颈动脉粥样硬化性狭窄患者外周血miRNA-210-5P表达及其靶基因生物信息学分析  

The expression of miRNA-210-5P and bioinformatics analysis of its predicted target genes in peripheral blood of patients with carotid artherosclerotic stenosis

作　　者：黄攀 徐敏 何晓英 HUANG Pan;XU Min;HE Xiao-ying(Department of Neurology,People's Hospital of Deyang City,Deyang 618000,Sichuan,China;Department of Neurology,the Second People's Hospital of Deyang City,Deyang 618000,Sichuan,China;Department of Neurology,the Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan,China)

机构地区：[1]四川省德阳市人民医院神经内科,618000 [2]四川省德阳市第二人民医院神经内科,618000 [3]西南医科大学附属医院神经内科,泸州646000

出　　处：《中国现代神经疾病杂志》2019年第7期514-520,共7页Chinese Journal of Contemporary Neurology and Neurosurgery

基　　金：四川省应用基础研究计划项目(项目编号:14JC0166);四川省卫生厅资助项目(项目编号:110368)~~

摘　　要：目的通过检测颈动脉粥样硬化性狭窄患者外周血miRNA-210-5P相对表达量,分析miRNA-210-5P及其靶基因功能。方法选择2015年7月至2018年9月诊断明确的颈动脉粥样硬化性狭窄患者(CAS组,146例)作为观察对象,采用逆转录-聚合酶链反应(RT-PCR)检测不同处理组受试者外周血miRNA-210-5P相对表达量,并以TargetScan和CoMeTa数据库进行靶基因预测、DAVID数据库进行靶基因功能富集分析(GO分析)和KEGG信号通路分析。结果与无颈动脉粥样硬化性狭窄患者或正常受试者(对照组,60例)相比,CAS组患者血清miRNA-210-5P相对表达量升高(t=14.759,P=0.000)。其中,重度狭窄组(31例;q=23.028,P=0.000)、中度狭窄组(53例;q=6.657,P=0.000)、轻度狭窄组(62例;q=42.612,P=0.000)患者外周血miRNA-210-5P相对表达量均高于对照组;而中度狭窄组(q=34.538,P=0.000)和重度狭窄组(q=11.914,P=0.000)miRNA-210-5P相对表达量高于轻度狭窄组;重度狭窄组亦高于中度狭窄组(q=16.983,P=0.000)。ROC曲线显示,miRNA-210-5P预测颈动脉粥样硬化中至重度狭窄的曲线下面积为0.943,当最佳临界值为1.495时,其预测灵敏度90.33%、特异度92.54%;生物信息学分析提示,miRNA-210-5P潜在靶基因包括VEGFA、KCMF1、HMGCS1、KLF12、EFNA3、GIT2等54个基因;GO分析显示,miRNA-210-5P靶基因功能主要富集于血管生成、神经元发育、DNA转录因子活性的正性调控、内皮细胞趋化性、细胞迁移与分化黏附等;对KEGG信号通路的检测显示,miRNA-210-5P靶基因主要富集于突触导向信号转导通路。结论颈动脉粥样硬化性狭窄患者血清miRNA-210-5P表达上调,且可能通过调控多种靶基因而作用于突触导向信号转导通路中,参与颈动脉粥样硬化性狭窄的发生与发展。Objective To investigate the relative expression of miRNA-210-5 P in serum of patients with carotid atherosclerotic stenosis(CAS)and to explore the function of miRNA-210-5 P and its target genes using bioinformatics methods.Methods We selected 146 patients with CAS from July 2015 to September 2018 in our hospital.Reverse transcriptase-polymerase chain reaction(RT-PCR)was used to detect the relative expression of miRN A-210-5 P in peripheral blood of all enrolled patients.The target genes were predicted by using TargetScan and CoMeTa databases.The target genes of miRNA-210-5 P were enriched by Gene Ontology(GO)using DAVID data and were performed with KEGG Pathway analysis.Results Compared with non-CAS patients or normal subjects(control group,N = 60),the relative expression of miRNA-210-5 P in the serum of CAS group was significantly increased(t= 14.759,P=0.000).The relative expressions of serum miRNA-210-5 P in severe stenosis group(N=31;q=23.028,P=0.000),moderate stenosis group(N =53;q =6.657,P =0.000)and mild stenosis group(N =62;q = 42.612,P =0.000)were higher than that in control group.The relative expressions of serum miRN A-210-5 P in moderate stenosis group(q = 34.538,P = 0.000)and severe stenosis group(q = 11.914,P = 0.000)were significantly higher than that in mild stenosis group.The relative expressions of serum miRNA-210-5 P in severe stenosis group was significantly higher than that in moderate stenosis group(q= 16.983,P=0.000).Receiver operating characteristic(ROC)curve showed that the miRNA-210-5 P predicted the area under the curve(AUC)of moderates to severe stenosis in CAS to be 0.943,the sensitivity was 90.33% and the specificity was 92.54% at the best cutoff value of 1.495.Bioinformatics analysis showed there were 54 potential target genes of miRNA-210-5 P,such as VEGFA,KCMF1,HMGCS1,KLF12,EFNA3,GIT2,etc.GO analysis showed that the target genes of miRN A-210-5 P were involved in angiogenesis,neuronal development,positive regulation of DNA transcription factor activity,endothelial cell c he mo tax

关 键 词：颈动脉狭窄 动脉粥样硬化 微RNAS 基因 计算生物学 

分 类 号：R543.4[医药卫生—心血管疾病] Q811.4[医药卫生—内科学]