Identification Of Core Difference Expression Genes For Diabetic Foot Ulcers And Potential Molecular Pathology Mechanisms

Objective:Bioinformatics methods were used to analyze the gene expression spectrum of diabetic foot ulcers,diabetic foot skin,and non-diabetic skin differences,screen the associated differential expression genes(differentially expressed genes,DEGs)and their corresponding microRNAs(miRNAs),long-chain non-coding RNA(LncRNA),and verify the screening of gene expression in order to find potentially applicable clinical diagnostic and therapeutic targets.Method: GEO(http://www.ncbi.nlm.nih.gov/geo/)analyzes gene chip data sets with the R software's limma package package to screen diabetic foot ulcers,diabetic foot skin,non-diabetic skin-to-skin DEGs,miRNAs,and LNRNAs.Use DAVID bioinformatics software to annotate the functions of the Genetic Ontology,GO,Genes and Genomes of Kyoto Encyclopedia of Genes and Genomes.Integrate BioGRID and IntAct interaction information to find deGs interactions.Build a Protein Mutual Network(PPI)using String Online,and use biochart visualization tool Cytoscape software to build miRNA-DEGs,LncRNA-DEGs regulatory networks.Using RT-PCR to verify the bioinformatics analysis results,the expression levels of the core markers of diabetic foot ulcer tissue UBC,hsa-miR-148A-3p,hsa-miR-4516,hsa-miR-575,NR-024209,NR-036530.Results: The obtained chip data GSE80178,GSE84971,GSE68185,platform-based GPL16686,GPL17537,of which the experimental platform GPL16686 is Affymetrix's Affymet Humanrix Gene 2.0 ST Array chip,a total of 26 samples,identified overlapping upward gene expression gene expression with 29 differences It was found that the difference gene function enrichment of overlapping up was mainly concentrated in the functions of positive regulation of blood clotting,and the difference gene function of overlapping reduction was mainly enriched in the process of positive regulation of apoptosis process.The main enrichment in the biological process is in the process of changes in cell cycle,etc.Overlapping uplifted differences expressed miRNA,LncRNA have 20,5,overlapping downward differenceexpression expression miRNA,LncRNA have 7.Combined with the results of PPI and miRNA-DEG and LncRNA-DEG regulatory networks,UBC is at the core node position in the PPI network,and hsa-miR-148A-3p,hsa-miR-4516,hsa-miR-575 is located in the important position in the miRNA-DEGs regulatory network diagram.NR-024209,NR-036530 is at a critical location in the LncRNA-DEGs regulatory network diagram.RT-PCR validation shows that it is consistent with bioinformatics analysis and prediction trends.Conclusion: Our study,which screens differentially expressed genes,miRNA,and LncRNA through bioinformatics analysis,provides a new perspective on the molecular pathology mechanisms of exploring diabetic foot ulcers,which in turn can contribute to the prevention,treatment,and prognosis of diabetic foot ulcers.Our study shows that the heat shock protein 70(HSP70)family is closely related to DFU differential expression genes,the key gene UBC is an important marker of the evolution of DFU disease,and three miRNAs,two LncRNAs,may be potential diagnostic markers or therapeutic targets for DFU.The Wnt signaling pathway may be an important molecular pathological mechanism for the development of diabetic foot ulcers.