| BackgroundLung cancer is now one of the most serious threats to human health.In the statistics of lung cancer in China,the incidence of lung cancer and death ranks the first in men,the incidence of lung cancer in women is second only to breast cancer,and the death rate is still the first.NSCLC patients account for approximately 80% to 85% of all lung cancer incidence.The most common types of NSCLC are squamous cell carcinoma(SCC)and adenocarcinoma(AD).The most common type of lung cancer among nonsmokers is adenocarcinoma of the lung.At present,surgical resection of diseased tissues is the most effective method for the treatment of lung cancer.Although the treatment effect for lung cancer patients has been improved due to the improvement of medical conditions,surgical techniques and the upgrading of auxiliary equipment,the overall survival rate of NSCLC patients after 5 years is still 15% to 45%.The prognostic markers of tumor can affect and predict the survival outcome of cancer patients.Epidermal growth factor receptor(EGFR)belong to the Erb B family of receptor tyrosine kinases(RTKs),in the epithelial cells play an important role in physiological function,the activation of EGFR mainly mediated gene amplification and autocrine stimulation,is the main factor of the tumor is also the main factors of invasive cancer cells increased,on the other hand EGFR mutation and gene rearrangement often cause the change of receptor cells swallowing function,help to increase the signal conduction.The overall EGFR expression rate in NSCLC was 51%,and the poor prognosis of 70% of relapse-free patients with head and neck cancer,ovarian cancer,cervical cancer,esophageal cancer,and bladder cancer was associated with overexpression of EGFR.This topic through TCGA database download NSCLC patients with gene expression data,using Activeperl(perl execution tools),R packages and EGFR OSluca site analysis in patients with NSCLC tissue adjacent to carcinoma and the expression difference in tumor tissue and clinical information,and the relationship between the pathologic features,overall survival,at the same time get enrichment in GSEA enrichment analysis of EGFR high and low expression group related signaling pathways,The difference of EGFR expression between normal tissues and NSCLC tumor tissues was analyzed by Oncomine database for a second time and the results were visualized.Then the results were verified by immunohistochemistry(IHC)experiments using NSCLC tissue chip sections made independently.ObjectiveTo investigate the expression differences of EGFR in tumor tissues and adjacent tissues of NSCLC,and the relationship between EGFR and overall survival,clinical information and pathological characteristics of NSCLC patients.Methods1.Bioinformatics data analysisFirst download gene expression in NSCLC raw data from the TCGA database,use perl enforcement tools to download the data file and matedata combined genetic name translation again,forming a samples containing gene name and the name of matrix file and application of matrix R package quantity of single gene expression of EGFR in extraction,analysis its expression in the tissue and tumor tissue adjacent to carcinoma and EGFR expression quantity and quantity differences and relations between the patients' clinical information,pathological features.Then the expression data files and phenotypic data files in the gene matrix files were extracted for KEGG pathway enrichment analysis through GSEA software.Enter the gene name on the OSluca website,select the GSE19188 data set to analyze the relationship between EGFR expression and the overall survival of NSCLC patients,log on to the Oncomine database website,select the type of cancer as lung cancer,perform a secondary analysis on the expression of EGFR in normal tissues and tumor tissues,and generate a visual graph.2.Experimental verificationFrom January 2013 to December 2018,202 NSCLC patients admitted to our hospital for surgery were retrieved from the inpatient system of Huai He hospital of He Nan university,and their histological wax blocks were collected.Two NSCLC tissue chips were made by microchip machine,including 202 cases of tumor tissue and 113 cases of para-cancerous tissue.Then,the differences of EGFR expression among patients with different OS were observed through IHC experiment.SPSS software was used to analyze the differences of EGFR expression in different tissue types,and the relationship between different EGFR expression levels and clinical information and pathological characteristics was also analyzed.To verify the differences in the expression of EGFR in paracancer tissues and tumor tissues,as well as the relationship between the expression of EGFR and the OS,clinical and pathological characteristics of NSCLC.Results1.The data analysis of TCGA and Oncomine database showed that the expression of EGFR in tumor tissues was significantly different from that in adjacent tissues(P<0.05).The expression of EGFR in tumor tissues was high,while the expression of EGFR in adjacent tissues was relatively low.There were no differences in tumor size,lymph node metastasis,gender,age and stage(P>0.05).2.The results of OSluca survival analysis showed that the OSluca website was used to analyze the survival of the GSE19188 data set,and the results showed that there was a significant difference in OS between the high-expression EGRF group and the low-expression EGRF group(P < 0.05).3.Through GSEA analysis showed that the gene enrichment occurred in high EGFR expression channel mainly has three,respectively is: ERBB signaling pathways,WNT signaling pathways,and adherens junction occurred in the access of the low EGFR expression group a total of 35 major role in amino acid metabolism,and alzheimer's disease,type 1 diabetes,systemic lupus erythematosus,huntington's disease related pathways.4.IHC analysis of experimental results show that using the TMA technology and IHC to detect the expression of EGFR in NSCLC and prognosis and the relationship between the experimental results with the bioinformatics data analysis results and the previous traditional the IHC has consistent,namely EGFR expression in tumor tissue and tissue adjacent to carcinoma have significant differences,to observe the expression of EGFR in tumor tissue expression rate is 50.6%,and in the tissue adjacent to carcinoma of the EGFR expression rate was 2.3%(P < 0.05).There was no significant difference between EGFR expression and tumor size,lymph node metastasis,gender and age(P> 0.05).5.Kaplan-meier survival analysis showed that there were significant differences in OS between NSCLC patients with high and low EGFR expression(P<0.05).Conclusion1.There was a significant difference in the expression of EGFR in NSCLC tumor tissues and adjacent tissues.The expression of EGFR in adjacent tissues was generally low,while the expression of EGFR in tumor tissues was high.However,there was no difference in the expression of EGFR in different tumor sizes,lymph node metastasis,gender,age and stage.2.High expression of EGFR is associated with poor prognosis in NSCLC patients.3.Using TMA method to conduct IHC experiment can greatly improve the experimental efficiency and the results have higher accuracy or can replace the traditional IHC experiment method. |
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