Bioinformatics Analysis And Functional Studies Of Hsa-miR-26b-5p In Scleroderma

?Object?To identify differentially expressed genes(DEGs)and relative miro RNAs,and investigate their roles in the pathogenesis of scleroderma by using bioinformatics analysis.?Methods?We downloaded the microarray dataset GSE95065 from the Gene Expression Omnibus(GEO)database and then analyzed the data by using GEO2 R.The Database for Annotation,Visualization,and Integrated Discovery(DAVID)was used for functional pathway enrichment analyses of differentially expressed genes(DEGs),and Cytoscape software was used to generate the protein-protein interaction(PPI)network.In addition,Omics Net was used to predict the mi RNAs for the hub genes of scleroderma.The function of hsa-mi R-26b-5p was examined by transfecting hsa-mi R-26b-5p inhibitor into human skin fibroblasts.?Results?(1)After the analyses of GSE95065,770 DEGs were identified,they are unique genes in scleroderma skin tissue samples compared with normal skin samples.In addition,we identified 142 upregulated genes and 628 downregulated genes.(2)The GO term enrichment analysis indicated that the upregulated genes were mainly involved in immune response,while the downregulated genes were involved in signal transduction.the KEGG pathway analysis revealed that the upregulated genes took part in the cytokine-cytokine receptor interaction pathway,whereas the downregulated genes mainly participated in the neuroactive ligand-receptor interaction.(3)The 770 identified DEGs were uploaded to the STRING database to construct a PPI network(combined score > 0.9),and a total of 309 nodes and 819 edges were generated.The 22 highest-scoring nodes were defined as key genes.(4)All of the hub genes were uploaded to Omics Net,the results indicated that hsa-mi R-26b-5p might target CXCL9 and CXCL13,and that hsa-mi R-335-5p might target CXCL9,CCL19 and CCL25.(5)Hsa-mi R-26b-5p expression dramatically decreased after transfection with the hsa-mi R-26b-5p inhibitor.As a result,the decreased hsa-mi R-26b-5p expression led to decreased expression of ?-SMA?FAP?Col1A2?Col4A1 m RNA as well as CXCL9 ?CXCL13 m RNA compared to the mi R-NC group after TGF? stimulation.?Conclusions?(1)We successfully identified 770 DEGs of scleroderma and uplodeded them to DAVID database for functional and pathway enrichment analyses,which may provide new clues for better understanding on the molecular mechanism of scleroderma.(2)A PPI network was conducted and revealed that CXC motif chemokine ligand family members may take part in the occurrence and development of secleroderma.We also found that hsa-mi R-26b-5p might target CXCL9 and CXCL13.(3)Hsa-mi R-26b-5p downregulation might inhibit the fibrosis process by targeting CXCL9 and CXCL13,which might provide new insight into the diagnosis and treatment of scleroderma.