Bmi-1基因对人胚胎纹状体来源神经干细胞的自我更新和增殖的作用  

Role of Bmi-1 in the self-renewal and proliferation of neural stem cells isolated from human corpus striatum

作　　者：黄爱华[1,2] 梁庆成[1] 吉慧军[3] 王旸[2] 关云谦[2] 

机构地区：[1]哈尔滨医科大学附属第二临床医学院神经内科,哈尔滨150086 [2]首都医科大学宣武医院细胞治疗中心,北京100053 [3]青岛大学附属医院神经外科,青岛266003

出　　处：《解剖学报》2010年第4期532-537,共6页Acta Anatomica Sinica

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

摘　　要：目的探讨人类纹状体来源的神经干细胞(NSCs)中,Bmi-1基因是否参与自我更新和增殖的维持。方法用human-Bmi-1干扰病毒和绿色荧光蛋白(GFP)对照病毒分别转染NSC,镜下观察转染效率。于转染后0h、48h、72h和168h4个时间点收集细胞,用实时定量聚合酶链反应检测Bmi-1干扰组与同时间点GFP对照病毒转染组细胞中的Bmi-1基因的相对转录水平,1周后比较human-Bmi-1干扰病毒和GFP对照病毒转染后细胞的克隆形成数和新生神经球直径。结果经Bmi-1干扰病毒和GFP对照病毒转染后48h,显微镜下观察见细胞均带有绿色荧光,转染效率超过90%。与转染后0h的Bmi-1基因的相对转录水平(107.3±8.0618)%相比,48h、72h和168h的Bmi-1基因的相对转录水平下降,分别为(32.7±9.74)%、(24.4±12.15)%和(32.7±10.39)%,差异有统计学意义(P<0.05)。经Bmi-1干扰后,细胞的克隆形成数和新生神经球直径都显著下降。结论 Bmi-1基因正向调控人类纹状体来源的NSC的自我更新和增殖,当细胞内的Bmi-1转录水平下降时,细胞的克隆形成数和新生神经球直径也显著下降。Objective To discuss whether Bmi-1 is participated in maintenance of the self-renewal and proliferation of neural stem cells （NSCs） isolated from fetal striatum. Methods Neural stem cells were transfected with Bmi-1 targeted shRNA-lentiviral vectors and GFP control vector respectively. At 0 hour,48 hours, 72 hours, and 168 hours after transfeetion, the Bmi-1 mRNA expression level was measured by Real-time PCR. The numbers and diameters of newly formed neurosphere were calculated under microscope. Results The transfection efficiency was shown to be more than 90% by expression of reporter gene GFP. At 0 hour,48 hours, 72 hours and 168 hours after transfection, transcription level of Bmi-1 were to be （107.3 ±8.0618）%, （32.7 ±9.74）%, （24.4 ± 12. 15）%, （32.7 ± 10.39）% compared with 0 hour, the relative expression levels of Bmi-1 at 48 hours, 72 hours, and 168 hours were significantly decreased（P 〈 0.05）. Accordingly, The number and size of newly formed neurospheres were also significantly decreased in Bmi-1 knockdown groups. Conclusion Bmi-1 may take positive roles in the maintenance of self-renewal and proliferation of NSCs isolated from human fetal striatum.

关 键 词：神经干细胞 RNA干扰 更新 增殖 实时定量聚合酶链反应 人 

分 类 号：R331.2[医药卫生—人体生理学]