Modulation of endothelial cell morphogenesis in vitro by MMP-9 during glial-endothelial cell interactions

The purpose of this study was to investigate the roles of matrix metallopro teinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in t he formation of capillary structures by human brain microvascular endotheli al cells cocultured with SNB19 glioblastoma cells. Unstimulated cocultures did not form capillaries and produce MMP-9 but stimulation with the protein kinase C (PKC) activator 4-phorbol-12-myristate 13-acetate (PMA) produced MMP-9 and capillary networks. Addition of recombinant MMP-9 increased capil lary formation. Anti-MMP-9 antibodies, TIMP-1, the synthetic MMPs inhibitor Batimastat (BB-94), and the PKC inhibitor calphostin-C all reduced MMP-9 a ctivity and capillary network formation in these cocultures. Cytochalasin-D in the presence of PMA suppressed MMP-9 expression and capillary formation , but colchicine-B had no such effect. Finally, PMA-induced MMP-9 expressio n and capillary formation were inhibited by the MEKK-specific inhibitor PD9 8059. These results suggest that MMP-9 is important in endothelial cell mor phogenesis and the formation of capillaries in glial/endothelial cocultures in vitro.