Analysis Of Differentially Expressed Proteome In Urine From Lung Adenocarcinoma Patients

Objective:Lung cancer is the most deadly cancer in the world,and lung adenocarcinoma is the most common pathological type of lung cancer,accounting for about 40% of the total number of lung cancer.The cure rate of early lung cancer is high,but due to the atypical clinical manifestations of lung cancer,the sensitivity and specificity diagnostic biomarkers are limited.Most patients have advanced diagnosis at the late period,and the prognosis is poor.The 5-year survival rate is no more than 18%.Therefore,the discovery of efficient early diagnosis method is the key to effective treatment of lung cancer,prolong the life of patients.The development of proteomics provides a new means for the discovery of new tumor markers.Urine can not only reflect the relevant situation of the non-urinary system,as the experimental material is non-invasive,convenient and other advantages.In this study,urine proteome expression in healthy people and lung adenocarcinoma patients was analyzed and compared to identify diagnostic biomarkers associated with lung adenocarcinoma patients.Method:In this study,urine samples from lung adenocarcinoma group and healthy subjects were separated by ultra-high speed centrifugal precipitation method to obtain urine protein,and then the urine protein obtained by centrifugation was separated and grouped by gel electrophoresis.Decolorization,reduction and alkylation of the gel strip,the selection of target tape digestion and extraction.Protein and peptide compounds were obtained and identified by MS-Thermo-OrbiTrap-Velos mass spectr umetry analyzer.The obtained data were analyzed and identified by Uniprot database.GO analysis was conducted using DAVID biological information database,and the KEGG PATHWAY was used to screen out differential proteins related to immune and coagulation cascade pathways.Differential proteins related to tumor were selected and the clinical value of differential proteins was evaluated by ROC curve analysis and AUC.Finally,urine samples from patients with lung adenocarcinoma and healthy people were selected for ELISA detection to determine the expression of selectedlung adenocarcinoma biomarkers in lung adenocarcinoma and healthy people,so as to find out the biological markers related to lung adenocarcinoma that can be used in clinical practice.Result:1.SDS-PAGE: the differential expression of urine protein in the lung adenocarcinoma group and the healthy control group mainly appeared in the 90 kDa,60kDa and 20-30 kDa gel bands,while the high-abundance protein was concentrated in the 75 kDa bands.2.MS: 185 up-regulated proteins and 180 down-regulated proteins.There are 10 pro teins related to coagulation and immune pathway,which are A2 M,C9,C3,MASP2,CFB,PLG,FGG,FGB,FGA and F2.They are up-regulated proteins.3.ROC of MS data: only the sensitivity and specificity of PLG and FGG were 80%of >,and AUC>0.8.4.ELISA:(1)The expression trend of FGG and PLG was consistent with that of MS.(2)FGG and PLG had no correlation with clinical data(age,gender,smoking,clinical stage).There was no correlation between FGG and PLG.(3)ROC:The efficacy of FGG combined with PLG in differentiating lung adenocarcinoma from healthy people was significantly higher than that of single index,and the difference was statistically significant.Compared with FGG,P=0.04;compared with PLG,P=0.0008.Conclusion:Urine FGG and PLG have the potential to be diagnostic biomarkers for lung adenocarcinoma,and FGG and PLG combined mode is superior to single marker in the diagnosis of lung adenocarcinoma.