Application of iTRAQ technique in screening bile biomarkers of biliary tract carcinoma
10.3781/j.issn.1000-7431.2024.2401-0058
- VernacularTitle:iTRAQ技术在筛选胆道肿瘤胆汁标志物中的应用
- Author:
Xiao ZHENG
1
;
Bin YAN
1
;
Tiemei LIU
1
Author Information
1. 复旦大学附属中山医院青浦分院内镜科,上海 201700
- Publication Type:Journal Article
- Keywords:
Biliary tract cancer;
Haptoglobin;
Complement Factor H;
Proteomics;
iTRAQ;
2D LC-MS/MS
- From:
Tumor
2024;44(10):993-1002
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore potential biomarkers of biliary tract carcinoma(BTC)by proteomic techniques based on isobaric tags for relative and absolute quantitation(iTRAQ).Methods:Bile samples from 4 patients with BTC and 4 patients with benign biliary tract disease(BBD)were randomly selected from sample library of central laboratory,Qingpu branch of Zhongshan Hospital,Fudan University.After mixing samples within each group,iTRAQ labeling technology combined with two-dimensional liquid chromatography-tandem mass spectrometry(2D-LC-MS/MS)was used for quantitative proteomic analysis.Then,differentially expressed proteins between the BTC and BBD groups were screened and subjected to bioinformatic analysis,including KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway enrichment analysis,GO(Gene Ontology)enrichment analysis and protein-protein interaction(PPI)network analysis.The candidate protein with the largest fold change in expression abundance selected from the differentially expressed proteins as a potential diagnostic biomarker for BTC.Enzyme-linked immunosorbent assay(ELISA)validation and receiver operating characteristic(ROC)cure analysis were performed on this candidate protein using bile samples form 50 BTC patients and 50 BBD patients.Results:A total of 12 differential expressed proteins were screened from the bile samples of 4 BTC patients and 4 BBD patients(FDR<1%).Haptoglobin(Hp)and Complement Factor H(CFH),as hub proteins in the PPI network,were significantly upregulated in the bile of BBD patients.ELISA results showed that Hp and CFH were significantly upregulated in the bile of patients with various subtypes of BTC(P<0.01).ROC curve analysis indicated that Hp and CFH in bile exhibited significant specificity and sensitivity in the diagnosis of BTC,and their diagnostic performance was superior to that of serum protein CA19-9.Conclusion:In this study,iTRAQ labeling technology combined with 2D-LC-MS/MS was used to systematically analyze the proteomic differences between bile samples from BTC and BBD patients.Hp and CFH were identified as core differentially expressed proteins,and their diagnostic efficiency for BTC was significantly better than that of serum potein CA19-9.They can serve as potential biomarkers for early diagnosis and therapeutic targets of BTC.
