Role of p38 MAPK in lupeol-induced B16 2F2 mouse melanoma cell differentiation.

We examined the signaling mechanisms involved in the differentiation-inducing activity of lupeol toward B16 2F2 melanoma cells. alpha-Melanocyte stimulating hormone (alpha-MSH), forskolin and dibutyryl cAMP, which are believed to be cAMP-elevating agents and analogues, enhanced lupeol-induced B16 2F2 cell differentiation. However, H89, an inhibitor of protein kinase A, completely abolished B16-2F2 cell differentiation induced by lupeol. Furthermore, we studied the role of mitogen-activated protein kinases (MAPKs) in lupeol-induced B16 2F2 cell differentiation. U0126, an inhibitor of MAPK kinases, induced B16 2F2 cell differentiation and enhanced the cell differentiation induced by lupeol. However, SB203580, a selective inhibitor of p38 MAPK, completely blocked lupeol-induced B16 2F2 cell differentiation. Western blot analysis revealed that 10 microM lupeol transiently elevated the level of phosphorylation of p38 MAPK. The phosphorylation of p38 MAPK was detected on the addition of 1 microM lupeone, another lupane triterpene, but was not induced by 1 microM lupeol. These results suggested that lupeol induced B16 2F2 cell differentiation through activation of p38 MAPK, and that the structural differences at C-3 of lupane triterpenes played an important role in the activation of p38 MAPK.