WT1-AS Competitively Combined With MiR-200a-3p To Regulate The Expression Of NAMPT And Affect The Molecular Mechanism Of Lung Adenocarcinoma

Objective: Using bioinformatics and medical statistical analysis methods,the sequencing data of lung adenocarcinoma gene expression in the human tumor genome database were analyzed,and patients were grouped based on the expression level of the key oncogene MYC,and the genes with abnormal expression levels were identified.A competitive endogenousRNA regulatory network composed of differentially expressed lncRNAs,miRNAs and mRNA was constructed.Combined with patients’ clinical data and follow-up information,genes closely related to the prognosis of patients were screened out.The experiment verified the function and mechanism of key lncRNAs in the development of lung adenocarcinoma through competitive endogenousRNA regulation mode.Research methods: 1.Download the gene expression sequencing data of lung adenocarcinoma patients and the corresponding patient clinical information and follow-up information from the TCGA database,and use the R(version 3.6.0)"edge.R" package to identify DElncRNA,DEmiRNA and DEmRNA.Use R "cluster Profiler" package and STRING to perform gene ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis and Construct a differential gene protein interaction network to identify key genes and signal pathways.The ceRNA regulatory network was constructed by cytoscape version 3.6.0,and the prognostic-related genes of lung adenocarcinoma were identified by single-factor and multi-factor Cox regression analysis,the research target lncRNA was determined,and its relationship with clinicopathological characteristics was analyzed;2.q RT-PCR detection of tissue The expression level of targetRNA in samples and cell lines,and combined with clinical information to verify its correlation with clinicopathological characteristics;3.Use molecular imprinting and other tests to verify and detect theexpression level of targetRNA-encoded protein in tissue samples and cell lines.Results: 1.Identified 1639 differentially expressed lncRNAs,99diffe-rentially expressed miRNAs and 2569 differentially expressed mRNAs,and constructed a ceRNA regulatory network containing 12 differentially expressed lncRNAs,39 differentially expressed miRN As and 62 differentially expressed mRNAs.It is found that 2lncRNAs,3 miRNAs and 6 mRNAs are closely related to the poor prognosisof patients with lung adenocarcinoma.Because the high expression level of LncRNA WT1-AS is significantly related to the shortened survival time and poor prognosis of patients,the WT1-AS/miR-200a-3p/NAMPT regulatory axis is selected as the research object,and the next step is to explore in depth.2.The expression levels of WT1-AS in lung epithelial bronchial cell lines and lung adenocarcinoma cell lines were detected by q RT-PCR,the expression levels of WT1-AS and NAMPT were verified in lung adenocarcinoma cell lines and lung adenocarcinoma pathological tissues.Significantly higher,and significantly related to clinical stage,pathological grade,lymph node metastasis(p<0.05).Conclusion: This study constructed a lung adenocarcinoma ceRNA regulatory network based on the expression level of the key oncogene MYC and screened the WT1-AS/miR-200a-3p/NAMPT regulatory pathway,which verified WT1-AS as a competitive endogenousRNA through experiments,Through competitive binding with miR-200a-3p up regulate NAMPT,promote the progression of lung adenocarcinoma,provide new ideas for the molecular mechanism of lung adenocarcinoma,and provide new potential targets for the diagnosisand treatment of lung adenocarcinoma.