Anti-inflammatory and anti-invasive effects of alpha-melanocyte-stimulating hormone in human melanoma cells

α-Melanocyte stimulating hormone (α-MSH) is known to have pleiotrophic functions including pigmentary, anti-inflammatory, antipyretic and immunoregulatory roles in the mammalian body. It is also reported to influence melanoma invasion with levels of α-, β- and γ-MSH correlated clinically with malignant melanoma development, but other studies suggest α-MSH acts to retard invasion. In the present study, we investigated the action of α-MSH on three human melanoma cell lines (HBL, A375-SM and C8161) differing in metastatic potential. α-melanocyte-simulating hormone reduced invasion through fibronectin and also through a human reconstructed skin composite model for the HBL line, and inhibited proinflammatory cytokine-stimulated activation of the NF-κB transcription factor. However, A375-SM and C8161 cells did not respond to α-MSH. Immunofluorescent microscopy and Western blotting identified melanocortin-1 receptor (MC-1R) expression for all three lines and MC-2R on HBL and A375-SM lines. Receptor binding identified a similar affinity for α-MSH for all three lines with the highest number of binding sites on HBL cells. Only the HBL melanoma line demonstrated a detectable cyclic adenosine monophosphate (cAMP) response to α-MSH, although all three lines responded to acute α-MSH addition (+(−)-N6-(2-phenylisopropyl)-adenosine (PIA)) with an elevation in intracellular calcium. The nonresponsive lines displayed MC-1R polymorphisms (C8161, Arg (wt) 151/Cys 151; A375-SM, homozygous Cys 151), whereas the HBL line was wild type. Stable transfection of the C8161 line with wild-type MC-1R produced cells whose invasion was significantly inhibited by α-MSH. From this data, we conclude that α-MSH can reduce melanoma cell invasion and protect cells against proinflammatory cytokine attack in cells with the wild-type receptor (HBL).