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.