Bmi-1基因对于人胚骨髓间充质干细胞增生和衰老的作用  

Bmi-1 Gene is Correlated with the Proliferation and Senescence of Mesenchymal Stem Cells Isolated from Human Fetal Bone Marrow

作　　者：王芳[1,2] 王旸[1,2] 赵春松[1,2] 关云谦[1,2] 张愚[1,2] 

机构地区：[1]首都医科大学宣武医院老年病研究所细胞治疗室 [2]教育部神经变性病重点实验室

出　　处：《首都医科大学学报》2011年第1期60-66,共7页Journal of Capital Medical University

基　　金：国家高科技研究发展计划(863计划)项目(2006AA02A114);北京市科委科技计划(D07050701350703)资助项目~~

摘　　要：目的探讨人类胚胎骨髓来源的间充质干细胞(mesenchymal stem cells,MSC)中,Bmi-1(B cell specific moloney murine leukemia virus insertion site1,Bmi-1)基因是否参与了自我更新和增生的维持。方法 用human-Bmi-1干扰病毒和GFP对照病毒分别转染MSC,进行RNA干扰(RNA interference,RNAi),镜下观察转染效率。于转染后0、72、120、168h4个时间点收集细胞,用实时定量PCR(Real-time PCR)方法检测细胞中的Bmi-1干扰组与同时间点GFP对照病毒转染组相比的相对转录水平,一周后比较human-Bmi-1干扰病毒和GFP对照病毒转染后MSC细胞的增生能力和衰老状态。结果 经human-Bmi-1干扰病毒和GFP对照病毒转染后48h,显微镜下细胞都带有绿色荧光,转染效率超过90%。与转染后0h的Bmi-1基因的相对转录水平(100±21)%相比,72、120、168h的Bmi-1基因的相对转录水平下降,分别为:(28±18)%、(55±21)%和(58±22)%,差异有统计学意义。经Bmi-1干扰后,细胞的增生能力显著下降,衰老细胞数量显著增加。结论 Bmi-1基因正性调控人类胚胎骨髓来源的MSC的增生,并防止衰老,当细胞内的Bmi-1转录水平下降时,细胞增生减慢,衰老细胞也显著增加。Objective：To understand whether Bmi-1（B cell specific moloney murine leukemia virus insertion site 1） gene participates the proliferation and senescence of mesenchymal stem cells isolated from human fetal bone marrow.Methods：Mesenchymal stem cells were transfected by lentiviral vectors encoding human-Bmi-1or GFP.At 0,72,120 and 168 h after transfection,the Bmi-1 mRNA transcription level was measured by Real-time PCR and the relative expression level was calculated by the formula：（Bmi-1/GAPDH of RNAi group）/（Bmi-1/GAPDH of GFP control）.The cell number of BrdU incorporation and senescence were observed under microscope.Results：The transfection efficiency,identified by GFP fluorescence,was more than 90% in both human-Bmi-1 RNAi group and GFP control group.At 0 h,72 h,120 h,and 168 h after transfection,the relative expression level of Bmi-1 was（100±21）%,（28±18）%,（55±21）% and（58±22）%.Compared with 0 h,the relative expression level of Bmi-1 at 72 h,120 h and 168 h was significantly decreased.Furthermore,the number of senescent cells increased significantly after Bmi-1 knockout.Conclusion：Bmi-1 participates in the maintenance of proliferation and juvenescence of MSCs,which is isolated from human fetal bone marrow.

关 键 词：间充质干细胞 BMI-1 衰老 增生 

分 类 号：R331.2[医药卫生—人体生理学]