Expression Of AREG And Associated Analysis In Squamous Cell Carcinoma Of The Lung

Objective: To investigate the expression of amphiregulin(AREG)in lung squamous-cell carcinoma(Lung Squamous-cell Carcinoma,LUSC),and to determine the prognostic effect of AREG expression on LUSC and its related mechanism through bioinformatics analysis,in order to provide Clinical decision-making and risk management provide a solid scientific basis.Methods: In this study,the corresponding clinical information and gene expression profiles of LUSC patients were downloaded from The Cancer Genome Atlas(TCGA)database and the Gene Expression Omnibus database(GEO),and the LUSC patients and normal controls were extracted.The expression level of AREG in the population was analyzed,and the correlation between the expression level of AREG and the clinical parameters of LUSC patients was analyzed using the "stats" and "Car" R packages,and the relationship between the expression level of AREG and the clinical characteristics of LUSC patients was evaluated by logistic regression analysis.Then,we divided the survival data of LUSC patients in the TCGA database into AREG high expression group and AREG low expression group according to the median expression value of AREG,and performed Kaplan-Meier analysis using "survival" R package,and adopted univariate and multivariate Cox analysis was used to evaluate the correlation between clinical parameters of LUSC patients and overall survival(OS),disease specific survival(DSS)and progression-free interval(PFI).Further,we used the TIMER database to explore the correlation of AREG copy number variation with the abundance of 6 TIICs(including B cells,CD4 T cells,CD8 T cells,neutrophils,macrophages and dendritic cells),and Gene ontology(GO)and Kyoto Encyclopedia of Genes and Gnomes(KEGG)were used to explore the signaling pathways related to the role of AREG.Finally,we used "Spearmam" to analyze the correlation of AREG expression with immune checkpoint-related genes including CTLA4,PDL-1,LAG3,and IDO1.On the other hand,we clinically collected serum from 33 LUSC patients and30 healthy people,and used enzyme-linked immunoassay(ELISA)method to detect the content of AREG in serum samples,and analyzed AREG in serum The correlation between the expression level of LUSC and the clinical parameters of LUSC patients was verified by some results of bioinformatics analysis.Results:(1)Analyses in different databases showed that there were differences in the expression levels of AREG between lung tissue and normal tissue of LUSC patients.(2)The serum of LUSC patients and healthy people were collected and analyzed clinically.The results showed that the expression of AREG in the blood of LUSC patients was significantly higher than that of healthy people.(3)Through the analysis of TCGA database and clinical data,we found that the expression level of AREG was not significantly different from T stage,N stage,M stage,age,gender,and smoking.(4)Survival analysis results showed that patients with high AREG expression were associated with shorter PFI(P = 0.039)and DSS(P = 0.01),but not OS(P =0.111).(5)Multivariate analysis showed that AREG was an independent risk factor for predicting DSS and PFI,but not for OS.Pathological stage was an independent predictor of OS,DSS and PFI.Age and gender were not independent predictors of OS,DSS and PFI.(6)Using TIMER database to analyze the relationship between AREG expression and immune infiltrating cells,the low expression group had more CD8 T cells(P < 0.01),dendritic cells(P < 0.001),macrophages(P < 0.001),neutral Granulocytes(P < 0.001),CD56 natural killer cells(P < 0.01),mast cells(P < 0.001),immature dendritic cells(P < 0.001),eosinophils(P < 0.001),Th1 cells(P < 0.001),Th2 cell(P < 0.05)infiltration.In addition,AREG expression was found to be significantly negatively correlated with tumor purity(R =-0.265,P = 3.84e-09),B cell infiltration level(R =-0.157,P = 6.32e-04),and AREG expression was correlated with macrophage There was a significant positive correlation between cells(R = 0.245,P =6.06e-08)and neutrophil infiltration(R = 0.214,P = 2.41e-06).(7)According to GO and KEGG enrichment scores,AREG may be related to cell proliferation,apoptosis,and immune response signaling pathways.(8)The results showed that there was a positive correlation between the expression level of AREG and CTLA-4.Conclusion:(1)There are differences in the expression level of AREG between LUSC patients and normal controls;(2)AREG can be used as an independent prognostic indicator of DSS and PFI in LUSC;(3)The expression level of AREG is related to immune infiltration;(4)AREG may be a potential therapeutic target for LUSC.