Contribution of sialic acids to integrin α5β1 functioning in melanoma cells.

Abstract

PURPOSE:

To establish the relationship between sialylation of integrin α5β1 and possible alteration in the function of α5β1 receptor in melanoma cells.

MATERIALS AND METHODS:

Integrin α5β1 was isolated from primary WM115 (RGP/VGP-like phenotype) and metastatic WM266-4 (lymph node metastasis) cells via affinity chromatography. Integrin α5β1 sialylation and the shift in relative masses of the enzymatically desialylated subunits were confirmed by confocal microscopy and SDS-PAGE, respectively. The ELISA assay was performed to evaluate sialic acid (SA) influence on integrin α5β1 binding to fibronectin (FN). Cell invasion was investigated by the Transwell invasion assay. The effect of neuraminidases treatment on melanoma cells was assessed by flow cytometry using Maackia amurensis and Sambucus nigra lectins.

RESULTS:

Both subunits of integrin α5β1 were found to be more abundantly sialylated in primary than in metastatic cells. The removal of SA had no effect on the purified integrin α5β1 binding to FN. Although metastatic cells underwent more pronounced desialylation than primary cells, invasion of primary WM115 cells was more dependent on the presence of α2-3 linked SA than it was in the case of metastatic WM266-4 cells. In both melanoma cell lines not only integrin α5β1 was involved in invasion, however simultaneous desialylation and usage of anti-integrin α5β1 antibodies resulted in lower invasion abilities of primary WM115 cells.

CONCLUSIONS:

Our data suggest that in primary melanoma cells integrin α5β1 action is more likely dependent on its glycosylation profile, i.e. the presence of SA residues, which influence (decreased) their invasion properties and may facilitate malignant melanoma progression.

• Authors: Ewa Kolasińska, Marcelina E Janik, Anna Lityńska, Małgorzata Przybyło
• Journal: Advances in medical sciences (Adv Med Sci) Vol. 64 Issue 2 Pg. 267-273 (Sep 2019) ISSN: 1898-4002 [Electronic] Netherlands
• PMID: 30844664 (Publication Type: Journal Article)
• Copyright: Copyright © 2019 Medical University of Bialystok. Published by Elsevier B.V. All rights reserved.

Chemical References

• Integrin alpha5beta1
• Sialic Acids

Topics

• Animals
• Cell Line, Tumor
• Chromatography, Affinity
• Electrophoresis, Polyacrylamide Gel
• Glycosylation
• Humans
• Integrin alpha5beta1 (metabolism)
• Melanoma (metabolism)
• Mice
• Microscopy, Confocal
• Sialic Acids (pharmacology)
• Skin Neoplasms (metabolism)