Depletion of β1,6-N-acetylglucosaminyltransferase reduces E-selectin binding capacity and migratory potential of human gastrointestinal adenocarcinoma cells

The commonly altered glycosylation of tumor cells is a hallmark of tumor progression and metastasis formation. One prominent example is the interaction of sialylated glycans at the tumor cell surface with endothelial (E)-selectin as an early event of an adhesion cascade that enables extravasation of...

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Main Authors: Lisa Staffeldt, Hanna Maar, Julia Beimdiek, Samuel Chambers, Kristoffer Riecken, Mark von Itzstein, Falk F.R. Buettner, Arun Everest-Dass, Tobias Lange
Format: Article
Language:English
Published: Elsevier 2025-01-01
Series:Neoplasia: An International Journal for Oncology Research
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Online Access:http://www.sciencedirect.com/science/article/pii/S1476558624001246
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author Lisa Staffeldt
Hanna Maar
Julia Beimdiek
Samuel Chambers
Kristoffer Riecken
Mark von Itzstein
Falk F.R. Buettner
Arun Everest-Dass
Tobias Lange
author_facet Lisa Staffeldt
Hanna Maar
Julia Beimdiek
Samuel Chambers
Kristoffer Riecken
Mark von Itzstein
Falk F.R. Buettner
Arun Everest-Dass
Tobias Lange
author_sort Lisa Staffeldt
collection DOAJ
description The commonly altered glycosylation of tumor cells is a hallmark of tumor progression and metastasis formation. One prominent example is the interaction of sialylated glycans at the tumor cell surface with endothelial (E)-selectin as an early event of an adhesion cascade that enables extravasation of circulating tumor cells (CTCs) into distant tissues. In a previous study, we identified GCNT3 (mucin-type core2/ core4 β1,6-N-acetylglucosaminyltransferase) highly over-expressed in gastrointestinal adenocarcinoma cells that facilitate the canonical E-selectin ligands sialyl-Lewis A and X (sLeA/X) for E-selectin binding and endothelial adhesion. Here we show that shRNA-mediated, stable depletion of GCNT3 reduced sLeA (tumor marker CA19-9) presentation on two out of three tested human gastrointestinal adenocarcinoma cell lines, concurrently showing reduced static E-selectin binding. Significant effects of GCNT3 depletion on dynamic, shear-resistant tumor cell adhesion on immobilized E-selectin as well as endothelial cells were only partially and inconsistently observable as were effects on tumor cell proliferation (2D) or 3D colony formation. Nevertheless, tumor cell migration was consistently reduced upon GCNT3 depletion in all tested cell lines. Detailed glycome analyses revealed that GCNT3 depletion caused cell line-specific alterations in N- and O-glycans as well as glycosphingolipids, collectively mainly associating with decreased Core-2 structures resulting in varied abundance of sialylation and Lewis antigen with consistent phenotypic changes. Distinctive N- and O-glycosylation features were found to be inherent to specific cell types. These findings suggest GCNT3 products as possible carriers of sLeA and static E-selectin binding sites as well as common pro-migratory glycans in human gastrointestinal cancer.
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spelling doaj-art-ef40da9ca4364521a9d375bdd0324fd32024-12-17T04:59:01ZengElsevierNeoplasia: An International Journal for Oncology Research1476-55862025-01-0159101083Depletion of β1,6-N-acetylglucosaminyltransferase reduces E-selectin binding capacity and migratory potential of human gastrointestinal adenocarcinoma cellsLisa Staffeldt0Hanna Maar1Julia Beimdiek2Samuel Chambers3Kristoffer Riecken4Mark von Itzstein5Falk F.R. Buettner6Arun Everest-Dass7Tobias Lange8Institute of Anatomy and Experimental Morphology, University Cancer Center Hamburg, University Medical Center Hamburg-Eppendorf, 20241, Hamburg, GermanyInstitute of Anatomy and Experimental Morphology, University Cancer Center Hamburg, University Medical Center Hamburg-Eppendorf, 20241, Hamburg, Germany; Institute of Anatomy I, Jena University Hospital, 07743, Jena, Germany; Comprehensive Cancer Center Central Germany (CCCG)Institute of Clinical Biochemistry, Hannover Medical School, 30625, Hannover, Germany; Proteomics, Institute of Theoretical Medicine, Faculty of Medicine, University of Augsburg, Augsburg, GermanyInstitute for Glycomics, Griffith University, Gold Coast Campus, Gold Coast, QLD4222, AustraliaResearch Department Cell and Gene Therapy, Department of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf, 20246, HamburgInstitute for Glycomics, Griffith University, Gold Coast Campus, Gold Coast, QLD4222, AustraliaInstitute of Anatomy and Experimental Morphology, University Cancer Center Hamburg, University Medical Center Hamburg-Eppendorf, 20241, Hamburg, Germany; Institute of Clinical Biochemistry, Hannover Medical School, 30625, Hannover, Germany; Proteomics, Institute of Theoretical Medicine, Faculty of Medicine, University of Augsburg, Augsburg, GermanyInstitute of Anatomy and Experimental Morphology, University Cancer Center Hamburg, University Medical Center Hamburg-Eppendorf, 20241, Hamburg, Germany; Institute for Glycomics, Griffith University, Gold Coast Campus, Gold Coast, QLD4222, AustraliaInstitute of Anatomy and Experimental Morphology, University Cancer Center Hamburg, University Medical Center Hamburg-Eppendorf, 20241, Hamburg, Germany; Institute of Anatomy I, Jena University Hospital, 07743, Jena, Germany; Comprehensive Cancer Center Central Germany (CCCG); Corresponding author.The commonly altered glycosylation of tumor cells is a hallmark of tumor progression and metastasis formation. One prominent example is the interaction of sialylated glycans at the tumor cell surface with endothelial (E)-selectin as an early event of an adhesion cascade that enables extravasation of circulating tumor cells (CTCs) into distant tissues. In a previous study, we identified GCNT3 (mucin-type core2/ core4 β1,6-N-acetylglucosaminyltransferase) highly over-expressed in gastrointestinal adenocarcinoma cells that facilitate the canonical E-selectin ligands sialyl-Lewis A and X (sLeA/X) for E-selectin binding and endothelial adhesion. Here we show that shRNA-mediated, stable depletion of GCNT3 reduced sLeA (tumor marker CA19-9) presentation on two out of three tested human gastrointestinal adenocarcinoma cell lines, concurrently showing reduced static E-selectin binding. Significant effects of GCNT3 depletion on dynamic, shear-resistant tumor cell adhesion on immobilized E-selectin as well as endothelial cells were only partially and inconsistently observable as were effects on tumor cell proliferation (2D) or 3D colony formation. Nevertheless, tumor cell migration was consistently reduced upon GCNT3 depletion in all tested cell lines. Detailed glycome analyses revealed that GCNT3 depletion caused cell line-specific alterations in N- and O-glycans as well as glycosphingolipids, collectively mainly associating with decreased Core-2 structures resulting in varied abundance of sialylation and Lewis antigen with consistent phenotypic changes. Distinctive N- and O-glycosylation features were found to be inherent to specific cell types. These findings suggest GCNT3 products as possible carriers of sLeA and static E-selectin binding sites as well as common pro-migratory glycans in human gastrointestinal cancer.http://www.sciencedirect.com/science/article/pii/S1476558624001246β1,6-N-acetylglucosaminyltransferaseGCNT3Gastrointestinal adenocarcinomaE-selectinSialyl-Lewis ATumor cell migration
spellingShingle Lisa Staffeldt
Hanna Maar
Julia Beimdiek
Samuel Chambers
Kristoffer Riecken
Mark von Itzstein
Falk F.R. Buettner
Arun Everest-Dass
Tobias Lange
Depletion of β1,6-N-acetylglucosaminyltransferase reduces E-selectin binding capacity and migratory potential of human gastrointestinal adenocarcinoma cells
Neoplasia: An International Journal for Oncology Research
β1,6-N-acetylglucosaminyltransferase
GCNT3
Gastrointestinal adenocarcinoma
E-selectin
Sialyl-Lewis A
Tumor cell migration
title Depletion of β1,6-N-acetylglucosaminyltransferase reduces E-selectin binding capacity and migratory potential of human gastrointestinal adenocarcinoma cells
title_full Depletion of β1,6-N-acetylglucosaminyltransferase reduces E-selectin binding capacity and migratory potential of human gastrointestinal adenocarcinoma cells
title_fullStr Depletion of β1,6-N-acetylglucosaminyltransferase reduces E-selectin binding capacity and migratory potential of human gastrointestinal adenocarcinoma cells
title_full_unstemmed Depletion of β1,6-N-acetylglucosaminyltransferase reduces E-selectin binding capacity and migratory potential of human gastrointestinal adenocarcinoma cells
title_short Depletion of β1,6-N-acetylglucosaminyltransferase reduces E-selectin binding capacity and migratory potential of human gastrointestinal adenocarcinoma cells
title_sort depletion of β1 6 n acetylglucosaminyltransferase reduces e selectin binding capacity and migratory potential of human gastrointestinal adenocarcinoma cells
topic β1,6-N-acetylglucosaminyltransferase
GCNT3
Gastrointestinal adenocarcinoma
E-selectin
Sialyl-Lewis A
Tumor cell migration
url http://www.sciencedirect.com/science/article/pii/S1476558624001246
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