Glycoconjugate Journal

, Volume 36, Issue 1, pp 79–90 | Cite as

Identification of 9-O-acetyl-N-acetylneuraminic acid (Neu5,9Ac2) as main O-acetylated sialic acid species of GD2 in breast cancer cells

  • Sumeyye Cavdarli
  • Justine H. Dewald
  • Nao Yamakawa
  • Yann Guérardel
  • Mickaël Terme
  • Jean-Marc Le Doussal
  • Philippe Delannoy
  • Sophie Groux-DegrooteEmail author
Original Article


Mainly restricted to the nervous system in healthy adults, complex gangliosides such as GD3 and GD2 have been shown to be involved in aggressiveness and metastasis of neuro-ectoderm derived tumors such as melanoma and neuroblastoma. Interestingly, O-acetylated forms of GD2, not expressed in human peripheral nerve fibers, are highly expressed in GD2+ tumor cells. Very little information is known regarding the expression of O-acetylated disialogangliosides in breast cancer (BC) cell lines. Here, we analyzed the expression of GD2, GD3 and their O-acetylated forms O-acetyl-GD2 (OAcGD2) and O-acetyl-GD3 (OAcGD3) in BC cells. We used Hs 578T and SUM159PT cell lines, as well as cell clones over-expressing GD3 synthase derived from MDA-MB-231 and MCF-7. Using flow cytometry and immunocytochemistry/confocal microscopy, we report that BC cells express b-series gangliosides GD3 and GD2, as well as significant amounts of OAcGD2. However, OAcGD3 expression was not detected in these cells. O-acetylation of gangliosides isolated from BC cells was examined by LC-MS analysis of sialic acid DMB-derivatives. We report that the main acetylated form of sialic acid expressed in BC gangliosides is 9-O-acetyl-N-acetylneuraminic acid (Neu5,9Ac2). These results highlight a close interrelationship between Neu5,9Ac2 and OAcGD2 expression, and suggest that OAcGD2 is synthetized from GD2 and not from OAcGD3 in BC cells.


Breast Cancer Gangliosides Antibody GD2 O-acetyl-GD2 Sialic acid 



Breast Cancer


Bovine Submaxillary Mucin

CMP Neu5Ac

cytidine-monophosphate N-acetylneuraminic acid




Estrogen Receptor






GD2 synthase


GD3 synthase




Hypoxanthine-guanine PhosphoRibosylTransferase




Liquid Chromatography/ Electrospray ionization coupled to Mass Spectrometry


monoclonal antibody


Mean Fluorescence Intensity


N-acetylneuraminic acid


8-O-acetyl-N-acetylneuraminic acid


9-O-acetyl-N-acetylneuraminic acid






Tumor-Associated Carbohydrate Antigens


Triple Negative Breast Cancer



We thank Christian Slomianny of the BICeL-Campus Lille1 (Univ. Lille, Bio Imaging Center Lille, F-59000 Lille, France) facility for access to instruments and technical advices. We are indebted to the PAGés plateform (Plateforme d’Analyses des Glycoconjugués, CNRS, UMR 8576, UGSF, Université de Lille), F-59000 Lille, France for the use of the mass spectrometer.

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Sumeyye Cavdarli
    • 1
    • 2
  • Justine H. Dewald
    • 1
  • Nao Yamakawa
    • 1
  • Yann Guérardel
    • 1
  • Mickaël Terme
    • 2
  • Jean-Marc Le Doussal
    • 2
  • Philippe Delannoy
    • 1
  • Sophie Groux-Degroote
    • 1
    Email author
  1. 1.CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et FonctionnelleUniversity LilleLilleFrance
  2. 2.OGD2 PharmaInstitut de Recherche en Santé de l’Université de NantesNantesFrance

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