Suppressing Effect of Na+/Ca2+ Exchanger (NCX) Inhibitors on the Growth of Melanoma Cells

The role of calcium ion (CA²⁺) signaling in tumorigenicity has received increasing attention in melanoma research. Previous CA²⁺ signaling studies focused on CA²⁺ entry routes, but rarely explored the role of CA²⁺ extrusion. Functioning of the Na⁺/CA²⁺ exchanger (NCX) on the plasma membrane is the major way of CA²⁺ extrusion, but very few associations between NCX and melanoma have been reported. Here, we explored whether pharmacological modulation of the NCX could suppress melanoma and promise new therapeutic strategies. Methods included cell viability assay, CA²⁺ imaging, immunoblotting, and cell death analysis. The NCX inhibitors SN-6 and YM-244769 were used to selectively block reverse operation of the NCX. Bepridil, KB-R7943, and CB-DMB blocked either reverse or forward NCX operation. We found that blocking the reverse NCX with SN-6 or YM-244769 (5-100 μM) did not affect melanoma cells or increase cytosolic CA²⁺. Bepridil, KB-R7943, and CB-DMB all significantly suppressed melanoma cells with IC₅₀ values of 3-20 μM. Bepridil and KB-R7943 elevated intracellular CA²⁺ level of melanoma. Bepridil-induced melanoma cell death came from cell cycle arrest and enhanced Apoptosis, which were all attenuated by the CA²⁺ chelator BAPTA-AM. As compared with melanoma, normal melanocytes had lower NCX1 expression and were less sensitive to the cytotoxicity of bepridil. In conclusion, blockade of the forward but not the reverse NCX leads to CA²⁺-related cell death in melanoma and the NCX is a potential drug target for Cancer therapy.