Marisa M Fernández1, Fátima Ferragut2, Víctor M Cárdenas Delgado2, Candelaria Bracalente2, Alicia I Bravo3, Alejandro J Cagnoni4, Myriam Nuñez5, Luciano G Morosi6, Héctor R Quinta2, María V Espelt2, María F Troncoso2, Carlota Wolfenstein-Todel2, Karina V Mariño4, Emilio L Malchiodi1, Gabriel A Rabinovich7, María T Elola8. 1. Institute of Studies in Humoral Immunology, University of Buenos Aires (UBA) and National Council Research (CONICET), Microbiology, Immunology and Biotechnology Department, School of Pharmacy and Biochemistry, University of Buenos Aires (UBA), Buenos Aires, Argentina. 2. Institute of Biochemistry and Biophysics (IQUIFIB), UBA-CONICET, Biological Chemistry Department, School of Pharmacy and Biochemistry, UBA, Buenos Aires, Argentina. 3. Molecular Pathology Department, "Eva Perón" HIGA Hospital, Buenos Aires, Argentina. 4. Laboratory of Functional and Molecular Glycomics, Institute of Biology and Experimental Medicine (IBYME), CONICET, Buenos Aires, Argentina. 5. Department of Mathematics and Statistics, School of Pharmacy and Biochemistry, UBA, Buenos Aires, Argentina. 6. Laboratory of Functional and Molecular Glycomics, Institute of Biology and Experimental Medicine (IBYME), CONICET, Buenos Aires, Argentina; Laboratory of Immunopathology, IBYME, CONICET, Buenos Aires, Argentina. 7. Laboratory of Immunopathology, IBYME, CONICET, Buenos Aires, Argentina; Faculty of Exact and Natural Sciences, UBA, Buenos Aires, Argentina. 8. Institute of Biochemistry and Biophysics (IQUIFIB), UBA-CONICET, Biological Chemistry Department, School of Pharmacy and Biochemistry, UBA, Buenos Aires, Argentina. Electronic address: mt_elola@yahoo.com.
Abstract
BACKGROUND: We previously demonstrated that the activated leukocyte cell adhesion molecule (ALCAM/CD166) can interact with galectin-8 (Gal-8) in endothelial cells. ALCAM is a member of the immunoglobulin superfamily that promotes homophilic and heterophilic cell-cell interactions. Gal-8 is a "tandem-repeat"-type galectin, known as a matricellular protein involved in cell adhesion. Here, we analyzed the physical interaction between both molecules in breast cancer cells and the functional relevance of this phenomenon. METHODS: We performed binding assays by surface plasmon resonance to study the interaction between Gal-8 and the recombinant glycosylated ALCAM ectodomain or endogenous ALCAM from MDA-MB-231 breast cancer cells. We also analyzed the binding of ALCAM-silenced or control breast cancer cells to immobilized Gal-8 by SPR. In internalization assays, we evaluated the influence of Gal-8 on ALCAM surface localization. RESULTS: We showed that recombinant glycosylated ALCAM and endogenous ALCAM from breast carcinoma cells physically interacted with Gal-8 in a glycosylation-dependent fashion displaying a differential behavior compared to non-glycosylated ALCAM. Moreover, ALCAM-silenced breast cancer cells exhibited reduced binding to Gal-8 relative to control cells. Importantly, exogenously added Gal-8 provoked ALCAM segregation, probably trapping this adhesion molecule at the surface of breast cancer cells. CONCLUSIONS: Our data indicate that Gal-8 interacts with ALCAM at the surface of breast cancer cells through glycosylation-dependent mechanisms. GENERAL SIGNIFICANCE: A novel heterophilic interaction between ALCAM and Gal-8 is demonstrated here, suggesting its physiologic relevance in the biology of breast cancer cells.
BACKGROUND: We previously demonstrated that the activated leukocyte cell adhesion molecule (ALCAM/CD166) can interact with galectin-8 (Gal-8) in endothelial cells. ALCAM is a member of the immunoglobulin superfamily that promotes homophilic and heterophilic cell-cell interactions. Gal-8 is a "tandem-repeat"-type galectin, known as a matricellular protein involved in cell adhesion. Here, we analyzed the physical interaction between both molecules in breast cancer cells and the functional relevance of this phenomenon. METHODS: We performed binding assays by surface plasmon resonance to study the interaction between Gal-8 and the recombinant glycosylated ALCAM ectodomain or endogenous ALCAM from MDA-MB-231 breast cancer cells. We also analyzed the binding of ALCAM-silenced or control breast cancer cells to immobilized Gal-8 by SPR. In internalization assays, we evaluated the influence of Gal-8 on ALCAM surface localization. RESULTS: We showed that recombinant glycosylated ALCAM and endogenous ALCAM from breast carcinoma cells physically interacted with Gal-8 in a glycosylation-dependent fashion displaying a differential behavior compared to non-glycosylated ALCAM. Moreover, ALCAM-silenced breast cancer cells exhibited reduced binding to Gal-8 relative to control cells. Importantly, exogenously added Gal-8 provoked ALCAM segregation, probably trapping this adhesion molecule at the surface of breast cancer cells. CONCLUSIONS: Our data indicate that Gal-8 interacts with ALCAM at the surface of breast cancer cells through glycosylation-dependent mechanisms. GENERAL SIGNIFICANCE: A novel heterophilic interaction between ALCAM and Gal-8 is demonstrated here, suggesting its physiologic relevance in the biology of breast cancer cells.
Authors: Kamil Wdowiak; Tomasz Francuz; Enrique Gallego-Colon; Natalia Ruiz-Agamez; Marcin Kubeczko; Iga Grochoła; Jerzy Wojnar Journal: Int J Mol Sci Date: 2018-01-10 Impact factor: 5.923
Authors: Henri-François Renard; François Tyckaert; Cristina Lo Giudice; Thibault Hirsch; Cesar Augusto Valades-Cruz; Camille Lemaigre; Massiullah Shafaq-Zadah; Christian Wunder; Ruddy Wattiez; Ludger Johannes; Pierre van der Bruggen; David Alsteens; Pierre Morsomme Journal: Nat Commun Date: 2020-03-19 Impact factor: 14.919