自然科学基金资助项目(编号30371439)
