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急性川崎病差异表达 microRNAs 的鉴定。

Identification of differentially expressed microRNAs in acute Kawasaki disease.

机构信息

Cardiology Department, Children's Hospital of Soochow University, Suzhou, Jiangsu 215025, P.R. China.

Radiology Department, Fudan University Affiliated Huashan Hospital, Shanghai 200040, P.R. China.

出版信息

Mol Med Rep. 2018 Jan;17(1):932-938. doi: 10.3892/mmr.2017.8016. Epub 2017 Nov 8.

Abstract

The present study used microarray analysis to screen the plasma expression of microRNAs (miRNAs) in patients with acute Kawasaki disease (KD) and aimed to explore the pathogenesis of KD. Plasma was collected from children with acute KD (n=6) and from healthy control children (n=6). Total RNA was extracted and differential miRNA expression between the two groups was determined. Differentially expressed miRNAs were validated using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) in an independent cohort (n=8). Target genes of the differentially expressed miRNAs were predicted and analyzed for gene ontology term enrichment and Kyoto Encyclopedia of Genes and Genomes pathways. miRNA microarray analysis revealed that seven miRNAs (miRs) were significantly upregulated (hsa‑let‑7b‑5p, hsa‑miR‑223‑3p, hsa‑miR‑4485, hsa‑miR‑4644, hsa‑miR‑4800‑5p, hsa‑miR‑6510‑5p and hsa‑miR‑765) and three were significantly downregulated (hsa‑miR‑33b‑3p, hsa‑miR‑4443 and hsa‑miR‑4515) in acute KD compared with the healthy controls. hsa‑miR‑223‑3p expression levels detected by RT‑qPCR were consistent with the microarray results. A total of 62 target genes of hsa‑miR‑223‑3p were predicted. In total, 10 differentially expressed miRNAs were identified in acute KD, of which hsa‑miR‑223‑3p was verified by RT‑qPCR.

摘要

本研究采用微阵列分析筛选急性川崎病(KD)患者血浆中 microRNAs(miRNAs)的表达,旨在探讨 KD 的发病机制。收集急性 KD 患儿(n=6)和健康对照儿童(n=6)的血浆。提取总 RNA,比较两组间差异表达的 miRNA。采用逆转录-定量聚合酶链反应(RT-qPCR)在独立队列(n=8)中验证差异表达的 miRNA。预测差异表达 miRNA 的靶基因,并进行基因本体论术语富集和京都基因与基因组百科全书通路分析。miRNA 微阵列分析显示,7 个 miRNA(miRs)显著上调(hsa-let-7b-5p、hsa-miR-223-3p、hsa-miR-4485、hsa-miR-4644、hsa-miR-4800-5p、hsa-miR-6510-5p 和 hsa-miR-765),3 个 miRNA 显著下调(hsa-miR-33b-3p、hsa-miR-4443 和 hsa-miR-4515),与健康对照组相比,在急性 KD 中差异有统计学意义。RT-qPCR 检测到 hsa-miR-223-3p 的表达水平与微阵列结果一致。共预测到 hsa-miR-223-3p 的 62 个靶基因。在急性 KD 中,共鉴定出 10 个差异表达的 miRNA,其中 hsa-miR-223-3p 通过 RT-qPCR 验证。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea2d/5780174/1a65d59bbd01/MMR-17-01-0932-g00.jpg

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