| Objective: The purpose of this study is to analyze the differential expression of lncRNAs and mRNAs in serum rheumatoid arthritis(RA)patients.Further study its function in target cells and preliminarily explore the role of lncRNAs in the occurrence and development of RA.Methods: Based on the gene chip technology,the preliminary screening of the differential expression of lncRNAs and mRNAs in the mixed serum of RA patients and healthy controls.And we verified by real-time fluorescence quantitative PCR(qRTPCR).Through correlation analysis,ROC curve and logistic regression to analyze the relationship between abnormal expression of lncRNAs and laboratory markers.GO and KEGG pathways were analyzed,and the co-expression network of lncRNA and mRNA was constructed.Two data sets in GEO database,GSE101193 and GSE94519,were mined by R Studio,and the intersection was obtained with the two verified lncRNAs.RA-FLS were cultured and lentiviral vector overexpressed lncRNA XR 428141.1 was constructed.The proliferation,invasion,migration and apoptosis of RA-FLS were further studied by lentiviral vector infection.Results: 1.The differential expression profile was analyzed by gene chip technology.The results showed that there are 1693 lncRNAs and 1541 mRNAs with significant differential expression.QRT-PCR verified that lncRNA XR＿428141.1 was up-regulated in serum of RA patients(P=0.0075)and lncRNA ENST00000478103.2was down-regulated(P=0.0023),which was consistent with chip results.In RA group,the ROC(AUC)of lncRNA XR＿428141.1 and ENST00000478103.2 were 0.872 and0.837,respectively.Correlation analysis showed that lncRNA XR＿428141.1 was positively correlated with RF and anti-CCP,while lncRNA ENST00000478103.2 was negatively correlated with RF and anti-CCP.Binary logistic regression analysis showed that lncRNA XR＿428141.1 could be used as a predictor of RA with a risk ratio of 1.941 and 95% confidence interval of 1.140-3.305.By binary logistic regression analysis,the fitting degree of the model was 93.1%.2.GO&KEGG pathway analysis shows that 376 up-regulated lncRNAs are involved in 1455 biological processes,284 molecular functions,and 253 cell compositions.KEGG pathway analysis may be mainly in signal transduction,immune disease,metabolism,immune system and endocrine system pathways play a role.LncRNA XR＿428241.1 and ENST00000478103.2 have a co-expression relationship with mRNA SNX21 and UNC93B1.According to the mining analysis of two data sets in the GEO database: GSE101193 and GSE94519,we performed two intersection verifications,but the intersection results were not obtained.It is confirmed that lncRNA XR＿428141.1 and ENST00000478103.2 do not overlap with the two data sets.3.Transwell cell invasion assay,the results showed that by overexpressing lncRNA XR＿428141.1 could significantly promote RA-FLS invasion compared with the control group.The results of CCK-8 assay showed that lncRNA XR＿428141.1could further promote the proliferation of RA-FLS.Based on the apoptosis assay,it is predicted that lncRNA XR＿428141.1 may have the function of inhibiting RA-FLS apoptosis.Conclusions: LncRNA XR＿428141 and ENST00000478103 may be involved in the physiological and pathological process of RA,which has potential application value for the clinical diagnosis of RA and provides theoretical basis for the clinical treatment of RA. |
