Hirseins inhibit melanogenesis by regulating the gene expressions of Mitf and melanogenesis enzymes

• Published in: Experimental dermatology 2010-05, Vol.19 (5), p.450-457
• Main Authors: Villareal, Myra O., Han, Junkyu, Yamada, Parida, Shigemori, Hideyuki, Isoda, Hiroko
• Format: Article
• Language: English
• Subjects: B16 murine melanoma; Biological and medical sciences; Cell Proliferation - drug effects; Cell Shape - drug effects; Dermatology; Diterpenes - pharmacology; Down-Regulation - drug effects; Down-Regulation - genetics; Enzymes - genetics; Enzymes - metabolism; Gene Expression Regulation - drug effects; Gene Expression Regulation - genetics; Gene Expression Regulation, Enzymologic - drug effects; hirseins; Intramolecular Oxidoreductases - genetics; Medical sciences; Melanins - biosynthesis; melanogenesis; melanogenesis enzymes; Melanoma, Experimental; Microphthalmia-Associated Transcription Factor - genetics; Monophenol Monooxygenase - genetics; Monophenol Monooxygenase - metabolism; Oxidoreductases - genetics; protein kinase C; Protein Kinase C - metabolism; Tumor Cells, Cultured
• ISSN: 0906-6705; 1600-0625
• DOI: 10.1111/j.1600-0625.2009.00964.x

Please cite this paper as: Hirseins inhibit melanogenesis by regulating the gene expressions of Mitf and melanogenesis enzymes. Experimental Dermatology 2009; 19: 450–457. :  Previously, we reported that Thymelaea hirsuta extract has antimelanogenesis effect on B16 murine melanoma cells. The extract was subjected to fractionation, and hirsein A (HA) and hirsein B (HB) were discovered and tested for their ability to regulate melanogenesis in B16 cells. Western blot (WB) analysis was carried out to determine the expression of tyrosinase. Moreover, to elucidate the possible mechanism behind melanogenesis regulation, real‐time PCR using primers for Mitf, Tyr, Trp1 and Dct genes, and protein kinase C (PKC) activity assay were carried out. Results clearly show that 0.1 μm HA and HB significantly reduced the melanin content. This reduction in melanin content was accompanied by reduced tyrosinase expression as detected by WB analysis. There was also a significant decrease in the expression level of Mitf gene in HA‐ and HB‐treated cells. HA down‐regulated the expressions of Tyr, Trp1 and Dct, whereas HB down‐regulated only those of Trp1 and Dct. Interestingly, HB‐treated cells had lower kinase activity than HA‐treated cells indicating a possible difference in the activities of the compounds but with the same mechanism of melanogenesis regulation. We report for the first time that HA and HB can down‐regulate melanogenesis by down‐regulating Mitf gene expression, leading to reduced expressions of Tyr, Trp1 and Dct. The hirseins were also able to reduce the kinase activity, suggesting the possible involvement of PKC in the overall ability of the hirseins to down‐regulate melanogenesis.