Cell migration of ovarian tumoral cells is essential for cell dissemination and for invasion of the submesothelial extracellular matrix (ECM). We have conducted a study of the migratory properties of an ovarian adenocarcinoma cell line (IGROV1) by using 2 distinct methods for the evaluation of cell migration. We found that in a short‐term transfilter migration assay, IGROV1 cells migrated toward vitronectin, fibronectin, type IV collagen and laminin in an integrin‐dependent manner. When migration was evaluated in a wound healing assay, the restitution of the wounded area was stimulated solely by added, exogenous vitronectin and was almost totally dependent on αvβ3 integrin function. Moreover, we demonstrated that αvβ3 was localized in focal contacts restricted to the leading edge of migrating cells, whereas vitronectin notably localized with actin stress fibers and cortical actin. On the other hand, several kinase inhibitors were found to impede migration of IGROV1 induced by vitronectin. It thus appears that αvβ3–vitronectin interactions lead to the activation of multiple signaling pathway including activation of protein kinase C, phosphatidyl‐inositol‐3‐phosphate kinase and protein tyrosine kinase. The “αvβ3–vitronectin system” is therefore essential to the migration of human ovarian carcinoma cells.Int. J. Cancer 80:285–294, 1999. © 1999 Wiley‐Liss, Inc.