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Construction and analysis of a lncRNA (PWRN2)-mediated ceRNA network reveal its potential roles in oocyte nuclear maturation of patients with PCOS.

ABSTRACT: BACKGROUND:Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women. An lncRNA, namely, Prader-Willi region nonprotein coding RNA 2 (PWRN2), was up-regulated in the cumulus cells of patients with PCOS. However, the molecular mechanism of PWRN2 in PCOS remains largely unknown. METHODS:In this study, the expression levels of PWRN2 were tested in cumulus cells through qRT-PCR analysis to confirm its potential roles in oocyte nuclear maturation of PCOS. A PWRN2-mediated ceRNA network was constructed based on three microarray datasets to investigate the molecular mechanism of PWRN2 in oocyte development of patients with PCOS. The direct interactions of the candidate genes of the ceRNA network were also demonstrated by dual-luciferase reporter assay. RESULTS:PWRN2 was found to be associated with oocyte nuclear maturation in patients with PCOS in contrast to that in normal patients. Based on the microarray data, 176 lncRNAs (118 up-regulated and 58 down-regulated) and 131 mRNAs (84 up-regulated and 47 down-regulated) were identified to be regulated by PWRN2. A PWRN2-miR-92b-3p-TMEM120B ceRNA network was constructed based on results of analysis of the combined three microarray datasets (lncRNA+mRNA microarray in KGN/shPWRN2 in this study, miRNAs microarray and lncRNA+mRNA microarray in PCOS cumulus cells reported in previous studies). The coexpression characteristics of the genes (PWRN2, miR-92b-3p and TMEM120B) were detected in the cumulus cells of cumulus-oocyte complexes at different nuclear maturity stages in PCOS. These results are in accordance with the ceRNA hypothesis. Moreover, luciferase activity assay revealed that miR-92b-3p directly binds to PWRN2 and targets TMEM120B. CONCLUSIONS:PWNR2 plays important roles in oocyte nuclear maturation in PCOS by functioning as a ceRNA to reduce the availability of miR-92b-3p for TMEM120B target binding during oocyte maturation in PCOS. Our findings would provide new information and clarify abnormal oocyte development in PCOS.

SUBMITTER: Huang X

PROVIDER: S-EPMC6091030 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature

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Construction and analysis of a lncRNA (PWRN2)-mediated ceRNA network reveal its potential roles in oocyte nuclear maturation of patients with PCOS.

Huang Xin X Pan Jiaping J Wu Bi B Teng Xiaoming X

Reproductive biology and endocrinology : RB&E 20180803 1

<h4>Background</h4>Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women. An lncRNA, namely, Prader-Willi region nonprotein coding RNA 2 (PWRN2), was up-regulated in the cumulus cells of patients with PCOS. However, the molecular mechanism of PWRN2 in PCOS remains largely unknown.<h4>Methods</h4>In this study, the expression levels of PWRN2 were tested in cumulus cells through qRT-PCR analysis to confirm its potential roles in oocyte nuclear maturation of PCOS. A ...[more]

PMID: 30075721

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Project description:Long non-coding RNAs (lncRNAs) are ncRNA transcripts >200 nucleotides that are important genetic regulators. LncRNAs can directly regulate mRNA through a lncRNA-mRNA regulatory mode and can also regulate mRNA through competitive binding to micro (mi)RNA, which is generally known as the competitive endogenous RNA (ceRNA) network. The present study evaluated the functional roles and regulatory networks of lncRNAs in chronic glomerulonephritis (CGN). The proliferative ability of mouse glomerular mesangial cells (GMCs) induced by different concentrations of lipopolysaccharide (LPS) was assessed using the Cell Counting Kit-8 assay, and RNA sequencing (RNA-seq) was performed to identify differentially expressed lncRNAs in LPS-induced GMCs. Based on the sequencing results, six lncRNAs were selected for validation using reverse transcription-quantitative PCR (RT-qPCR). Furthermore, the lncRNA-mRNA regulatory network and the lncRNA-miRNA-mRNA ceRNA network were constructed to assess the role and mechanism of CGN-related lncRNAs. To elucidate the biological functions of lncRNAs, Gene Ontology (GO) biological process term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed on all mRNAs involved in the lncRNA-mRNA regulatory network and in the ceRNA network. A total of 1,532 differentially expressed lncRNAs, including 594 upregulated lncRNAs and 938 downregulated lncRNAs, were identified using RNA-seq. The results of RT-qPCR validation were consistent with RNA-seq results. An lncRNA-mRNA regulatory network, including 236 lncRNAs and 556 mRNAs, and a ceRNA network, including 6 lncRNAs, 18 miRNAs and 419 mRNAs, were successfully constructed. The GO biological process term enrichment and KEGG pathway enrichment analyses demonstrated that those lncRNAs were often related to inflammatory response and substance metabolism. The present study identified key CGN-related lncRNAs in LPS-induced GMCs, and further demonstrated a global view of the lncRNA-mRNA regulatory network and ceRNA network involved in CGN. These results offered novel insights into the roles of lncRNAs in the pathogenesis of CGN and identified potential diagnostic biomarkers.

Dataset Information

Construction and analysis of a lncRNA (PWRN2)-mediated ceRNA network reveal its potential roles in oocyte nuclear maturation of patients with PCOS.

Publications

Construction and analysis of a lncRNA (PWRN2)-mediated ceRNA network reveal its potential roles in oocyte nuclear maturation of patients with PCOS.

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