Glycomic analysis of gastric carcinoma cells discloses glycans as modulators of RON receptor tyrosine kinase activation in cancer

Stefan Mereiter; Ana Magalhães; Barbara Adamczyk; Chunsheng Jin; Andreia Almeida; Lylia Drici; Maria Ibáñez-Vea; Catarina Gomes; José A Ferreira; Luis P Afonso; Lúcio L Santos; Martin R Larsen; Daniel Kolarich; Niclas G Karlsson; Celso A Reis

doi:10.1016/j.bbagen.2015.12.016

Glycomic analysis of gastric carcinoma cells discloses glycans as modulators of RON receptor tyrosine kinase activation in cancer

Biochim Biophys Acta. 2016 Aug;1860(8):1795-808. doi: 10.1016/j.bbagen.2015.12.016. Epub 2015 Dec 22.

Authors

Affiliations

¹ I3S - Instituto de Investigação e Inovação em Saúde, University of Porto, Portugal; Institute of Molecular Pathology and Immunology of the University of Porto - IPATIMUP, Porto, Portugal; Institute of Biomedical Sciences of Abel Salazar - ICBAS, University of Porto, Portugal.
² I3S - Instituto de Investigação e Inovação em Saúde, University of Porto, Portugal; Institute of Molecular Pathology and Immunology of the University of Porto - IPATIMUP, Porto, Portugal.
³ Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Sweden.
⁴ Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, 14424 Potsdam, Germany; Free University Berlin, Berlin, Germany.
⁵ Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
⁶ I3S - Instituto de Investigação e Inovação em Saúde, University of Porto, Portugal; Institute of Molecular Pathology and Immunology of the University of Porto - IPATIMUP, Porto, Portugal; Experimental Pathology and Therapeutics Group, Portuguese Institute of Oncology of Porto, Portugal.
⁷ Department of Pathology, Portuguese Institute of Oncology of Porto, Portugal.
⁸ Experimental Pathology and Therapeutics Group, Portuguese Institute of Oncology of Porto, Portugal; Department of Surgical Oncology, Portuguese Institute of Oncology of Porto, Portugal.
⁹ Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, 14424 Potsdam, Germany.
¹⁰ I3S - Instituto de Investigação e Inovação em Saúde, University of Porto, Portugal; Institute of Molecular Pathology and Immunology of the University of Porto - IPATIMUP, Porto, Portugal; Institute of Biomedical Sciences of Abel Salazar - ICBAS, University of Porto, Portugal; Medical Faculty, University of Porto, Portugal. Electronic address: celsor@ipatimup.pt.

PMID: 26721331
DOI: 10.1016/j.bbagen.2015.12.016

Abstract

Background: Terminal α2-3 and α2-6 sialylation of glycans precludes further chain elongation, leading to the biosynthesis of cancer relevant epitopes such as sialyl-Lewis X (SLe(X)). SLe(X) overexpression is associated with tumor aggressive phenotype and patients' poor prognosis.

Methods: MKN45 gastric carcinoma cells transfected with the sialyltransferase ST3GAL4 were established as a model overexpressing sialylated terminal glycans. We have evaluated at the structural level the glycome and the sialoproteome of this gastric cancer cell line applying liquid chromatography and mass spectrometry. We further validated an identified target expression by proximity ligation assay in gastric tumors.

Results: Our results showed that ST3GAL4 overexpression leads to several glycosylation alterations, including reduced O-glycan extension and decreased bisected and increased branched N-glycans. A shift from α2-6 towards α2-3 linked sialylated N-glycans was also observed. Sialoproteomic analysis further identified 47 proteins with significantly increased sialylated N-glycans. These included integrins, insulin receptor, carcinoembryonic antigens and RON receptor tyrosine kinase, which are proteins known to be key players in malignancy. Further analysis of RON confirmed its modification with SLe(X) and the concomitant activation. SLe(X) and RON co-expression was validated in gastric tumors.

Conclusion: The overexpression of ST3GAL4 interferes with the overall glycophenotype of cancer cells affecting a multitude of key proteins involved in malignancy. Aberrant glycosylation of the RON receptor was shown as an alternative mechanism of oncogenic activation.

General significance: This study provides novel targets and points to an integrative tumor glycomic/proteomic-profiling for gastric cancer patients' stratification. This article is part of a Special Issue entitled "Glycans in personalised medicine" Guest Editor: Professor Gordan Lauc.

Keywords: Gastric cancer; Glycome; RON; ST3GAL4; Sialome; Sialyl Lewis X (SLeX).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Gene Expression Regulation, Enzymologic*
Gene Expression Regulation, Neoplastic*
Glycomics
Humans
Lewis X Antigen / biosynthesis*
Lewis X Antigen / genetics
Neoplasm Proteins / biosynthesis*
Neoplasm Proteins / genetics
Polysaccharides / biosynthesis*
Polysaccharides / genetics
Receptor Protein-Tyrosine Kinases / genetics
Receptor Protein-Tyrosine Kinases / metabolism*
Sialyl Lewis X Antigen
Sialyltransferases / biosynthesis
Sialyltransferases / genetics
Stomach Neoplasms / genetics
Stomach Neoplasms / metabolism*
beta-Galactoside alpha-2,3-Sialyltransferase

Substances

Lewis X Antigen
Neoplasm Proteins
Polysaccharides
Sialyl Lewis X Antigen
Sialyltransferases
RON protein
Receptor Protein-Tyrosine Kinases
beta-Galactoside alpha-2,3-Sialyltransferase
ST3GAL1 protein, human