Effects of Protein Kinase Inhibitors on Melanin Production in B16 Melanoma Cells Stimulated via Cyclic AMP-dependent Pathway

B16 Melanoma 세포에서 Protein Kinase 억제제들이 Cyclic AMP 경로를 통한 멜라닌 생성에 미치는 영향

  • 차상복 (중앙대학교 의약식품대학원) ;
  • 조남영 (중앙대학교 의약식품대학원) ;
  • 윤미연 (중앙대학교 의약식품대학원) ;
  • 임혜원 (중앙대학교 의약식품대학원) ;
  • 김경원 (중앙대학교 의약식품대학원) ;
  • 박영미 (중앙대학교 의약식품대학원) ;
  • 이지윤 (중앙대학교 약학대학) ;
  • 이진희 (중앙대학교 약학대학) ;
  • 김창종 (중앙대학교 약학대학)
  • Published : 2003.02.01

Abstract

To investigate the effect of protein kinase on melanin production via cAMP-dependent pathway, we measured the melanin amount and tyrosinase activity in B16 melanoma cells stimulated by alpha-melanocyte stimulating hormone (MSH), forskolin and 8-Br-cAMP. MSH, forskolin and 8-Br-cAMP significantly increased both melanin production and tyrosinase activity in B16 cells. Melanin production and tyrosinase activity by MSH are significantly inhibited by cyclic AMP-dependent protein kinase inhibitor (KT5720) and protein kinase C down-regulation treated with PMA. Bisindolmaleimide (1$\mu$M), protein kinase C inhibitor, significantly inhibited melanin production and tyrosinase activity stimulated by MSH, forskolin and 8-Br-cAMP with the following order of potency: MSH>forskolin>8-Br-cAMP. Tyrosine kinase inhibitor, genistein and DHC, significantly inhibited both, but the inhibitory effect was more potent in 8-Br-cAMP-stimulated B16 cells than MSH-stimulated cells. NFkB inhibitor (parthenolide) significantly inhibited melanin production and tyrosinase activity. Neither melanin production nor tyrosinase activity induced by MSH, forskolin and 8-Br-cAMP were affected by KN-62 (calmodulin-dependent protein kinase II inhibitor), PD098059 (mitogen-activated protein kinase inhibitor, MAPKK) and worthmannin (phosphatidylinositol 3-kinase inhibitor). These results suggest that both protein kinase C and tyrosine kinase are involved in melanin production by cyclic AMP-dependent pathway and NFkB pathway may play an important role in cyclic AMP-dependent melanin production in B16 melanoma cells.

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