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运用综合生物信息学分析和实验验证鉴定Slc6a19os和SOX11为神经性疼痛中的两个新型必需基因

Identification of Slc6a19os and SOX11 as Two Novel Essential Genes in Neuropathic Pain Using Integrated Bioinformatic Analysis and Experimental Verification.

作者信息

Chen Peng, Wang Chen, Lin Dongsheng, Li Bing, Ye Shuai, Qu Jinglian, Wang Wenjing

机构信息

Basic Medical School, Guizhou University of Traditional Chinese Medicine, Guiyang, China.

First Clinical Medical School, Guangzhou University of Chinese Medicine, Guangzhou, China.

出版信息

Front Neurosci. 2021 Jan 28;15:627945. doi: 10.3389/fnins.2021.627945. eCollection 2021.

Abstract

The aim of this study was to identify critical genes associated with neuropathic pain. We also used the competing endogenous RNA (ceRNA) hypothesis to identify related long non-coding RNAs (lncRNAs) and messenger RNAs (miRNAs) with potential regulatory roles. We downloaded GSE107180 from the Gene Expression Omnibus database, screened differentially expressed genes (DEGs) using R software, performed comprehensive bioinformatic analyses, and validated the expression of lncRNA Slc6a19os, miR-125a-5p, miR-125b-5p, miR-351-5p, and Sox11 by qRT-PCR and Western blots. We identified 620 DEGs in spared nerve injury (SNI) mice compared with sham (control) mice, including 309 mRNAs and 311 non-coding RNAs. The up-regulated mRNAs were enriched primarily in several inflammation-related GO biological processes and KEGG signaling pathways. A ceRNA network was constructed that included 82 mRNAs, 4 miRNAs, and 2 lnRNAs. An ingenuity pathway analysis (IPA)-based interaction network for mRNAs differentially expressed in the ceRNA identified several biological processes, including "cellular development, connective tissue development and function, tissue development." Compared with sham mice, lncRNA Slc6a19os and Sox11 expression were significantly up-regulated in dorsal root ganglion (DRG) samples from SNI mice detected using qRT-PCR and Western blots ( < 0.05). MiR-125a-5p, miR-125b-5p, and miR-351-5p expression were down-regulated in DRG samples from SNI mice detected using qRT-PCR ( < 0.05). We concluded that Sox11 and lncRNA Slc6a19os were novel essential genes in the pathogenesis and progression of neuropathic pain and speculated that these two genes were regulated by miR-125a-5p, miR-125b-5p, and miR-351-5p.

摘要

本研究的目的是鉴定与神经性疼痛相关的关键基因。我们还利用竞争性内源性RNA(ceRNA)假说,来鉴定具有潜在调控作用的相关长链非编码RNA(lncRNA)和信使RNA(miRNA)。我们从基因表达综合数据库下载了GSE107180,使用R软件筛选差异表达基因(DEG),进行全面的生物信息学分析,并通过qRT-PCR和蛋白质印迹法验证lncRNA Slc6a19os、miR-125a-5p、miR-125b-5p、miR-351-5p和Sox11的表达。与假手术(对照)小鼠相比,我们在 spared nerve injury(SNI)小鼠中鉴定出620个DEG,包括309个mRNA和311个非编码RNA。上调的mRNA主要富集于几个与炎症相关的GO生物学过程和KEGG信号通路。构建了一个ceRNA网络,其中包括82个mRNA、4个miRNA和2个lnRNA。基于 Ingenuity Pathway Analysis(IPA)的ceRNA中差异表达mRNA的相互作用网络,确定了几个生物学过程,包括“细胞发育、结缔组织发育和功能、组织发育”。与假手术小鼠相比,使用qRT-PCR和蛋白质印迹法检测发现,SNI小鼠背根神经节(DRG)样本中lncRNA Slc6a19os和Sox11的表达显著上调(<0.05)。使用qRT-PCR检测发现,SNI小鼠DRG样本中miR-125a-5p、miR-125b-5p和miR-351-5p的表达下调(<0.05)。我们得出结论,Sox11和lncRNA Slc6a19os是神经性疼痛发病机制和进展中的新的关键基因,并推测这两个基因受miR-125a-5p、miR-125b-5p和miR-351-5p的调控。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/baef/7876402/1ef740335228/fnins-15-627945-g0001.jpg

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