骨髓基质干细胞神经分化微小RNA表达谱研究  

MicroRNA expression profile in bone marrow stromal cells differentiation into neuron cells

作　　者：翁伟[1] 倪文飞[2] 尹丽慧[3] 吴爱悯[2] 池永龙[2] 吴建波[3] 蒋磊[3] 

机构地区：[1]浙江省湖州市第一人民医院,313000 [2]温州医学院附属第二医院 [3]温州医学院附属第一医院

出　　处：《中华实验外科杂志》2013年第12期2671-2673,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 利用基因芯片筛选出骨髓基质干细胞（BMSCs）诱导分化前后差异表达的微小RNA（miRNA）,寻找可能的神经分化特异性的miRNA.方法 从小鼠骨髓中分离骨髓基质干细胞,采用β-巯基乙醇（β-ME）、丁羟基茴香醚（BHA）、维甲酸及碱性成纤维生长因子（bFGF）定向诱导小鼠BMSCs向神经细胞方向分化,用免疫荧光染色进行鉴定.利用基因芯片技术,筛选出分化前后差异性表达的miRNAs,采用实时荧光定量聚合酶链反应（FQ-PCR）技术,对筛选出的差异miRNA进行验证,利用靶基因预测软件对表达差异最明显的miRNA进行靶基因预测.结果 （1）骨髓基质干细胞分化后,细胞呈现典型的神经细胞形态,可见神经样轴突和胞体的锥样改变,免疫荧光染色显示神经元特异性烯醇化酶（NSE）在分化后的细胞中表达.（2）小鼠骨髓基质干细胞分化后一共有18个miRNAs表达升高,4个miRNAs表达下降.FQ-PCR验证结果总体上与芯片检测的结果一致.结论 miRNA在骨髓基质干细胞神经分化过程中大量表达,它们可能对骨髓基质干细胞的神经分化进程起重要的调控作用.Objective To detect the microRNA （miRNA） gene expression profiling in bone marrow mesenchymal stem cells （BMSCs） and BMSCs-derived neuron cells and demonstratede that specific miRNAs expression is precisely regulated during BMSCs-derived neuron cells in vitro,then find out some neuron cells differentiation-specific miRNAs,and offer a fundament to thoroughly investigate the function of miRNAs in BMSCs-derived neural differentiation.Methods Mouse BMSCs were collected from bone marrow.Using β-ME,BHA,RA and basic fibroblast growth factor （bFGF）,BMSCs were induced to differentiate into neuron cells that were identified by immunofluorescence staining.The miRNA microarray technique was used to detect miRNA expression profile in BMSCs and BMSCs-derived neuron cells,and bolting different miRNAs between BMSCs and BMSCs-derived neuron cells.Differentially expressed miRNAs were confirmed through real-time fluorescent quantitative polymerase chain reaction （FQ-PCR）.Target gene prediction software was used to predict the target gene on the most differentially expressed miRNAs.Results （1） Mter BMSCs differentiated,cells showed typical neuronal morphology：the nerve-like axons and soma cone like changes.Cells were collected for immunofluorescence staining after differentiation,and the resuhs showed that NSE was expressed in differentiated cells.（2） After differentiation of BMSCs,there were 18 up-regulated miRNAs and 4 down-regulated miRNAs.Real-time qRT-PCR results revealed that the microarray data were reliable to warrant further analysis.Conclusion MiRNAs are abundantly expressed in BMSCs differentiation process,which may play an important regulatory role in the process of BMSCs differentiation into neuron cells.

关 键 词：骨髓基质干细胞 神经分化 微小RNA 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]