长链非编码RNA Gm15577参与小鼠小脑神经元增殖与分化  

Long non-coding RNA Gm15577 is involved in mouse cerebellar neurogenesis

作　　者：岳永松 张伟龙[2] 刘春英[1] 牛亚梅[1] 佟伟民[1] 

机构地区：[1]中国医学科学院基础医学研究所神经科学中心北京协和医学院基础学院病理学系,100005 [2]北京协和医学院中国医学科学院肿瘤医院分子肿瘤学国家重点实验室

出　　处：《中华病理学杂志》2015年第7期504-508,共5页Chinese Journal of Pathology

基　　金：国家自然科学基金面上项目（31471343）

摘　　要：目的鉴定新的参与奈美亨综合征（NBS）小鼠模型中与小脑发育缺陷相关的长链非编码RNA（1ncRNA），并探讨其与小脑增殖分化相关的功能。方法利用芯片技术分析野生型（野生组，Nbsl^CNS-ctr）小鼠及Nbs1神经元特异性敲除（突变组，Nbsl^CNS-ctr）小鼠小脑中RNA的表达差异；利用即时荧光定量PCR技术对芯片结果加以验证；针对目的RNA，检测其全身表达及小脑中表达模式、细胞内定位以及对母基因的表达调控模式。结果IncRNA Gm15577是由神经生长调节因子一1（Negrl）基因内部的内含子区反向转录而成，特异性表达于小脑，且在其发育过程中呈动态变化趋势，而Nbs1缺失之后整体表达水平显著降低；同时，与未分化期比较，分化后的小鼠小脑原代神经前体细胞中Gm15577及其母基因Negrl均有显著升高；Gm15577敲低之后导致Negr1以及小脑增殖分化相关因子Shh与β-catenin在RNA水平的表达显著下调；人髓母细胞瘤临床样本数据分析表明Negr1基因在正常人群与肿瘤患者之间有显著的表达差异。结论从神经特异性敲除Nbs1小鼠模型出发报道了一个新的与小鼠小脑发育相关的IncRNA Gm15577，初步结果表明Gm15577通过调控Negr1进而影响小脑神经元增殖与分化进程中的Shh与β—catenin信号通路，其基因行为异常有可能与髓母细胞瘤发病有一定的相关性。Objective To identify novel lncRNAs involved in cerebellar neurogenesis using neuronal specific Nbsl-deficient （Nbsl^CNS-ctr） mouse model. Methods Microarray analysis was performed to identify differentially expressed lncRNAs between Nbsl^CNS-ctr and Nbsl^CNS-ctr mice. Expression profiles of lncRNA Gm15577 and coding gene Negrl in mice, primary cerebellar culture and cell lines were measured using RT-qPCR. Subcellular fractionation was performed to determine the subcellular localization of Gm15577. Results Gm15577 was specifically expressed in mice cerebellum in a developmentally regulated manner, which could be abolished upon Nbsl-deficiency. Gm15577 was located in the intronic region of Negrl in a reversed orientation. Gm15577 modulated the RNA expression of Negrl, Shh and β-catenin. NEGR1 had a distinct expression pattern between normal and medulloblastoma patients. Conclusion Gm15577 may modulate cerebellar granule cell proliferation and differentiation by targeting Negrl, and their dysfunctions or abnormal expression may be related to tumorigenesis of medulloblastoma.

关 键 词：小脑 Nbs1 细胞增殖 细胞分化 长链非编码RNA 

分 类 号：R596.1[医药卫生—内科学]