影响骨髓间充质干细胞分化为雪旺氏细胞的转录组测序分析  

Analysis of the difference in gene expression before and after rat bone marrow mesenchymal stem cells were induced into Schwann cells

作　　者：甘泉 周子荐 王东[1] GAN Quan;ZHOU Zi⁃jian;WANG Dong(Department of Orthopedics,the Seventh Affiliated Hospital of Sun Yat⁃sen University,Shenzhen,Guangdong 518107,China)

机构地区：[1]中山大学附属第七医院骨科,广东深圳518107

出　　处：《热带医学杂志》2022年第4期464-468,604,共6页Journal of Tropical Medicine

基　　金：广东省科技计划项目国际科技合作领域(2021A0505030086)

摘　　要：目的通过转录组测序筛选影响大鼠骨髓间充质干细胞分化为雪旺氏细胞的相关差异基因。方法分离培养大鼠骨髓间充质干细胞,将所有细胞分为两组,实验组细胞体外诱导为雪旺氏细胞,对照组细胞常规培养;实验组细胞诱导完成免疫荧光鉴定S-100B与胶质纤维酸性蛋白(GFAP)表达;取两组细胞进行转录组测序,筛选出差异表达的基因,然后对其进行差异基因功能和通路显著富集性分析。结果实验组细胞诱导后GFAP及S-100B表达阳性。转录组测序结果显示,实验组与对照组细胞相比,共有3715个基因差异表达,其中表达上调基因1540个,表达下调基因2175个。基因本体(GO)分析发现差异基因富集在:(1)生物学进程:Wnt信号通路的负调控参与成骨细胞分化(P=0.000)、运动神经元迁移(P=0.000)、负调控轴突延伸参与再生(P=0.000)、跨膜受体蛋白酪氨酸磷酸酶信号通路(P=0.000)、损伤轴突生长锥形成的负调控(P=0.000)、损伤轴突侧枝萌发的正向调节(P=0.000)、神经系统发育的调节(P=0.000);(2)细胞成分:雪旺氏细胞微绒毛(P=0.000)、神经网(P=0.017);(3)分子功能S100蛋白结合(P=0.003)、神经纤毛蛋白结合(P=0.003)。KEGG Pathway分析结果发现,主要涉及细胞外基质受体相互作用通路(P=0.000)、磷肌酰肌醇-3-激酶/丝氨酸-苏氨酸蛋白激酶(PI3K-Akt)信号通路(P=0.000)、轴突导向(P=0.000)、缺氧诱导因子1(HIF-1)信号通路(P=0.000)、Wnt信号通路(P=0.000)、Ras信号通路(P=0.003)、丝裂原激活蛋白激酶(MAPK)信号通路(P=0.005)。结论骨髓间充质干细胞诱导为雪旺氏细胞过程可能主要与调节PI3KAkt、Wnt、MAPK等信号通路及轴突形成有关。Objective To screen differential genes that influence the differentiation of rat bone marrow mesenchymal stem cells into Schwann cells by transcriptome sequencing.Methods Rat bone marrow mesenchymal stem cells were isolated and cultured,and all cells were divided into two groups.The experimental group cells were induced into Schwann cells in vitro,and the control group cells were cultured conventionally.After induction,the expression of S-100B and glial fibrillary acidic protein(GFAP)was detected by immunofluorescence in the experimental group.The differentially expressed genes in these two groups were detected by transcriptome sequencing,and then the function of differentially expressed genes and the significant enrichment of pathways were analyzed.Results GFAP and S-100B expression were positive after cell induction in the experimental group.Transcriptome sequencing results showed that there were 3715 differentially expressed genes in the experimental group compared with the control group,including 1540 up-regulated genes and 2175 down-regulated genes;gene ontology(GO)analysis found that the differential genes were enriched in:(1)biological process:negative regulation of canonical Wnt signaling pathway involved in osteoblast differentiation(P=0.000),motor neuron migration(P=0.000),negative regulation of axon extension involved in regeneration(P=0.000),transmembrane receptor protein tyrosine phosphatase signaling pathway(P=0.000),negative regulation of formation of growth cone in injured axon and regulation of nervous system development(P=0.000).(2)cellular component:Schwann cell microvillus(P=0.000),perineuronal net(P=0.000).(3)molecular function:S100 protein binding and neuropilin binding(P=0.003).KEGG pathway analysis showed that differential genes were mainly enriched in extracellular matrix-receptor interaction(P=0.000),phosphatidylinositol 3-kinase/serine threonine protein kinase(PI3K-Akt)signaling pathway(P=0.000),Axon guidance(P=0.000),hypoxia inducible factor-1(HIF-1)signaling pathway(P=0.000),Wnt signa

关 键 词：骨髓间充质干细胞 体外分化 雪旺氏细胞 转录组测序 基因表达 

分 类 号：R745[医药卫生—神经病学与精神病学]