MSH Receptors and the Response of Human A375 Melanoma Cells to Interleukin-1β

Abstract

α-Melanocyte-stimulating hormone (α-MSH, α-melanotropin) has been shown to be an inhibitory factor in many immunologic and inflammatory processes involving the cytokine interleukin-1 (IL-1). As the mechanism of the interaction between IL-1 and α-MSH at the receptor level is unknown, we have studied the role of MC1 melanocortin receptors in two variants of the human melanoma cell line A375 differing in their sensitivity to the cytostatic effects of IL-1β. Both IL-1 sensitive (A375r-) and resistant cells (A375r+) carry specific high affinity receptors for IL-1, albeit their concentration is 10-fold higher in A375r+ cells. In A375r- cells, MC1 receptors are absent or below the level for reliable detection in the binding assay. Conversion of A375r- to A375r+ cells by prolonged culture in medium not depleted of endotoxin led to the appearance of MC1 receptors (KD 0.4 ± 0.123 nmol/l; 608 ± 134 receptors/cell). Stable transfection of A375r- cells with the human MC1 receptor did not, however, render them resistant to the cytostatic effect of IL-1β on concomitant treatment with α-MSH or result in the production of IL-6 on treatment with IL-1β Therefore, the presence of MC1 receptors on the surface of A375 cells or their binding to α-MSH does not seem to be a factor in cytokine resistance or IL-6 secretion. No interaction between IL-1β and α-MSH could be demonstrated at the cellular level in this melanoma cell line.