Exchange protein directly activated by cyclic AMP increases melanoma cell migration by a Ca2+-dependent mechanism

Melanoma has a poor prognosis due to its strong metastatic ability. Although CA(2+) plays a major role in cell migration, little is known about the role of CA(2+) in melanoma cell migration. We recently found that the exchange protein directly activated by cyclic AMP (Epac) increases melanoma cell migration via a heparan sulfate-related mechanism. In addition to this mechanism, we also found that Epac regulates melanoma cell migration by a CA(2+)-dependent mechanism. An Epac agonist increased CA(2+) in several different melanoma cell lines but not in melanocytes. Ablation of Epac1 with short hairpin RNA inhibited the Epac agonist-induced CA(2+) elevation, suggesting the critical role of Epac1 in CA(2+) homeostasis in melanoma cells. Epac-induced CA(2+) elevation was negated by the inhibition of Phospholipase C (PLC) and inositol triphosphate (IP(3)) receptor. Furthermore, Epac-induced cell migration was reduced by the inhibition of PLC or IP(3) receptor. These data suggest that Epac activates CA(2+) release from the endoplasmic reticulum via the PLC/IP(3) receptor pathway, and this CA(2+) elevation is involved in Epac-induced cell migration. Actin assembly was increased by Epac-induced CA(2+), suggesting the involvement of actin in Epac-induced cell migration. In human melanoma specimens, mRNA expression of Epac1 was higher in metastatic melanoma than in primary melanoma, suggesting a role for Epac1 in melanoma metastasis. In conclusion, our findings reveal that Epac is a potential target for the suppression of melanoma cell migration, and, thus, the development of metastasis.