Requirement for Phosphatidylinositol 3-Kinase Activity in Epidermal Growth Factor-Stimulated Matrix Metalloproteinase (MMP)-9 Production and Cell Surface Association.

Activation of the epidermal growth factor (EGF) receptor regulates many processes associated with metastasis,including modulation of cell:cell and cell:substrate interactions,production of matrix-degrading proteinases,and cellular migration. We have demonstrated previously that EGF stimulates migration and matrix metalloproteinase (MMP)-9-dependent invasion of ovarian cancer cells. In this study,we compare the roles of EGF-induced phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) activities in regulation of cellular responses associated with ovarian tumor cell metastasis. Inhibition of PI3K and MAPK activity impairs EGF-stimulated cell migration,in vitro invasion,and MMP-9 production. PI3K activity is not required for growth factor disruption of cell:cell junctions,whereas inhibitors of extracellular signal-regulated kinase (ERK)1/ERK2 activation and p38 MAPK activity block EGF-dependent junction dissolution. EGF promotes pro-MMP-9 binding to the cell surface through a mechanism that is independent of extracellular enzyme concentration. Interestingly,inhibition of PI3K activity abolishes EGF-induced cell surface association of pro-MMP-9,whereas inhibitors of MAPKs only partially block the response. These data suggest that EGF receptor activation promotes a PI3K-dependent induction of a cell surface pro-MMP-9 binding component that may facilitate gelatinase-mediated cellular invasion and supports an expanded role for elevated PI3K activity in cellular responses associated with ovarian tumor metastasis. In addition,our findings support the hypothesis that divergent kinase activities regulate distinct cellular events associated with growth factor-induced invasion of ovarian cancer cells.