卵巢子宫内膜异位症miRNA及mRNA的表达检测及调控网络分析  

Expression of miRNA&mRNA and the regulation network in ovarian endometriosis

作　　者：沈利聪[1] 杨文青[1] 板得莹 田焱[1] Shen Licong;Yang Wenqing;Ban Deying;Tian Yan(Department of Gynecology,Xiangya Hospital,Central South University,Changsha 410008,China)

机构地区：[1]中南大学湘雅医院妇科,长沙410008

出　　处：《中国医师杂志》2020年第10期1468-1473,共6页Journal of Chinese Physician

基　　金：国家自然科学基金(81801426);湖南省自然科学基金(2018JJ3857)。

摘　　要：目的采用高通量测序检测卵巢子宫内膜异位症微小RNA(microRNA,miRNA)及mRNA的表达,分析miRNA-mRNA调控网络关系,探索卵巢子宫内膜异位症的发生机制。方法回顾性分析2017年3月至2018年3月在中南大学湘雅医院因卵巢子宫内膜异位症行手术的患者20例,取异位内膜及配对在位内膜组织,提取总RNA,分别进行miRNA测序及mRNA测序,获得差异miRNA及mRNA表达谱。运用Targetscan、miRDB数据库预测差异miRNA可能结合的mRNA,并与差异mRNA取交集,获得候选mRNA,采用Cytoscape软件进行miRNA-mRNA调控网络分析,采用DAVID数据库进行基因功能富集(GO)分析和信号通路(pathway)分析。结果与在位内膜比较,异位内膜差异表达的miRNA有369个(197个上调,172个下调),差异表达的mRNA 3 765个(1 975个上调,1 790个下调)。实时荧光定量PCR(qRT-PCR)证实miR-202-5p、miR-514a-5p在异位内膜表达高于在位内膜(P<0.05),而miR-375-3p、miR-449b-5p在异位内膜表达低于在位内膜(P<0.05),与测序结果相符合。生物信息学发现与这4个miRNA相关的候选mRNA主要富集于核内甾体激素受体结合、跨膜受体及蛋白激酶激活等生物学过程。KEGG通路富集显示其参与了多种信号通路,如TGFβ信号通路、细胞黏附分子信号通路、Wnt信号通路、Rap1信号通路。结论 miRNA及mRNA在子宫内膜异位症差异表达,二者相互作用通过多种通路参与子宫内膜异位症的发生发展。Objective To reveal the expression and regulation network of microRNA(miRNA)and mRNA in ovarian endometriosis using high-throughput sequencing,and then explore the potential pathogenesis.Methods Froni January 2017 to January 2018,twenty patients with ovarian endometriosis in Xiangya I lospital of Central South University were enrolled.Ectopic endometrium and paired eutopic endometrium were collected when surgery was conducted.After total RNA was extracted,miRNA sequencing and mRNA sequencing were performed respectively.Differential miRNA and mRNA expression profiles were analyzed.Predicted mRNA were obtained by Targetscan and niiRDB databases,and then intersected with differential mRNA to obtain candidate mRNA.miRNA-mRNA regulatory network was analyzed using Cytoscape software,whereas gene ontology(GO)and pathway function enrichment were performed by DAVID database.Results Compared with eutopic endometrium,there were 369 miRNAs(197 up-regulated,172 down-regulated)and 3765 mRN As(1975 up-regulated and 1790 down-regulated)differentially expressed in ec-topic endometrium.Real time quantitative polymerase chain reaction(qRT-PCR)confirmed that the expression of mir-202-5p and inir-514a-5p in ectopic endometrium was higher than that in eutopic endometrium(P<0.05),while the expression of mir-375-3p,mir449b-5pin ectopic endometrium was lower than that in eutopic endometrium(P<0.05).Bioinformatics found that the candidate mRNA related to these four miRNAs are mainly enriched in biological processes of nuclear steroid hormone receptor binding,transmembrane receptor and protein kinasr activity,and participate in transforming growth factor β(TGF β)sig nal pathway,adhesion junction,Writ signal pathway and Rapl signal pathway.Conclusions miRNA and mRNA are differentially expressed in endometriosis,and exact important regulatory network in the development of endometriosis by involving multiple biological processes.

关 键 词：子宫内膜异位症 微RNAS 序列分析.RNA 基因调控网络 

分 类 号：R711.71[医药卫生—妇产科学] R440[医药卫生—临床医学]