Exploratory Study On Differential Expression Profiles And Regulatory Network Of MRNA,miRNA And LncRNA In Type 2 Diabetes

Objective:In this study,high-throughput sequencing and bioinformatics analysis techniques were used to construct differential expression profiles of peripheral blood mRNA,miRNA and lncRNA in patients with type 2 diabetes,and based on the correlation between lncRNA-miRNA and miRNA-mRNA.The lncRNA-miRNA-mRNA regulatory network,which locates key genes in the network,clarifies the regulation of upstream and downstream genes,and provides a theoretical basis for studying the transcription and post-transcriptional levels of the development,diagnosis and treatment of type 2 diabetes.Methods:This study included 3 new cases of 40-60 year old male type 2 diabetes patients from the Second Hospital of Jilin University from August 2017 to June 2018.Three healthy and physically matched patients were included in the study.Group,venous blood was collected on an empty stomach and clinical data and related examination information were collected.The Illumina Hiseq sequencing platform was used for sequencing using the PE150 sequencing strategy.The genes were screened according to differential multiples and significant levels,and the screening conditions were differential expression multiples > 4 and P values < 0.05,and the distribution of differential genes was displayed using volcano maps and heat maps.The lncRNA-miRNA-mRNA regulatory network was constructed based on the correlation between lncRNA,miRNA and mRNA.Visualize the regulatory network using Cytoscape 3.7.0 software.After analyzing the network,the degree of centrality,proximity and centrality are obtained,and the key nodes of the network are selected with the three indicators being greater than or equal to the 95 th percentile of the population.The STRING online database was used to analyze the protein-protein interaction network?PPI?of genes in the lncRNA-miRNA-mRNA regulatory network.The PPI network was clustered using the MCODE plug-in of Cytoscape software.GO and KEGG enrichment analysis was performed using KOBAS 3.0.Results:1.According to the screening criteria,a total of 2,917 differentially expressed mRNAs?of which 1155 were up-regulated and 1762 were down-regulated?,33 differentially expressed miRNAs?27 of which were up-regulated,6 down-regulated?and 440 differentially expressed lncRNAs were obtained.?of which 366 are raised and 74 are lowered?.2.The lncRNA-miRNA-mRNA regulatory network was constructed based on the correlation between genes.The network includes 233 lncRNAs?up to 185,down-regulated by 48?,16 miRNAs?up-regulated by 14 and down-regulated by 2?and 139 mRNAs?Upgraded 76,down 63?.3.The main pathways of gene enrichment in the regulatory network of lncRNA-microNA-RNA include: Fcgamma-R-mediated phagocytosis,MAPK signaling pathway,T cell receptor signaling pathway,tuberculosis,lysosome,endocytosis,PI3K-Akt signaling pathway,p53 signaling pathway,other glycan degradation,mTOR signaling pathway,phagosome,B cell receptor signaling pathway.4.According to the criteria,10 key nodes in the regulatory network were screened out,including three lncRNA?lncRNA CTD-2542L18.1,lncRNA MIAT and LNC₀00322?,and seven microRNAs?hsa-mir-1271-5p,hsa-mir-3158-3p,hsa-mir-3158-5p,hsa-mir-330-5p,hsa-mir-5690,hsa-mir-6806-3p and hsa-mir-6842-3p?.5.lncRNA MIAT mainly regulates other glycan degradation,Fc?R-mediated phagocytosis,sphingolipid signaling pathway,apoptosis,phospholipase D signaling pathway,MAPK signaling pathway,tuberculosis and Rap1 signaling pathway.6.lncRNA CTD-2542L18.1 mainly regulates other glycan degradation,apoptosis,p53 signaling pathway,Fc?R-mediated phagocytosis,MAPK signaling pathway and PI3K-Akt signaling pathway.7.LNC₀00322 mainly regulates other glycan degradation,p53 signaling pathway,mRNA monitoring pathway,Fc?R-mediated phagocytosis and PI3K-Akt signaling pathway.8.Three network modules were identified from the PPI network ITGB2-ATP6V0C-TOM1,AKT2-MLST8-TSC2,and BECN1-ERBB2-BCL2L11.9.The module of ITGB2-ATP6V0C-TOM1 was mainly enriched in microbial infections signaling pathways.The module of BECN1-ERBB2-BCL2L11 was mainly enriched in cancer-related signaling pathways.The module of AKT2-MLST8-TSC2 was primarily enriched in endocrine and metabolic related signaling pathways.Conclusion:1.The mRNA in the regulatory network is mainly enriched in Fc?R-mediated phagocytosis,MAPK signaling pathway,endocytosis,PI3K-Akt signaling pathway,mTOR signaling pathway,apoptosis and AMPK signaling pathway.2.lncRNA MIAT,lncRNA CTD-2542L18.1 and LNC₀00322 are the key nodes in the construction of the regulation network.3.The module of AKT2-MLST8-TSC2 was primarily enriched in endocrine and metabolic related signaling pathways.The module may play an important role in the occurrence and development of type 2 diabetes.