title: Identification of GlcNAcylated alpha-1-antichymotrypsin as an early biomarker in human non-small-cell lung cancer by quantitative proteomic analysis with two lectins.
abstract: BACKGROUND
  Non-small-cell lung cancer (NSCLC) is the main type of lung cancer with high mortality rates in worldwide. There is a need to identify better biomarkers to detect NSCLC at an early stage as this will improve therapeutic effect and patient survival rates.
  
  
  METHODS
  Two lectins (AAL/AAGL and AAL2/AANL), which specifically bind to tumour-related glycan antigens, were first used to enrich serum glycoproteins from the serum of early NSCLC patients, benign lung diseases subjects and healthy individuals. The samples were investigated by using iTRAQ labelling and LC-MS/MS.
  
  
  RESULTS
  A total of 53 differentially expressed proteins were identified by quantitative proteomics and four glycoproteins (AACT, AGP1, CFB and HPX) were selected for further verification by western blotting. Receiver operating characteristic analysis showed AACT was the best candidate for early NSCLC diagnosis of the four proteins, with 94.1% sensitivity in distinguishing early tumour Stage (IA+IB) from tumour-free samples (healthy and benign samples, HB). The GlcNAcylated AACT was further detected by lectin-based ELISA and has better advantage in clinical application than total AACT. The GlcNAcylated AACT can effectively differentiate Stage I from HB samples with an AUC of 0.908 and 90.9% sensitivity at a specificity of 86.2%. A combination of GlcNAcylated AACT and carcinoembryonic antigen (CEA) was able to effectively differing Stage I from HB samples (AUC=0.914), which significantly improve the specificity of CEA. The combination application also has the better clinical diagnostic efficacy in distinguishing cancer (NSCLC) from HB samples than CEA or GlcNAcylated AACT used alone, and yielded an AUC of 0.817 with 93.1% specificity.
  
  
  CONCLUSIONS
  Our findings suggest that the GlcNAcylated AACT will be a promising clinical biomarker in diagnosis of early NSCLC.
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Yanxia
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Department of Thoracic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, P.R. China.
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Ye
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Yanting
authors.initials: Y
authors.lastname: Su
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Guojun
authors.initials: G
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Wenhui
authors.initials: W
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Jie
authors.initials: J
authors.lastname: Cai
authors.affiliation: Wuhan Institute of Biotechnology, Wuhan 430075, Hubei Province, P.R. China.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Clinical Immunology, Guangdong Medical College, Dongguan 523808, Hubei Province, P.R. China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Yijie
authors.initials: Y
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Barry Hon Cheung
authors.initials: BH
authors.lastname: Wong
authors.affiliation: Department of Thoracic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, P.R. China.
authors.forename: Xiangning
authors.initials: X
authors.lastname: Fu
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, P.R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Sun
mesh_terms: D000328:Adult; D000368:Aged; D042683:Angiopoietin-1 / Q000378:metabolism*; D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism*; D015153:Blotting, Western; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis / Q000378:metabolism*; D016022:Case-Control Studies; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D011415:Complement Factor B / Q000378:metabolism*; D004195:Disease Models, Animal; D005260:Female; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D006466:Hemopexin / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D037102:Lectins; D008175:Lung Neoplasms / Q000175:diagnosis / Q000378:metabolism*; D008297:Male; D051379:Mice; D008875:Middle Aged; D040901:Proteomics; D012372:ROC Curve; D015843:Serpins / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D055815:Young Adult
chemical_list: C467630:ANGPT1 protein, human; D042683:Angiopoietin-1; D014408:Biomarkers, Tumor; D006023:Glycoproteins; D037102:Lectins; C516148:SERPINA3 protein, human; D015843:Serpins; D006466:Hemopexin; D011415:Complement Factor B