颈椎后纵韧带骨化症肥胖相关基因的筛选及鉴定

Screening and identification of obesity-related genes in cervical ossification of posterior longitudinal ligament

目的:探索颈椎后纵韧带骨化症(OPLL)肥胖相关基因及其功能特征。方法:从高通量基因表达(GEO)数据库中下载颈椎OPLL相关的转录组数据集GSE69787,与Gene数据库中肥胖相关基因进行筛选,获得颈椎OPLL肥胖相关差异表达基因(DEGs)。使用DAVID软件和R软件对肥胖相关DEGs进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析,应用Cytoscape软件将数据可视化,并通过cytoHubba工具包计算得到中心基因。采用实时定量PCR(qRT-PCR)检测中心基因在颈椎后纵韧带组织中的表达水平。结果:通过GSE69787数据集与肥胖相关基因筛选得到24个颈椎OPLL肥胖相关DEGs,构建并计算这24个肥胖相关DEGs的蛋白质相互作用网络,最终筛选出10个中心基因。GO/KEGG富集分析结果显示这10个肥胖相关DEGs主要富集于蛋白激酶B信号通路及其调控、骨化、炎症反应调控、骨矿化及PI3K-Akt信号通路、MAPK信号通路等生物学过程和生物信号通路。最后通过qRT-PCR验证发现LEP、IGF2、IL18、BMP2和LOX在颈椎OPLL组韧带组织中高表达(P均<0.05),MMP9、PTGS2在颈椎OPLL组韧带组织中低表达。结论:LEP、IGF2、IL18、BMP2、LOX、MMP9、PTGS2是颈椎OPLL患者与肥胖相关的基因,其异常表达可能与颈椎OPLL的发生、发展相关,并可能为合并肥胖的颈椎OPLL患者的诊断和治疗提供新的思路。

Objective:To explore the differentially expressed obesity-related genes and their function in the cervical ossification of posterior longitudinal ligament(OPLL).Methods:The OPLL-related transcriptome dataset GSE 69787 was downloaded from the Gene Expression Omnibus(GEO)database,and obesity-related genes were screened in the Gene database to obtain obesity-related differentially expressed genes(DEGs)in OPLL.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses obesity-related DEGs were performed using DAVID software and R software.The data were visualized using Cytoscape software,and hub genes were calculated by cytoHubba tool.Quantitative real-time PCR(qRT-PCR)was used to detect the expression levels of hub genes in posterior longitudinal ligament tissue.Results:Twenty-four OPLL obesity-related DEGs were obtained by screening GSE 69787 and obesity-related genes.A protein-protein interaction network of these 24 obesity-related DEGs was constructed and calculated,and finally,10 important hub genes were selected.GO/KEGG enrichment analyses indicated that these 10 obesity-related DEGs were mainly enriched in biological processes and signaling pathways such as protein kinase B signaling pathway and its regulation,ossification,regulation of inflammatory response,bone mineralization,PI 3 K-Akt signaling pathway and MAPK signaling pathway.Finally,qRT-PCR verified that LEP,IGF 2,IL 18,BMP 2,and LOX were highly expressed in OPLL ligament tissue(P<0.05),while MMP 9 and PTGS 2 were low expressed in OPLL ligament tissue(P<0.05).Conclusions:LEP,IGF 2,IL 18,BMP 2,LOX,MMP 9,and PTGS 2 are genes associated with obesity in OPLL patients.The abnormal expression of these genes may be related to the occurrence and development of OPLL,providing new ideas for the diagnosis and treatment of OPLL patients with obesity.