Glycoproteomics Approach for Identifying Glycobiomarker Candidate Molecules for Tissue Type Classification of Non-small Cell Lung Carcinoma

Abstract

Histopathological classification of lung cancer has important implications in the application of clinical practice guidelines and the prediction of patient prognosis. Thus, we focused on discovering glycobiomarker candidates to classify the types of lung cancer tissue. First, we performed lectin microarray analysis of lung cancer tissue specimens and cell lines and identified Aleuria aurantia lectin (AAL), Hippeastrum hybrid lectin (HHL), and Concanavalia ensiformis agglutinin (ConA) as lectin probes specific to non-small cell lung carcinoma (NSCLC). LC-MS-based analysis was performed for the comprehensive identification of glycoproteins and N-linked glycosylation sites using lectin affinity capture of NSCLC-specific glycoforms of glycoproteins. This analysis identified 1092 AAL-bound glycoproteins (316 gene symbols) and 948 HHL/ConA-bound glycoproteins (279 gene symbols). The lectin microarray-assisted verification using 15 lung cancer cell lines revealed the NSCLC-specific expression of fibronectin. The glycosylation profiling of fibronectin indicated that the peanut agglutinin (PNA) signal appeared to differentiate two NSCLC types, adenocarcinoma and large cell carcinoma, whereas the protein expression level was similar between these types. Our glycoproteomics approach together with the concurrent use of an antibody and lectin is applicable to the quantitative and qualitative monitoring of variations in glycosylation of fibronectin specific to certain types of lung cancer tissue.

Identification and characterization of sulfated glycoproteins from small cell lung carcinoma cells assisted by management of molecular charges

“…The glycan structures on these proteins, however, have not been elucidated. The neuroendocrine marker NCAM1 was detected only in SCLC cell lines [42]. Upregulation of APLP1 has been reported in neuroendocrine tumors [43].…”

“…Thus, these sulfated glycoproteins may be relatively abundant in the serum-free culture medium of H2171 cells. Recently, Hirao et al identified GOLM1 and IGFBPL1 in the culture media from SCLC and NSCLC cells using lectin-based glycoproteomic analysis [42]. The glycan structures on these proteins, however, have not been elucidated.…”

Plant Lectins Targeting O-Glycans at the Cell Surface as Tools for Cancer Diagnosis, Prognosis and Therapy

“…A lectin-based microarray analysis discriminated between healthy patients and patients with colorectal cancer due to the higher degree a α2,6-sialylation and the higher content in high mannose N -glycans in serum α2-macroglobulin [125]. Glycosylation profiling of fibronectin performed with a lectin-microarray, showed that PNA distinguishes between two distinct types of non-small cell lung carcinomas, lung adenocarcinoma and large cell lung carcinoma [126]. In a lectin-microarray analysis including the T/Tn-specific lectins ACA ( Amaranthus caudatus ), ACG ( Agrocybe cylindracea ), BPL ( Bauhinia purpurea ), and SNA ( Sambucus nigra ), a decreased lectin-binding activity was observed for Tn antigen from formalin-fixed human choriocarcinoma tissues [24].…”

Development of M2BPGi: a novel fibrosis serum glyco-biomarker for chronic hepatitis/cirrhosis diagnostics

“…For instance, lung cancer can be classified into small cell carcinoma, adenocarcinoma, squamous cell carcinoma and large cell carcinoma, while ovarian cancer is classified into clear cell carcinoma, mucinous adenocarcinoma, serous adenocarcinoma and endometrioid adenocarcinoma. If the histological types of cancer in the same organ are different, glycan structures and probing lectins are largely different and distinctive [14,46]. Therefore, a collaborative study with experienced clinical doctors and pathologists is essential.…”

“…For instance, structures of glycans are different between small cell lung carcinoma and non-small-cell lung carcinoma [14], or among hepatoma, hepatocellular carcinoma and intrahepatic bile duct carcinoma [15]. Glycans are also expected to be useful as specific markers in the research and production of stem cells and induced pluripotent stem cells [16].…”