ACTIVATION STATUS AND FUNCTION OF THE VLA-4 (ALPHA-4-BETA-1) INTEGRINEXPRESSED ON HUMAN-MELANOMA CELL-LINES

We have examined the functional status of the VLA-4/alpha 4 beta 1 int egrin in a panel of human melanoma cell lines, focusing on the ability of cells expressing alpha 4 beta 1 to mediate adhesion to the alpha 4 -specific ligands CS-1 peptide and VCAM-1. All melanoma cells expressi ng alpha 4 beta 1 (8 of 10 lines examined) were capable of adhering to these specific ligands in adhesion assays, whereas 2 cell lines (HMB2 and VUP) which lacked surface alpha 4 were unable to do so, Adherence of different melanoma cell lines to VCAM-1 was relatively uniform and not susceptible to upregulation with known integrin-activating factor s, such as manganese ions, phorbol ester and activating monoclonal ant ibody (mAb) TS2/16, Cell adhesion to CS-1 peptide, however, varied acc ording to cell surface receptor density and, in some cases, could be u p-regulated by integrin-activating factors, Adhesion of SK23 cells to CS-1 peptide was increased by all 3 activating stimuli, whereas for al l other melanoma cells an increase was obtained only by the use of TS2 /16 mAb. Our data indicate not only an unusually low activation state of alpha 4 beta 1 in SK23 cells but also heterogeneity in the activati ng capacity of the various stimuli. Moreover, a protein kinase C-depen dent role in alpha 4 beta 1 activity was suggested by adhesion assays carried out in the presence of the protein kinase C inhibitor calphost in C, which considerably reduced adhesion to CS-1 peptide. (C) 1997 Wi ley-Liss, Inc.