Bioinformatics Analysis Of Genes Related To Benign Airway Stenosis Induced By Artificial Airway

Objective: In this study,the gene data of geo database were analyzed in depth,and the results were verified in clinical samples,in order to find the hub gene of benign tracheal stenosis.Methods:1.Collect and process the data in GEO,divide the gene data set into GTS group and GCON group,and select a large number of DEGs through bioinformatics analysis.Go and KEGG were used to analyze DEGs.The results are processed by R language program and presented in the form of heat map and volcano plot.In addition,hub genes are screened out by constructing PPI network,important modules and hub gene network of DEGs,and diseases related to them are identified in CTD database.2.The venous blood of patients with artificial airway was collected in clinic.The blood samples of patients with benign tracheal stenosis were set as TS group,and the blood samples of patients without stenosis were set as con group.RNA was extracted from blood samples with RNAisoplus(Trizol)kit.RT-q PCR was performed on key genes in lightcycle ? 4800 system to analyze the expression of core genes in clinical samples.Finally,the correlation between the statistical results and the demographic characteristics and clinical manifestations of the patients was analyzed to identify the hub genes.Results: In the data set [GSE10936581(GPL16570 platform)],there was a strong correlation between the benign tracheal stenosis group(BTS group)and the normal group(CON group).A total of 194 differentially expressed genes(DEGs)were analyzed,of which 61 were down regulated and 133 up regulated.The results of GO analysis were significantly enriched in nuclear fission,cell cycle and cell division.According to KEGG analysis,p53 signaling pathway and cell cycle are the main pathways.Mki67,Ccnb1 and Ccnb2 are the hub genes of tracheal stenosis group in the data set,and these genes are identified to be related to tracheal diseases,cartilage diseases,tracheal tumors,fibroadenomas and connective tissue diseases.RT-q PCR analysis showed that Mki67(P<0.05),Ccnb1(P<0.05)and Ccnb2(P<0.05)were significantly up-regulated.It should be noted that in the above bioinformatics results,the expression of Mki67,Ccnb1 and Ccnb2 increased with the degree of stenosis in BTS group.There was a significant positive correlation between stenosis degree and Mki67,Ccnb1 and Ccnb2,and a strong positive correlation between TS degree and Mki67(R=0.906,P<0.001).The degree of TS was positively correlated with Ccnb1(R=0.953,P<0.001).The degree of TS was positively correlated with Ccnb2(R=0.958,P<0.001).The relative expression of Mki67 was positively correlated with Ccnb1(R=0.912,P<0.001).The relative expression of Mki67 was positively correlated with Ccnb2(R=0.862,P<0.001).The relative expression of Ccnb1 was positively correlated with Ccnb2(R=0.936,P<0.001).Through Spearman correlation analysis,Heatmap shows the strongly correlation among Mki67,Ccnb1 and Ccnb2.Conclusion:1.The occurrence and development of benign tracheal stenosis can not be separated from abnormal activation and expression of genes related to cell proliferation and cell cycle.2.Mki67,ccnb1 and ccnb2 genes are closely related to the occurrence of benign tracheal stenosis.