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lncRNA-miRNA 网络分析揭示了 lncRNA NEAT1 和 KCNQ1OT1 在多发性硬化症中的生物标志物潜力。

LncRNA-miRNA network analysis across the Th17 cell line reveals biomarker potency of lncRNA NEAT1 and KCNQ1OT1 in multiple sclerosis.

机构信息

Department of Medical Genetics, Faculty of Medicine, Hormozgan University of Medical Sciences, Bandar Abbas, Iran.

Institute of Biotechnology, Shiraz University, Shiraz, Iran.

出版信息

J Cell Mol Med. 2022 Apr;26(8):2351-2362. doi: 10.1111/jcmm.17256. Epub 2022 Mar 10.

Abstract

Differentiation of CD T cells into Th17 cells is an important factor in the onset and progression of multiple sclerosis (MS) and Th17/Treg imbalance. Little is known about the role of lncRNAs in the differentiation of CD cells from Th17 cells. This study aimed to analyse the lncRNA-miRNAs network involved in MS disease and its role in the differentiation of Th17 cells. The lncRNAs in Th17 differentiation were obtained from GSE66261 using the GEO datasets. Differential expression of lncRNAs in Th17 primary cells compared to Th17 effector cells was investigated by RNA-seq analysis. Next, the most highlighted lncRNAs in autoimmune diseases were downloaded from the lncRNAs disease database, and the most critical miRNA was extracted by literature search. Then, the lncRNA-miRNA interaction was achieved by the Starbase database, and the ceRNA network was designed by Cytoscape. Finally, using the CytoHubba application, two hub lncRNAs with the most interactions with miRNAs were identified by the MCODE plug-in. The expression level of genes was measured by qPCR, and the plasma level of cytokines was analysed by ELISA kits. The results showed an increase in the expression of NEAT1, KCNQ1OT1 and RORC and a decrease in the expression of FOXP3. In plasma, an upregulation of IL17 and a downregulation of TGFB inflammatory cytokines were detected. The dysregulated expression of these genes could be attributed to relapsing-remitting MS (RR-MS) patients and help us understand MS pathogenesis better.

摘要

CD T 细胞向 Th17 细胞的分化是多发性硬化症 (MS) 发病和进展以及 Th17/Treg 失衡的重要因素。lncRNAs 在 CD 细胞向 Th17 细胞分化中的作用知之甚少。本研究旨在分析参与 MS 疾病的 lncRNA-miRNA 网络及其在 Th17 细胞分化中的作用。使用 GEO 数据集从 GSE66261 中获得 Th17 分化中的 lncRNAs。通过 RNA-seq 分析研究 Th17 原代细胞与 Th17 效应细胞相比 lncRNAs 的差异表达。接下来,从 lncRNAs 疾病数据库中下载自身免疫性疾病中最突出的 lncRNAs,并通过文献检索提取最关键的 miRNA。然后,通过 Starbase 数据库实现 lncRNA-miRNA 相互作用,并通过 Cytoscape 设计 ceRNA 网络。最后,使用 CytoHubba 应用程序,通过 MCODE 插件识别与 miRNA 相互作用最多的两个 hub lncRNAs。通过 qPCR 测量基因的表达水平,并通过 ELISA 试剂盒分析细胞因子的血浆水平。结果表明,NEAT1、KCNQ1OT1 和 RORC 的表达增加,FOXP3 的表达减少。在血浆中,检测到促炎细胞因子 IL17 的上调和 TGFB 的下调。这些基因的失调表达可能归因于复发缓解型多发性硬化症 (RR-MS) 患者,有助于我们更好地了解 MS 的发病机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d22/8995444/ff74b44fc7d4/JCMM-26-2351-g004.jpg

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