Epithelial growth factor (EGF) via its receptor EGFR promotes epithelial ovarian cancer (EOC) metastasis, though the effect of inhibition of this pathway is minimal in clinical trials. We have identified the cell surface glycoprotein CUB domain containing protein 1 (CDCP1) as a key regulator of EGF/EGFR signaling pathways. We show that signaling via EGF/EGFR induces migration of Caov3 and OVCA420 EOC cells with concomitant up-regulation of CDCP1 mRNA and protein. Consistent with a role in cell migration CDCP1 relocates from cell-cell junctions to punctate structures on filopodia following activation of EGFR. Significantly, disruption of CDCP1, either by silencing or the use of a function blocking antibody, efficiently reduces EGF/EGFR-induced migration of these EOC cells. We also show that up-regulation of CDCP1 is inhibited by pharmacological agents blocking ERK signaling indicating that the MAPKinase pathway is required downstream of EGF in the EOC microenvironment to up-regulate CDCP1. The γ2 chain of laminin localises to the ovarian epithelial basal lamina and EOC. Over-expression of intracellular laminin γ2 is seen in the cytoplasm of cells at the invasion front in several carcinomas, and free γ2 promotes cell migration, suggesting its role in tumour progression. We cultured EOC cells on top of Matrigel to mimic the ovarian stromal microenvironment. Laminin γ2 was observed around more 3-dimensional (3D) spheroids formed by the Caov3 and OVCA420 cells treated with the CDCP1 blocking antibody than mouse IgG treated control cells. Additionally, a significant decrease in the number and size of 3D spheroids was observed in treated Caov3 and OVCA420. Our data indicate that CDCP1 inhibition reduced the growth of these cells and may have a role in restoring the basal membrane localisation of laminin. Together, these results suggest that CDCP1 mediates EGF/EGFR induced cell signaling and may be a potential target to limit the EOC metastasis.