Inhibition of human neuroblastoma cell proliferation and EGF receptor phosphorylation by gangliosides GM1, GM3, GD1A and GT1B

Cell Prolif. 2002 Apr;35(2):105-15. doi: 10.1046/j.1365-2184.2002.00228.x.


The inhibitory action of gangliosides GT1B, GD1A, GM3 and GM1 on cell proliferation and epidermal growth factor receptor (EGFR) phosphorylation was determined in the N-myc amplified human neuroblastoma cell line NBL-W. The IC50 of each ganglioside was estimated from concentration-response regressions generated by incubating NBL-W cells with incremental concentrations (5-1000 microm) of GT1B, GD1A, GM3 or GM1 for 4 days. Cell proliferation was quantitatively determined by a colourimetric assay using tetrazolium dye and spectrophotometric analysis, and EGFR phosphorylation by densitometry of Western blots. All gangliosides assayed, with the exception of GM1, inhibited NBL-W cell proliferation in a concentration-dependent manner. The IC50s for gangliosides GT1B [molecular weight (MW) 2129], GM3 (MW 1236), and GD1A (MW 1838) were (mean +/- SEM) 117 +/- 26, 255 +/- 29, and 425 +/- 44 m, respectively. In contrast, the IC50 for GM1 (MW 1547) could not be determined. Incubation of NBL-W cells with epidermal growth factor (EGF) concentrations ranging from 0.1 to 1000 ng/ml progressively increased cell proliferation rate, but it plateaued at concentrations above 10 ng/ml. EGFR tyrosine phosphorylation, however, was incrementally stimulated by EGF concentrations from 1 to 100 ng/ml. The suppression of EGF-induced EGFR phosphorylation differed for each ganglioside, and their respective inhibitory potencies were as follows: EGFR phosphorylation [area under curve (+ EGF)/area under curve (- EGF)]: control (no ganglioside added) = 8.2; GM1 = 8.3; GD1A = 6.7; GM3 = 4.87, and GT1B = 4.09. The lower the ratio, the greater the inhibitory activity of the ganglioside. Gangliosides GD1A and GT1B, which have terminal N-acetyl neuraminic acid moieties, as well as one and two N-acetyl neuraminic acid residues linked to the internal galactose, respectively, both inhibited cell proliferation and EGFR phosphorylation. However, GD1A was a more potent suppressor of cell proliferation and GT1B most effective against EGFR phosphorylation. GM3, which only has a terminal N-acetyl neuraminic acid, inhibited cell proliferation and EGFR phosphorylation almost equivalently. These data suggest that gangliosides differ in their potency as inhibitors of NBL-W neuroblastoma cell proliferation and EGFR tyrosine phosphorylation, and that perturbations in the differential expression of membrane glycosphingolipids may play a role in modulating neuroblastoma growth.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbohydrate Sequence
  • Cell Division / drug effects
  • Cell Division / physiology
  • Dogs
  • Epidermal Growth Factor / pharmacology
  • ErbB Receptors / metabolism*
  • G(M1) Ganglioside / chemistry
  • G(M1) Ganglioside / metabolism*
  • G(M1) Ganglioside / pharmacology
  • G(M3) Ganglioside / chemistry
  • G(M3) Ganglioside / metabolism*
  • G(M3) Ganglioside / pharmacology
  • Gangliosides / chemistry
  • Gangliosides / metabolism
  • Gangliosides / pharmacology
  • Humans
  • Molecular Sequence Data
  • Neuroblastoma*
  • Phosphorylation
  • Structure-Activity Relationship
  • Tumor Cells, Cultured / cytology
  • Tumor Cells, Cultured / metabolism
  • Tyrosine / metabolism


  • G(M3) Ganglioside
  • Gangliosides
  • ganglioside, GD1a
  • G(M1) Ganglioside
  • Tyrosine
  • trisialoganglioside GT1
  • Epidermal Growth Factor
  • ErbB Receptors