单核苷酸多态rs386585341影响miR-499a-3p下游调控免疫和血管新生的网络  

Single nucleotide polymorphism rs386585341 affects the downstream network of miR-499a-3p on immunity and angiogenesis

作　　者：丁素玲 张志伟 杨茜洋 朱宝玲 张伟伟 王翔飞 杨向东 DING Suling;ZHANG Zhiwei;YANG Xiyang;ZHU Baoling;ZHANG Weiwei;WANG Xiangfei;YANG Xiangdong(Institute of Cardiovascular Diseases,Affiliated Zhongshan Hospital,Fudan University,Shanghai 200032,China;Reproductive Medicine Centre,Zhongshan Hospital,Affiliated Fudan University,Shanghai 200032,China;Fudan University Institute of Biomedical Science,Shanghai 200032,China;Department of Cardiology,Wusong Hospital,Affiliated Zhongshan Hospital,Fudan University,Shanghai 200940,China)

机构地区：[1]复旦大学附属中山医院心血管研究所 [2]复旦大学附属中山医院生殖医学中心 [3]复旦大学生物医学研究院,上海市200032 [4]复旦大学附属中山医院吴淞医院心内科,上海市200940

出　　处：《中国动脉硬化杂志》2022年第4期295-303,共9页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金项目(81970214、82170258);上海市科委基础研究项目(19JC1411400);上海市科委实验动物研究领域项目(19140902000)。

摘　　要：目的发生在microRNA基因前体或成熟序列或靶基因3′非翻译区结合位点的单核苷酸多态性(SNP)可能通过影响microRNA对靶基因的识别调控过程,参与许多疾病如肿瘤、神经系统疾病、肌肥大、心血管疾病等的发生发展过程。本研究采用生物信息学技术分析位于miR-499a-3p种子序列第4位的SNP rs386585341 A>G可能影响生物学过程及信号通路。方法应用RNAfold数据库预测rs386585341 A>G是否影响pre-miR-499a的二级结构,分别构建pre-miR-499a-A和pre-miR-499a-G过表达质粒检测rs386585341 A>G是否影响成熟miR-499a的表达,利用基因表达谱芯片和生物信息学分析rs386585341 A>G对miR-499a-3p功能的影响,取差异基因表达谱和TargetScan靶基因交集,分析rs386585341不同等位点miR-499a-3p-A和miR-499a-3p-G靶基因差异。结果rs386585341 A>G不影响pre-miR-499a的二级结构,也不影响成熟miR-499a-5p和miR-499a-3p的表达水平,但影响成熟miR-499a-3p调控的靶基因网络。取差异表达基因(DEG)谱行GO和pathway分析,发现miR-499a-3p-A和miR-499a-3p-G展示不同的生物学功能,miR-499a-3p-A的下游基因网络主要富集在免疫调控方面,而miR-499a-3p-G的下游基因网络主要富集在血管新生方面;下调表达基因和TargetScan靶基因交集分析得到4个miR-499a-3p-A直接靶基因(Spry2、Pcnx、Ndufa5及Tcf7l2),而未能得到miR-499a-3p-G直接靶基因,提示miR-499a-3p种子区域第4号位由A转换成G后,调控靶基因方式可能由促进靶基因mRNA的降解转换为仅抑制靶基因的蛋白翻译。结论rs386585341 A>G可能通过影响miR-499a-3p下游调控网络在免疫和血管新生方面发挥功能。Aim Single nucleotide polymorphisms(SNP)occurring in the precursor or mature sequences of microRNA or in their binding sites on 3′untranslated region of target genes may participate in the occurrence and development of many diseases,such as tumor,nervous system diseases,muscle hypertrophy,cardiovascular diseases and so on.In this study,bioinformatics technology was used to analyze the biological processes and signal pathways that may be affected by SNP rs386585341 A>G located at the fourth position of miR-499a-3p seed sequence.Methods RNAfold database was used to predict whether rs386585341 A>G would affect the secondary structure of pre-miR-499a.Pre-miR-499a-A and pre-miR-499a-G overexpression plasmids were constructed respectively to detect whether rs386585341 A>G would affect the expression of mature miR-499a.Gene expression microarray and bioinformatics were used to analyze the effect of rs386585341 A>G on the function of miR-499a-3p,then take the intersection of differential gene expression profile and TargetScan target gene,and analyze the divergence of target genes of miR-499a-3p with different alleles of rs386585341A>G.Results rs386585341 A>G did not affect the secondary structure of pre-miR-499a,nor the expression levels of mature miR-499a-5p and miR-499a-3p,but affected the target gene network regulated by mature miR-499a-3p.The results of GO and pathway analysis of differentially expressed genes(DEG)showed that miR-499a-3p-A and miR-499a-3p-G showed different biological functions.The downstream gene network of miR-499a-3p-A was mainly enriched in immune regulation,while the downstream gene network of miR-499a-3p-G was mainly enriched in angiogenesis.The 4 target genes of miR-499a-3p-A including Spry2,Pcnx,Ndufa5 and Tcf7l2,were obtained by intersect analysis of down-regulated genes and TargetScan target genes,but the direct target genes of miR-499a-3p-G were not obtained.It indicated that after the miR-499a-3p seed region fourth was converted from A to G by rs386585341,the target gene formula of miR-

关 键 词：单核苷酸多态性 微小RNA-499a 生物信息学 靶基因 

分 类 号：Q81[生物学—生物工程]