latrunculin-b and Fibrosarcoma

Integrins are highly regulated receptors that can function in both cell-substrate and cell-cell adhesion. We have found that the activating anti-beta1 mAb, 12G10, can specifically and rapidly induce both cell-substrate and cell-cell adhesion of HT-1080 human fibrosarcoma and other cell types. Binding of mAb 12G10 induced clustering of cell-surface integrins, and the preferential localization of beta1 integrins expressing the 12G10 epitope at cell-cell adhesion sites. Fab fragments of mAb 12G10 induced HT-1080 cell-cell adhesion as effectively as did intact antibodies, suggesting that integrin clustering was not due to direct antibody crosslinking. Latrunculin B, an inhibitor of F-actin polymerization, inhibited cell-cell adhesion but not the clustering of integrins. Results from a novel, two-color cell-cell adhesion assay suggested that nonactivated cells can bind to activated cells and that integrin activation-induced HT-1080 cell-cell adhesion minimally requires the interaction of activated alpha2beta1 with nonactivated alpha3beta1. These findings suggest that HT-1080 cell-cell adhesion induced by integrin activation require a signaling process involving integrin clustering and the subsequent organization of the cytoskeleton. Integrin activation could therefore play a key role in cell-cell adhesion.

Topics: Actins; Antibodies, Monoclonal; Bridged Bicyclo Compounds, Heterocyclic; Cadherins; Cations, Divalent; Cell Adhesion; Collagen; Dose-Response Relationship, Immunologic; Epitopes; Fibronectins; Fibrosarcoma; Humans; Hybridomas; Immunohistochemistry; Integrin beta1; Laminin; Microscopy, Confocal; Models, Biological; Signal Transduction; Thiazoles; Thiazolidines; Tumor Cells, Cultured