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Molecular Cancer
Published in: Molecular Cancer 1/2015

Open Access 01-12-2015 | Research

COSMC knockdown mediated aberrant O-glycosylation promotes oncogenic properties in pancreatic cancer

Authors: Bianca T. Hofmann, Laura Schlüter, Philip Lange, Baris Mercanoglu, Florian Ewald, Aljonna Fölster, Aeint-Steffen Picksak, Sönke Harder, Alexander T. El Gammal, Katharina Grupp, Cenap Güngör, Astrid Drenckhan, Hartmut Schlüter, Christoph Wagener, Jakob R. Izbicki, Manfred Jücker, Maximilian Bockhorn, Gerrit Wolters-Eisfeld

Published in: Molecular Cancer | Issue 1/2015

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Abstract

Background

Human pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive and lethal malignancies in the world and despite great efforts in research types of treatment remain limited. A frequently detected alteration in PDACs is a truncated O-linked N-acetylgalactosamine (GalNAc) glycosylation with expression of the Tn antigen. Changes in O-glycosylation affect posttranslationally modified O-GalNAc proteins resulting in profound cellular alterations. Tn antigen is a tumor associated glycan detected in 75-90 % of PDACs and up to 67 % in its precursor lesions. Since the role of Tn antigen expression in PDAC is insufficiently understood we analyzed the impact of COSMC mediated Tn antigen expression in two human PDAC cell lines on cellular oncogenic properties.

Methods

Forced expression of Tn antigen on O-glycosylated proteins in pancreatic cancer cells was induced by lentiviral-mediated knockdown of the COSMC chaperone, which prevented O-glycan elongation beyond the initial GalNAcα1- residue on O-linked glycoproteins. Altered O-GalNAc glycosylation was analyzed in human pancreatic cancer cell lines Panc-1 and L3.6pl using Western and Far-Western blot as well as immunocytochemical techniques. To assess the biological implications of COSMC function on oncogenic properties, cell viability assays, scratch assays combined with live cell imaging, migration and apoptosis assays were performed. Lectin based glycoprotein enrichment with subsequent mass spectrometric analysis identified new cancer O-GalNAc modified proteins. Expression of Tn antigen bearing Nucleolin in patient derived PDAC tumor specimens was evaluated and correlated with clinicopathological data.

Results

Tn antigen expression was induced on various O-GalNAc glycoproteins in COSMC deficient cell lines compared to the control. Proliferation was reduced (p < 0.001) in COSMC knockdown cells, whereas migration was increased (p < 0.001) and apoptosis was decreased (p = 0.03), highlighting the importance of Tn antigen expression on metastatic and anti-apoptotic behavior of PDAC derived cells. Nucleolin was identified as O-GalNAc modified protein in COSMC deficient PDAC cell lines. Interestingly, immunohistochemical staining and co-localization studies of patient derived PDACs revealed poor survival for patients with strong co-localization of Tn antigen and Nucleolin (p = 0.037).

Conclusion

This study substantiates the influence of altered O-glycan (Tn/STn) expression on oncogenic properties in pancreatic cancer and identifies O-GalNAc modified Nucleolin as novel prognostic marker.
Appendix
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Metadata
Title
COSMC knockdown mediated aberrant O-glycosylation promotes oncogenic properties in pancreatic cancer
Authors
Bianca T. Hofmann
Laura Schlüter
Philip Lange
Baris Mercanoglu
Florian Ewald
Aljonna Fölster
Aeint-Steffen Picksak
Sönke Harder
Alexander T. El Gammal
Katharina Grupp
Cenap Güngör
Astrid Drenckhan
Hartmut Schlüter
Christoph Wagener
Jakob R. Izbicki
Manfred Jücker
Maximilian Bockhorn
Gerrit Wolters-Eisfeld
Publication date
01-12-2015
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2015
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/s12943-015-0386-1

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