Clinical Significance Of Rhythm Gene NPAS2 In Lung Adenocarcinoma

BackgroundLung cancer has been one of the leading causes of cancer-related death for decades.The incidence of lung adenocarcinoma is increasing and is the most common pathological type of lung cancer.Although lung adenocarcinoma currently has various treatment methods such as surgery,radiotherapy,targeted therapy,immunotherapy,and chemotherapy,the incidence and mortality of lung cancer remain high.Therefore,the pathogenesis of lung adenocarcinoma needs to be further clarified and explored.Humans are diurnal creatures,active during the day and sleeping at night.The endogenous circadian system controls this rhythmic activity and physiological oscillations.Disruption of endogenous circadian balance increases the risk of many diseases,including psychiatric disorders,cardiovascular problems,metabolic disorders,aging,immunodeficiency,reproductive and neuroendocrine dysfunction,and cancer.As a core circadian gene,NPAS2 is involved in a variety of cancer-related processes and has been shown to have prognostic significance in a variety of cancers.The effect and mechanism of NPAS2 gene on lung adenocarcinoma are still unclear.The clinical significance of NPAS2 gene in lung adenocarcinoma was explored through the verification of bioinformatics methods and clinical samples.ObjectiveTo explore the expression of NPAS2 gene in lung adenocarcinoma and its association with related clinical parameters,to analyze its effect on the overall survival of patients,and to explore the expression of NPAS2 in lung adenocarcinoma.Predictive value in cancer prognosis.Methods1.We download the LUAD transcriptome sequencing and clinical data data from The Cancer Genome Atlas(TCGA),and analyze the expression difference of NPAS2 in LUAD tumor tissue and normal lung tissue.2.Western blot was used by us to detect the expression of NPAS2 protein in 12 fresh lung adenocarcinoma tissues and the corresponding adjacent normal tissues,then,the differences in expression were analyzed.3.RT-q PCR was used by us to detect the expression of NPAS2 m RNA in the collected12 fresh lung adenocarcinoma tissues and the corresponding adjacent normal tissues,and to analyze the difference in expression between the two groups.4.We analyze the association of NPAS2 m RNA expression in TCGA database with tumor stage and overall survival in LUAD patients.Immunohistochemical staining was used to detect the expression of NPAS2 protein in 124 lung adenocarcinoma samples collected,and analyzed the association of NPAS2 protein expression with the clinical characteristics and overall survival of patients with lung adenocarcinoma.5.Using the data in the TCGA database,the potential carcinogenic mechanism of NPAS2 was mined by gene set enrichment analysis(GSEA).Results1.The data analysis results of TCGA database show that compared with normal lung tissue,the expression of NPAS2 m RNA was up-regulated in LUAD tissue,and the difference is statistically significant(P <0.05).2.12 Western blot results of fresh lung adenocarcinoma and adjacent normal tissue show that compared with normal lung tissue,the expression of NPAS2 protein in lung adenocarcinoma is significantly increased,and the difference is statistically significant(P<0.05).3.12 RT-q PCR results of lung adenocarcinoma and adjacent normal tissue show that compared with normal lung tissue,the expression of NPAS2 m RNA in lung adenocarcinoma is significantly increased,and the difference was statistically significant(P <0.05).4.The data analysis results of TCGA database show that compared with T1-2 group,the proportion of high NPAS2 m RNA expression in T3-4 group is higher,and the difference is statistically significant(P <0.05);compared with patients with low NPAS2 expression,high NPAS2 m RNA The overall survival time of patients with expression is shorter,and the difference is statistically significant(P <0.05).The results of immunohistochemical staining show that compared with the T1-2 group,the expression of NPAS2 protein in the T3-4 group is higher,and the difference is statistically significant(P<0.05).The expression of NPAS2 protein is higher in the middle period,and the difference is statistically significant(P <0.05).Compared with stage II,the expression of NPAS2 protein is higher in stage III,and the difference is statistically significant(P <0.05).Compared with patients with low NPAS2 expression,patients with high NPAS2 expression has a shorter overall survival time(P <0.05).5.GSEA gene enrichment results show that high expression of NPAS2 can induce apoptosis,ERBB signaling pathway,leukocyte transendothelial migration,cancer pathway,small cell lung cancer,arrhythmogenic right ventricular cardiomyopathy,axon guidance,dilated cardiomyopathy,dorsal-ventral axis formation,hypertrophic cardiomyopathy,adherens junctions,ECM receptor interactions,focal adhesions,phosphatidylinositol signaling system,regulation of cytoskeletal action,glycosaminoglycan biosynthesis of chondroitin sulfate,Biosynthesis of glycosphingolipids-lactate and new latex series,O-glycan biosynthesis pathway activation.ConclusionNPAS2 is highly expressed in LUAD and is associated with local tumor invasion,TNM staging and overall survival,and can be used as an independent prognostic predictor in LUAD.