神经细胞分化中microRNA-134靶基因网络功能分析

Analysis of microRNA-134 target gene interaction network in neural progenitor-like cell differentiation

摘要:

目的 检测microRNA-134 在神经细胞分化过程中表达量的变化，分析调控神经分化的分子机制。方法 采用表皮生长因子和成纤维细胞生长因子诱导骨髓间充质干细胞向神经细胞分化，实验分为两组：对照组不加任何因子，诱导组加入表皮生长因子和成纤维细胞生长因子。通过形态、免疫组织化学和实时荧光定量聚合酶链反应（qRT-PCR） 3 种技术鉴定骨髓间充质干细胞向神经细胞分化的情况，并通 过qRT-PCR 检测microRNA-134 在神经细胞分化中表达量的变化。利用miRWalk、DAVID、STRING和Cytoscape 生物信息学软件分别对microRNA-134 靶基因进行预测，对靶基因进行GO 功能富集分析，筛选与神经分化相关的基因进行蛋白网络功能分析。结果 MicroRNA-134 在神经细胞分化过程中表达量上升，生物信息学分析结果显示 microRNA-134 的靶基因半胱氨酸天冬氨酸蛋白酶 3（ CASP3）、整合素 β 亚基 1（ITGB1）、血小板活化因子乙酰水解酶 1b 调控子单元 1（ PAFAH1B1）、蛋白激酶 C（PRKCA）、 膜附着型本体 A（EFNA2）、NK6 同源框 1（NKX6-1）、神经细胞黏附分子（ NRCAM）、突触融合蛋白结合蛋白 1（STXBP1）、钾离子通道相互作用的蛋白2（KCNIP2）、蛋白磷酸酶ABI2、CNTN2 及FOXA1 与神经分化有关。结论 MicroRNA-134 通过作用于靶基因在神经细胞分化中起促进作用。

Abstract:

Objective To detect the expression of microRNA-134 at different time after induction and better investigate the molecular mechanism in the regulation of neural differentiation. Methods Epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) were used to induce the differentiation of bone marrow derived mesenchymal stem cells (BMSCs) into neural progenitor-like cells (NPCs). BMSCs were divided into two groups: control group (without the growth factors) and induction group (added with EGF and bFGF). The morphological changes were observed and the neuronal specific markers were detected by qRT-PCR and immunohistochemical technique. The expression of microRNA-134 was determined by qRT-PCR. The target gene interaction network was analyzed using miRWalk, DAVID, STRING and Cytoscape softwares. Results MicroRNA-134 was up-regulated after EGF/bFGF-induced differentiation of rat BMSCs, similar to the expression of microRNA-134 in the rat fetal brain. Bioinformatics analysis revealed that some target genes of microRNA-134 were associated with neuron differentiation, such as CASP3, ITGB1, PAFAH1B1, PRKCA, EFNA2, NKX6-1, NRCAM, STXBP1, KCNIP2, ABI2, CNTN2, and FOXA1. Conclusions MicroRNA-134 promotes the differentiation of BMSCs into NPCs through target genes.