Novel Glycobiomarker for Ovarian Cancer That Detects Clear Cell Carcinoma

• Published in: Journal of proteome research Vol. 13; no. 3; pp. 1624 - 1635
• Main Authors: Sogabe, Maki, Nozaki, Hirofumi, Tanaka, Nana, Kubota, Tomomi, Kaji, Hiroyuki, Kuno, Atsushi, Togayachi, Akira, Gotoh, Masanori, Nakanishi, Hayao, Nakanishi, Toru, Mikami, Mikio, Suzuki, Nao, Kiguchi, Kazushige, Ikehara, Yuzuru, Narimatsu, Hisashi
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 07.03.2014
• Subjects: Adenocarcinoma, Clear Cell - chemistry; Adenocarcinoma, Clear Cell - diagnosis; Ascitic Fluid - chemistry; Biomarkers, Tumor - analysis; CA-125 Antigen - analysis; Carcinoma, Ovarian Epithelial; Ceruloplasmin - analysis; Ceruloplasmin - chemistry; Chromatography, Liquid; Female; Glycoproteins - analysis; Humans; Mass Spectrometry; Neoplasms, Glandular and Epithelial - chemistry; Neoplasms, Glandular and Epithelial - diagnosis; Ovarian Neoplasms - chemistry; Ovarian Neoplasms - diagnosis; Plant Lectins - chemistry; Polysaccharides - analysis; Polysaccharides - chemistry; Protein Array Analysis; Receptors, N-Acetylglucosamine - chemistry; glycan alteration; glycoproteomics; ceruloplasmin; WFA; clear cell carcinoma; lectin microarray; glycoprotein; epithelial ovarian cancer
• ISSN: 1535-3893; 1535-3907
• DOI: 10.1021/pr401109n

Epithelial ovarian cancer (EOC) is often asymptomatic and thus diagnosed at advanced stages with a poor prognosis. False-negative results for the conventional marker CA125 frequently occur in cases of clear cell carcinoma (CCC), a type of EOC; therefore, it is necessary to develop biomarkers with greater sensitivity. We previously reported a strategy to discover glycobiomarker candidates by combined lectin microarray and IGOT–LC/MS analysis. We have now optimized this strategy for discovering EOC biomarkers. Glycopeptides possessing cancerous glycans were enriched from the ascites fluids and culture supernatants of cancer cell lines with a fucose-binding lectin, AAL. IGOT–LC/MS analysis of CCC samples yielded 144 candidate glycoproteins. We selected WFA by lectin microarray as the optimal lectin to distinguish EOC from gastric and colon cancer. The candidates were narrowed by Western analysis of the WFA-bound fraction of ascites fluids. One of the final candidates, WFA-reactive ceruloplasmin, produced higher signals in the ascites fluids of EOC patients, including CCC, in comparison with the benign samples, while CA125 levels were comparable in the sandwich ELISA. Thus, our glycoproteomic strategy featuring efficient enrichment of glycans with disease-related alterations is applicable to various diseases.