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长链非编码 RNA KCNQ1OT1 可通过海绵吸附 miR-223-3p 在心房颤动中发挥竞争性内源性 RNA 的作用。

lncRNA KCNQ1OT1 may function as a competitive endogenous RNA in atrial fibrillation by sponging miR‑223‑3p.

机构信息

Department of Cardiology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China.

Department of Cardiology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China.

出版信息

Mol Med Rep. 2021 Dec;24(6). doi: 10.3892/mmr.2021.12510. Epub 2021 Oct 26.

Abstract

Atrial fibrillation (AF) is one of the most common forms of cardiac arrhythmia. Novel evidence has indicated that a competing endogenous RNA (ceRNA) mechanism may occur in AF. The present study aimed to identify differentially expressed microRNAs (miRNAs/miRs) in AF and predict their targeting long non‑coding RNAs (lncRNAs) to identify a potential ceRNA network involved in AF using bioinformatics analysis. The GSE68475 microarray dataset was downloaded from the Gene Expression Omnibus database and differentially expressed miRNAs in AF were obtained. In addition, right atrial appendage (RAA) tissues from patients with AF were collected to determine the expression levels of the miRNAs identified following bioinformatics analysis using reverse transcription‑quantitative PCR (n=8 per group). Subsequently, Gene Ontology (GO) functional term and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analyses of the target genes of differentially expressed miRNAs of interest were performed. The potential upstream lncRNAs targeting the identified miRNAs were predicted using bioinformatics analysis. A dual luciferase reporter assay was used to verify the existence of a targeted relationship between the differentially expressed miRNA and lncRNA of interest. The results identified 43 differentially expressed miRNAs, including 23 upregulated miRNAs. The trends in the expression levels of miR‑223‑3p were inconsistent between the microarray data and those recorded in the RAA tissues from patients with persistent AF. Therefore, miR‑223‑3p was selected as the miRNA of interest for further investigations. The target gene of miR‑233‑3p was found to be enriched in 57 GO terms and 21 KEGG signaling pathways. According to the bioinformatics prediction, 69 lncRNAs targeting miR‑223‑3p were identified, including the lncRNA growth arrest‑specific transcript 5, lncRNA KCNQ1 opposite strand/antisense transcript 1 (KCNQ1OT1) and lncRNA MYC‑induced long non‑coding RNA. The results from dual luciferase assay confirmed that miR‑223‑3p was a direct target of KCNQ1OT1. A ceRNA regulatory relationship may exist between KCNQ1OT1 and miR‑223‑3p in AF, providing therefore a novel potential research target for further studies.

摘要

心房颤动(AF)是最常见的心律失常形式之一。新的证据表明,竞争内源性 RNA(ceRNA)机制可能发生在 AF 中。本研究旨在通过生物信息学分析鉴定 AF 中差异表达的 microRNAs(miRNAs/miRs),并预测其靶向长链非编码 RNA(lncRNAs),以鉴定 AF 中涉及的潜在 ceRNA 网络。从基因表达综合数据库中下载 GSE68475 微阵列数据集,获得 AF 中的差异表达 miRNAs。此外,收集 AF 患者右心耳(RAA)组织,使用逆转录-定量 PCR (n=8 个/组)确定通过生物信息学分析鉴定的 miRNAs 的表达水平。随后,对感兴趣的差异表达 miRNAs 的靶基因进行基因本体(GO)功能术语和京都基因与基因组百科全书(KEGG)信号通路富集分析。使用生物信息学分析预测潜在的靶向鉴定 miRNAs 的上游 lncRNA。双荧光素酶报告基因检测用于验证差异表达 miRNA 和感兴趣的 lncRNA 之间存在靶向关系。结果鉴定出 43 个差异表达 miRNAs,包括 23 个上调 miRNAs。miR-223-3p 的表达水平在微阵列数据和持续性 AF 患者 RAA 组织中的趋势不一致。因此,选择 miR-223-3p 作为进一步研究的 miRNA。miR-233-3p 的靶基因富集在 57 个 GO 术语和 21 个 KEGG 信号通路中。根据生物信息学预测,鉴定出 69 个靶向 miR-223-3p 的 lncRNA,包括生长停滞特异性转录物 5、KCNQ1 反向链/反义转录物 1(KCNQ1OT1)和 MYC 诱导的长非编码 RNA。双荧光素酶报告基因检测结果证实 miR-223-3p 是 KCNQ1OT1 的直接靶标。在 AF 中,KCNQ1OT1 和 miR-223-3p 之间可能存在 ceRNA 调节关系,为进一步研究提供了一个新的潜在研究靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8db3/8569515/4bf26c2a9dd5/mmr-24-06-12510-g00.jpg

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