原发性骨性关节炎膝关节软骨细胞差异表达circRNA、miRNA筛选和生物学功能及其相互作用分析  

Screening of differentially expressed circRNA and miRNA in chondrocytes of knee joint in primary osteoarthritis and biological function and interaction analysis of them

作　　者：王莹[1] 郭雄 王民[1] 杨益民[1] 任志伟[1] 尹思[1] WANG Ying;GUO Xiong;WANG Min;YANG Yimin;REN Zhiwei;YIN Si(The First Affiliated Hospital of Xi′an Jiaotong University, Xi′an 710061, China)

机构地区：[1]西安交通大学第一附属医院,西安710061 [2]西安交通大学医学部公共卫生学院

出　　处：《山东医药》2019年第21期1-6,共6页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81602811)

摘　　要：目的筛选原发性骨性关节炎(OA)膝关节软骨细胞中差异表达的环状RNA(circRNA)及miRNA,并分析其生物学功能、相关代谢通路及相互作用。方法首先用Gene Spring software V13. 0软件筛选出5例OA及5例正常对照的膝关节软骨细胞差异表达的CircRNA,然后对差异表达的CircRNA进行基因本体论(GO)及代谢通路分析(采用KEGG通路数据库);将OA和正常对照膝软骨细胞的miRNA原始数据(下载自Array Express,获取号为:EMTAB-3514)进行对数转换,然后使用Quantile方法对数据进行标准化处理,以P <0. 05,OA和正常对照膝软骨细胞的miRNA差异倍数≥2或≤0. 5为筛选条件,用R包limma中的经验贝叶斯模型(e Bayes)筛选差异表达的miRNA;用miRror工具进行差异miRNA的靶基因预测,使用KOBAS软件对找到的靶基因进行GO及代谢通路注释富集分析。用miRanda软件对0A膝软骨细胞差异表达的miRNA进行miRNA-circRNA相互作用预测分析。结果与正常软骨细胞比较,OA膝关节软骨细胞中共筛选出1 380个表达差异的circRNA,其中215个差异circRNA表达上调,1 165个表达下调。分子功能层面,差异表达的circRNA主要影响GTP酶激活剂活性、腺苷核糖核苷酸结合、钙通道调节剂活性;细胞组成层面,差异表达的circRNA主要影响细胞外基质成分、带状胶原纤维、纤维状胶原蛋白三聚体;生物学过程层面,差异表达的circRNA主要影响细胞成分形态发生、软骨细胞发育和肢体发育。差异表达的circRNA相关代谢通路与糖原生物合成、补体和凝血级联、PDGF信号途径、胶原纤维组装和其他多聚体结构有关。与正常软骨细胞比较,OA膝关节软骨细胞中共筛选出差异表达miRNA 24个,其中上调表达及下调表达的miRNA各12个。生物学过程层面,差异表达的miRNA主要参与含核碱基的化合物生物合成过程、细胞质翻译终止、神经生长和细胞发育。细胞组成层面,主要影响细胞内部分、翻译释放�Objective To screen out the differentially expressed circRNA and miRNA in knee chondrocytes of primary osteoarthritis( OA),and to analyze their biological functions,metabolic pathways,and interactions. Methods Firstly,Gene Spring software V13. 0 was used to screen out the circRNA differential gene expression profiles in knee cartilage of 5 patients with OA and 5 normal controls,and we performed GO( Gene Ontology) and KEGG( Kyoto Encyclopedia of Genes and Genomes) pathway analysis on differentially expressed circRNA. The miRNA raw data of OA cartilage and normal control cartilage was subjected to logarithmic transformation and normalized by using the Quantile method( original data was downloaded from Array Express,accession number: E-MTAB-3514). The empirical Bayes model( e Bayes) in R-packed limma was used to analyze the differences of microRNAs between OA and normal control cartilage under the screening conditions of P < 0. 05,fold change ≥2 or ≤0. 5. The target genes of differential miRNAs were predicted by using the miRror tool. For the target gene,GO and KEGG enrichment were performed by using KOBAS software. The predictive analysis of the interaction between microRNAs and circRNAs were performed by using miRanda software. Results There were 1 380 circRNAs differentially expressed in OA cartilages and normal controls,of which 215 were up-regulated and 1 165 were down-regulated. In terms of molecular function,the differentially expressed circRNAs mainly affected GTPase activator activity,adenosine ribonucleotide binding,and calcium channel modulator activity;in terms of cell component,the differentially expressed circRNAs mainly affected the composition of extracellular matrix,ribbon collagen fibers and fibrous collagen trimer;in terms of biological processes,the differentially expressed circRNAs mainly affected cell component morphogenesis,chondrocyte development and limb development. The differentially expressed circRNA-related metabolic pathways were involved in glycogen biosynthesis,complement and coag

关 键 词：环状RNA 微小RNA 原发性骨性关节炎 基因本体功能 KEGG通路数据库 

分 类 号：R684.3[医药卫生—骨科学]