Probing the O-Glycoproteome of Gastric Cancer Cell Lines for Biomarker Discovery*
| Autor: | Sergey Y. Vakhrushev, Daniela Freitas, J. Sousa, Catharina Steentoft, Luís Pedro Afonso, Catarina Gomes, Diana Campos, Ana Magalhães, Celso A. Reis, Malene Bech Vester-Christensen, Ulla Mandel, Lúcio Lara Santos, Henrik Clausen, Joana Gomes, José Alexandre Ferreira |
|---|---|
| Přispěvatelé: | Instituto de Investigação e Inovação em Saúde |
| Rok vydání: | 2015 |
| Předmět: |
Male
N-Acetylgalactosaminyltransferases/blood Proteome Glycoproteins/metabolism Proximity ligation assay Biology Stomach Neoplasms/metabolism Biochemistry Analytical Chemistry 03 medical and health sciences 0302 clinical medicine Stomach Neoplasms Cell Line Tumor Biomarkers Tumor medicine N-Acetylgalactosaminyltransferases/metabolism Humans Biomarker discovery Glycoproteins/blood Molecular Biology Aged Glycoproteins 030304 developmental biology Aged 80 and over chemistry.chemical_classification 0303 health sciences Research CD44 Cancer Middle Aged medicine.disease 3. Good health chemistry Biomarkers Tumor/blood Biomarkers Tumor/metabolism 030220 oncology & carcinogenesis Cancer cell Immunology Cancer research biology.protein Stomach Neoplasms/blood N-Acetylgalactosaminyltransferases Biomarker (medicine) Female Glycoprotein |
| Zdroj: | Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP |
| ISSN: | 1535-9476 |
| DOI: | 10.1074/mcp.m114.046862 |
| Popis: | Circulating O-glycoproteins shed from cancer cells represent important serum biomarkers for diagnostic and prognostic purposes. We have recently shown that selective detection of cancer-associated aberrant glycoforms of circulating O-glycoprotein biomarkers can increase specificity of cancer biomarker assays. However, the current knowledge of secreted and circulating O-glycoproteins is limited. Here, we used the COSMC KO "Simple- Cell" (SC) strategy to characterize the O-glycoproteome of two gastric cancer SimpleCell lines (AGS, MKN45) as well as a gastric cell line (KATO III) which naturally expresses at least partially truncated O-glycans. Overall, we identified 499 O-glycoproteins and 1236 O-glycosites in gastric cancer SimpleCells, and a total 47 O-glycoproteins and 73 O-glycosites in the KATO III cell line. We next modified the glycoproteomic strategy to apply it to pools of sera from gastric cancer and healthy individuals to identify circulating O-glycoproteins with the STn glycoform. We identified 37 O-glycoproteins in the pool of cancer sera, and only nine of these were also found in sera from healthy individuals. Two identified candidate O-glycoprotein biomarkers (CD44 and GalNAc-T5) circulating with the STn glycoform were further validated as being expressed in gastric cancer tissue. A proximity ligation assay was used to show that CD44 was expressed with the STn glycoform in gastric cancer tissues. The study provides a discovery strategy for aberrantly glycosylated O-glycoproteins and a set of O-glycoprotein candidates with biomarker potential in gastric cancer. This work was supported by The Danish Research Councils, The Mizutani Foundation, The Danish National Research Foundation (DNRF107) and Fundacão para a Ciência e a Tecnologia (FCT) and COMPETE (Programa Operacional Temático Factores de Competitividade, comparticipado pelo fundo comunitário europeu FEDER) in the framework of the projects: PTDC/BBB-EBI/0786/2012; EXPL/CTM-BIO/0762/2013. Grants were received from FCT (SFRH/BD/73717/2010 to DC), (SFRH/BPD/75871/2011 to AM), (SFRH/BPD/96510/2013 to CG) and (SFRH/BPD/66288/2009 to JAF). IPATIMUP is an Associate Laboratory of the Portuguese Ministry of Science, Technology and Higher Education, and is partially supported by FCT. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|