卵巢子宫内膜异位症miRNA及mRNA的表达检测及调控网络分析

来源 :中国医师杂志 | 被引量 : 0次 | 上传用户:zhuzy0909
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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(n P<0.05),而miR-375-3p、miR-449b-5p在异位内膜表达低于在位内膜(n 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。n 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:From January 2017 to January 2018, twenty patients with ovarian endometriosis in Xiangya Hospital 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 miRDB 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 3 765 mRNAs (1 975 up-regulated and 1 790 down-regulated) differentially expressed in ectopic endometrium. Real time quantitative polymerase chain reaction (qRT-PCR) confirmed that the expression of mir-202-5p and mir-514a-5p in ectopic endometrium was higher than that in eutopic endometrium (n P<0.05), while the expression of mir-375-3p, mir-449b-5p in ectopic endometrium was lower than that in eutopic endometrium (n 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 kinase activity, and participate in transforming growth factor β (TGF β) signal pathway, adhesion junction, Wnt signal pathway and Rap1 signal pathway.n Conclusions:miRNA and mRNA are differentially expressed in endometriosis, and exact important regulatory network in the development of endometriosis by involving multiple biological processes.
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