Involvement of calpain in melanogenesis of mouse B16 melanoma cells.
In the current study, the involvement of calpain, a cysteine proteinase in the regulation of melanogenesis was examined using mouse B16 melanoma cells. In response to alpha-melanocyte-stimulating hormone (a-MSH), B16 melanoma cells underwent differentiation characterized by increased melanin biosynthesis. The total calapain activity was decreased within 2 h following alpha-MSH-treatment, and restored to the initial level in 6-12 h. To further investigate the involvement of calpain in the regulation of melanogenesis, the effect of calpain inhibitors on alpha-MSH-induced melanogenesis was examined. Inhibition of calpain by either N-acetyl-Leu-Leu-norleucinal (ALLN) or calpastatin (CS) peptide blocked alpha-MSH-induced melanogenesis. The magnitude of inhibition of melanin biosynthesis was well correlated with a decrease in the activity of tyrosinase, a key regulatory enzyme in melanogenesis. Treatment of B16 cells with ALLN caused marked decrease in both tyrosinase protein and mRNA levels. These results indicate that calpain would be involved in the melanogenic signaling by modulating the expression of tyrosinase in mouse B16 melanoma cells.
Ohguchi
Kenji
K
Gifu International Institute of Biotechnology, 1-1 Naka-Fudogaoka, Kakamigahara, Gifu, Japan. kohguchi@giib.or.jp
Akao
Yukihiro
Y
Nozawa
Yoshinori
Y
D000818:Animals; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000494:pharmacology; D002154:Calpain / Q000032:analysis / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006728:Hormones / Q000494:pharmacology; D045563:Indolequinones / Q000032:analysis; D007976:Leupeptins / Q000494:pharmacology; D008543:Melanins / Q000096:biosynthesis; D008544:Melanocytes / Q000378:metabolism*; D008546:Melanoma, Experimental; D051379:Mice; D014442:Monophenol Monooxygenase / Q000032:analysis / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D014407:Tumor Cells, Cultured; D000521:alpha-MSH / Q000494:pharmacology
D002135:Calcium-Binding Proteins; D015853:Cysteine Proteinase Inhibitors; D006728:Hormones; D045563:Indolequinones; D007976:Leupeptins; D008543:Melanins; D012333:RNA, Messenger; C063261:acetylleucyl-leucyl-norleucinal; C001123:dopachrome; D000521:alpha-MSH; C033668:calpastatin; D014442:Monophenol Monooxygenase; D002154:Calpain