基于加权基因共表达网络分析运动性精子结构域包含蛋白2在类风湿关节炎中的表达及与疾病活动度的相关性

目的利用加权基因共表达网络分析(WGCNA)及实验验证寻找RA相关的关键基因。方法从GEO数据库下载了RA患者基因芯片数据,构建基因网络,利用WGCNA将基因划分为不同的模块,将与RA临床症状相关的模块中的关键基因进行了基因本体论分析。随后使用GEO不同的数据集用受试者工作特征曲线(ROC)评价关键基因对RA诊断的准确性。此外,还通过实时荧光定量反转录PCR(RT-PCR)及蛋白质印迹法验证关键基因在RA中的表达,分析其与DAS28的关系。采用配对样本t检验和Pearson相关性分析对结果进行分析。结果共筛选出5413个基因构建了加权基因共表达网络,将基因分为23个模块。其中,黑色模块与RA临床症状密切,包含346个基因。富集分析及京都基因与基因组百科全书(KEGG)信号通路分析显示其要富集于对IL-6的正调控、IL-1β分泌、破骨细胞分化、NOD样受体信号通路、辅助性T细胞(Th)17细胞分化等多个与RA密切相关的通路。其中运动性精子结构域包含蛋白2(MOSPD2)与临床症状具有明显相关性,在血液单核细胞、骨髓单核细胞中高表达(t=2.238,P=0.032;t=3.153,P=0.006),在RA关节滑膜液中与血液中表达呈正相关(r=0.683,P=0.03)。ROC曲线分析表明,MOSPD2能区分RA和对照组(曲线下面积分别为0.855和0.726)。RT-PCR及蛋白质印迹法结果显示,MOSPD2在RA患者中表达上调(t=-3.96,P=0.02)。MOSPD2在血液中的表达水平与RA患者的DAS28呈正相关(r=0.8846,P=0.0462)。结论MOSDP2与RA患者临床症状密切相关,可能是诊断及治疗RA的靶点之一。

Analysis on the expression and clinical significance of MOSPD2 in rheumatoid arthritis based on weighted gene co-expression network

Objective To identify the key genes related to rheumatoid arthritis(RA)by to the weighted gene co-expression network analysis(WGCNA)and experimental verification to find key genes related to RA.Methods The microarray data of RA were downloaded from the Gene Expression Omnibus(GEO)database.Gene network was constructed,and the genes were classified into different modules using WGCNA.HUB genes in modules related to RA clinical symptoms were analyzed by gene ontology.Subsequently,different data sets of GEO were used to verify the expression profile and diagnostic capacity of the HUB genereceiver operating characteristic curve(ROC)].In addition,the expression of HUB gene in RA was verified by real time polymerase chain reaction(RT-PCR)and Western blot,and the relationship between key genes and disease activity score 28 joints(DAS28)was analyzed.Paired-sample t-test and Pearson's correlation analysis was used for statistical analysis.Results A total of 5413 differentially expressed genes were filtered.Weighted gene coexpression network was constructed and genes were classified into 23 modules.Among them,the black module is closely related to the clinical symptoms of RA,which contained 346 genes.Enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)signal pathway analysis showed that it was to be enriched in the positive regulation of interleukin 6,interleukin 1 beta secretion,osteoclast differentiation,NOD-like receptor signaling pathway,T helper cell 17(Th17)cell differentiation and many other pathways closely related to RA.Motile sperm domain-containing protein 2(MOSPD2)was significantly correlated with clinical symptoms.It was highly expressed in blood monocytes and bone marrow monocytes(t=2.238,P=0.032;t=3.153,P=0.006),and positively correlated with blood expression in RA joint synovial fluid(r=0.683,P=0.03).ROC curve analysis determined that MOSPD2 could distinguish RA from the control group(the area under the curve was 0.855 and 0.726)respectively.RT-PCR and Western blotting results showed that MOSPD2 was up-regulated in RA patients(t=-3.96,P=0.02).MOSPD2 expression levels in blood were positively correlated with DAS28 in RA patients(r=0.8846,P=0.0462).Conclusion MOSDP2 is closely related to the clinical symptoms of RA patients,and may be one of the targets for the diagnosis and treatment of RA.