Urine Differential Protein Analysis And Early Diagnosis Model Of Bladder Urothelial Carcinoma Based On R Language

Objectives To find the differential protein of BUC urine and verify,a data set of BUC urine differentially expressed proteins was established for early diagnosis and discrimination model of BUC.Methods By using of Isobaric tags for relative and absolute quantitation(iTRAQ),a total of 6 urine specimens,including 4 cases in BUC group and 2 cases in control group,were with the intention of comparative proteomics research,so as to find out the differentially expressed proteins in urine between the two groups.The top 20 of log2 fold change differentially expressed proteins were selected to perform MRM verification;KEGG and GO enrichment analysis by using enrichplot and org.Hs.eg.db package respectively.Significant differential proteins were preliminarily selected to construct the BUC urine differential protein data set.The STRING website was used to draw the Protein Protein Interaction(PPI)diagram to screen the differentially expressed proteins.The urine expression level of the core differential protein selected according to PPI was estimated by ELISA,the Receiver operator characteristic(ROC)curve was applied to determine the optimal working point of each differential protein and the combination of the differential protein for the early diagnosis of BUC,and the corresponding specificity and sensitivity were calculated.In addition,anther diagnosis model based on a logistic model was established to fit the differential protein dose estimated using ELISA both in the BUC group and the control group.Results 1 8823 peptide and 1949 protein were detected by iTRAQ.101 BUC urine differential proteins were obtained including 37 proteins up-regulated and 64 proteins were down-regulated.2 MRM confirmed the existence of 10 differential proteins,A2 MG,DESP,APOE,TPM3,TPOA4,OTUBI,BPI,PCD19,RS14 and POK18,respectively.3The KEGG enrichment analysis showed significantly different protein concentration in other chitosan degradation,complement and blood coagulation cascade both metabolic pathways,containing 11 differentially expressed proteins.4 GO analysis showed that differential proteins were mainly involved in cell adhesion molecule binding,carboxylic acid binding,organic acid binding,glycosaminoglycan binding,peptide-chain enzyme activation and other biological processes,54 differential proteins were involved.5 The three together constitute the BUC urine differential protein data set,including 69 proteins.6 By protein interaction analysis,APOE and APOA4 were selected as the hub differential proteins.7 The dose of APOE and APOA4 in urine sample from BUC patients and healthy population were estimated using ELISA.8 A ROC plot was drawn using these data.For APOE,the area under the curve(AUC)is 0.941,the optimal operating point(OOP)is0.41(pg/ml),the sensitivity is 0.875 and the specificity is 0.889;For APOA4,the area under the curve(AUC)is 0.904,the optimal operating point(OOP)is 4.51(ng/ml),the sensitivity is 0.875 and the specificity is 0.919.9.The regression coefficients are 18.837 and 1.077 for APOE and APOA4 respectively.The results are statistically significant(P<0.01).Conclusions 1.BUC urine differential protein data set including 69 differential protein was constructed.2.BUC differential proteins were mainly concentration on other chitosan degradation,complement and blood coagulation cascade both metabolic pathways,and involved in cell adhesion molecule binding,carboxylic acid binding,organic acid binding,glycosaminoglycan binding,peptide-chain enzyme activation and other biological processes.3.BUC urine differential protein APOE and APOA4 have the potential to be used in BUC diagnosis.Figure9,Table8,Reference 71...