Endothelin receptor B (ET(B)R) is a
G-protein-coupled receptor overexpressed in
melanoma, blood, and lymphatic endothelial cells. Given that aberrant signal transduction can be mediated through cross talk between receptors, here, we explore the functional relationship between ET(B)R and the
vascular endothelial growth factor receptor (VEGFR)-3 system and how this cross talk might influence the aggressive behavior of
melanoma cells. The expression of
VEGFR-3 and its
ligands,
VEGF-C and
VEGF-D, significantly increased after activating ET(B)R by ET-1 in primary and metastatic
melanoma cell lines. These effects, similarly to those induced by
hypoxia, were mediated by
hypoxia-inducible factor (HIF)-1α and HIF-2α. ET-1 caused the phosphorylation of
VEGFR-3, which was accompanied by the activation of the downstream signaling molecules, such as MAPK and AKT. Inhibition of c-Src activity or silencing of the scaffold
protein β-arrestin-1 reduced ET-1-induced
VEGFR-3 phosphorylation, demonstrating that, upon ET-1 stimulus, β-arrestin-1 is involved with c-Src in the ET(B)R-mediated
VEGFR-3 transactivation. Moreover, ET-1 in combination with
VEGF-C further increased
VEGFR-3, MAPK, and AKT phosphorylation and markedly promoted cell migration and vasculogenic mimicry. Dual inhibition of ET(B)R and
VEGFR-3 was required for the effective inhibition of these effects, as well as for
VEGFR-3 phosphorylation, demonstrating that ET(B)R cross talk with
VEGFR-3 enhances cell plasticity and motility. Finally, in
melanoma xenografts, ET(B)R antagonist inhibited
tumor growth and the activation of the
VEGF-C/VEGFR-3 axis, indicating that targeting ET(B)R may improve
melanoma treatment acting directly or indirectly by impairing ET(B)R cross talk with
VEGFR-3.