Synthetic alpha-melanocyte-stimulating hormone (alpha-MSH) was found to bind to the plasma membrane of the HM6A human melanoma cell line, using an immunocytochemical method. When treated with 10(-7) to 10(-9) M alpha-MSH, melanoma cells exhibited an increase of intracellular cyclic adenosine 3':5'-monophosphate, followed by stimulation of tyrosinase activity. Significant inhibition of DNA synthesis measured by [3H]thymidine uptake and inhibition of cell growth was found. A retrovirus expression was detected in the supernatant of HM6A cells as assayed by the KC cell syncytium-forming test. In he presence of 10(-7) M alpha-MSH, the number of syncytium-forming units was increased 15-fold. These results demonstrate that alpha-MSH modulates human melanoma differentiation and virus expression in vitro.