Highlights
- •A natural compound harmine decreased melanin synthesis and tyrosinase expression in human MNT-1 melanoma cells and human primary melanocytes.
- •Melanin synthesis was reduced by inhibition of DYRK1A, a harmine target, and enhanced by knockdown of NFATC3, a potential DYRK1A target.
- •Harmine decreases melanin synthesis through regulation of the DYRK1A/NFATC3 pathway.
Abstract
Background
Melanin plays important roles in determining human skin color and protecting human
skin cells against harmful ultraviolet light. However, abnormal hyperpigmentation
in some areas of the skin may become aesthetically unpleasing, resulting in the need
for effective agents or methods to regulate undesirable hyperpigmentation.
Objective
We investigated the effect of harmine, a natural harmala alkaloid belonging to the
beta-carboline family, on melanin synthesis and further explored the signaling pathways
involved in its mechanism of action.
Methods
Human MNT-1 melanoma cells and human primary melanocytes were treated with harmine,
chemical inhibitors, small interfering RNAs, or mammalian expression vectors. Cell
viability, melanin content, and expression of various target molecules were assessed.
Results
Harmine decreased melanin synthesis and tyrosinase expression in human MNT-1 melanoma
cells. Inhibition of DYRK1A, a harmine target, decreased melanin synthesis and tyrosinase
expression. Further studies revealed that nuclear translocation of NFATC3, a potential
DYRK1A substrate, was induced via the harmine/DYRK1A pathway and that NFATC3 knockdown
increased melanin synthesis and tyrosinase expression. Suppression of melanin synthesis
and tyrosinase expression via the harmine/DYRK1A pathway was significantly attenuated
by NFATC3 knockdown. Furthermore, harmine also decreased melanin synthesis and tyrosinase
expression through regulation of NFATC3 in human primary melanocytes.
Conclusion
Our results indicate that harmine decreases melanin synthesis through regulation of
the DYRK1A/NFATC3 pathway and suggest that the DYRK1A/NFATC3 pathway may be a potential
target for the development of depigmenting agents.
Abbreviations:
DYRK1A (dual-specificity tyrosine-phosphorylation-regulated kinase 1A), NFATC (nuclear factor of activated T-cells cytoplasmic), MITF (microphthalmia-associated transcription factor), DOPA (beta-3,4-dihydroxyphenylalanine), TYRP-1 (tyrosinase-related protein-1), TYRP-2 (tyrosinase-related protein-2)Keywords
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Journal of Dermatological ScienceAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Synthesis and physiological implications of melanic pigments.Oncol. Lett. 2019; 17: 4183-4187
- Melanosome transfer to and translocation in the keratinocyte.Exp. Dermatol. 2003; 12: 5-12
- Human skin pigmentation: melanocytes modulate skin color in response to stress.FASEB J. 2007; 21: 976-994
- The protective role of melanin against UV damage in human skin.Photochem. Photobiol. 2008; 84: 539-549
- Mechanisms of ultraviolet light-induced pigmentation.Photochem. Photobiol. 1996; 63: 1-10
- Regulation of human skin pigmentation and responses to ultraviolet radiation.Pigment Cell Res. 2007; 20: 2-13
- The regulation of skin pigmentation.J. Biol. Chem. 2007; 282: 27557-27561
- Physiological factors that regulate skin pigmentation.Biofactors. 2009; 35: 193-199
- Microphthalamia-associated transcription factor: a critical regulator of pigment cell development and survival.Oncogene. 2003; 22: 3035-3041
- Inhibition of the phosphatidylinositol 3-kinase/p70(S6)-kinase pathway induces B16 melanoma cell differentiation.J. Biol. Chem. 1996; 271: 31824-31830
- Microphthalmia associated transcription factor is a target of the phosphatidylinositol-3-kinase pathway.J. Invest. Dermatol. 2003; 121: 831-836
- Delayed ERK activation by ceramide reduces melanin synthesis in human melanocytes.Cell. Signal. 2002; 14: 779-785
- [6]-Shogaol inhibits melanogenesis in B16 mouse melanoma cells through activation of the ERK pathway.Acta Pharmacol. Sin. 2013; 34: 289-294
- Melanocyte-specific expression of the human tyrosinase promoter: activation by the microphthalmia gene product and role of the initiator.Mol. Cell. Biol. 1994; 14: 7996-8006
- Functional analysis of microphthalmia-associated transcription factor in pigment cell-specific transcription of the human tyrosinase family genes.J. Biol. Chem. 1997; 272: 503-509
- Microphthalmia gene product as a signal transducer in cAMP-induced differentiation of melanocytes.J. Cell Biol. 1998; 142: 827-835
- Different cis-acting elements are involved in the regulation of TRP1 and TRP2 promoter activities by cyclic AMP: pivotal role of M boxes (GTCATGTGCT) and of microphthalmia.Mol. Cell. Biol. 1998; 18: 694-702
- Harmine specifically inhibits protein kinase DYRK1A and interferes with neurite formation.FEBS J. 2009; 276: 6324-6337
- Harmine is an ATP-competitive inhibitor for dual-specificity tyrosine phosphorylation-regulated kinase 1A (Dyrk1A).Arch. Biochem. Biophys. 2011; 507: 212-218
- Inhibition of monoamine oxidase A by beta-carboline derivatives.Arch. Biochem. Biophys. 1997; 337: 137-142
- Specific inhibition of cyclin-dependent kinases and cell proliferation by harmine.Biochem. Biophys. Res. Commun. 2004; 317: 128-132
- A high-throughput chemical screen reveals that harmine-mediated inhibition of DYRK1A increases human pancreatic beta cell replication.Nat. Med. 2015; 21: 383-388
- The small molecule harmine is an antidiabetic cell-type-specific regulator of PPARgamma expression.Cell Metab. 2007; 5: 357-370
- The small molecule harmine regulates NFATc1 and Id2 expression in osteoclast progenitor cells.Bone. 2011; 49: 264-274
- Harmine promotes osteoblast differentiation through bone morphogenetic protein signaling.Biochem. Biophys. Res. Commun. 2011; 409: 260-265
- Novel chondrogenic and chondroprotective effects of the natural compound harmine.Biochimie. 2013; 95: 374-381
- IL-4 inhibits the melanogenesis of normal human melanocytes through the JAK2-STAT6 signaling pathway.J. Invest. Dermatol. 2013; 133: 528-536
- Secreted frizzled-related protein 2 (sFRP2) functions as a melanogenic stimulator; the role of sFRP2 in UV-induced hyperpigmentary disorders.J. Invest. Dermatol. 2016; 136: 236-244
- Epidermal growth factor-induced matrix metalloproteinase-1 expression is negatively regulated by p38 MAPK in human skin fibroblasts.J. Dermatol. Sci. 2011; 64: 134-141
- Expression profiling reveals novel pathways in the transformation of melanocytes to melanomas.Cancer Res. 2004; 64: 5270-5282
- A model for melanosome biogenesis based on the purification and analysis of early melanosomes.Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 10698-10703
- Functions of adaptor protein (AP)-3 and AP-1 in tyrosinase sorting from endosomes to melanosomes.Mol. Biol. Cell. 2005; 16: 5356-5372
- BLOC-1 interacts with BLOC-2 and the AP-3 complex to facilitate protein trafficking on endosomes.Mol. Biol. Cell. 2006; 17: 4027-4038
- Genome-wide siRNA-based functional genomics of pigmentation identifies novel genes and pathways that impact melanogenesis in human cells.PLoS Genet. 2008; 4: e1000298
- ULK1 regulates melanin levels in MNT-1 cells independently of mTORC1.PLoS One. 2013; 8: e75313
- Development of a novel selective inhibitor of the Down syndrome-related kinase Dyrk1A.Nat. Commun. 2010; 1: 86
- The specificities of protein kinase inhibitors: an update.Biochem. J. 2003; 371: 199-204
- DYRK family of protein kinases: evolutionary relationships, biochemical properties, and functional roles.FASEB J. 2011; 25: 449-462
- Function and regulation of Dyrk1A: towards understanding Down syndrome.Cell. Mol. Life Sci. 2009; 66: 3235-3240
- Activation, regulation, and inhibition of DYRK1A.FEBS J. 2011; 278: 246-256
- NFAT gene family in inflammation and cancer.Curr. Mol. Med. 2013; 13: 543-554
- Partners in transcription: NFAT and AP-1.Oncogene. 2001; 20: 2476-2489
- Transcription factors of the NFAT family: regulation and function.Annu. Rev. Immunol. 1997; 15: 707-747
- Alternative splicing and expression of human and mouse NFAT genes.Genomics. 2008; 92: 279-291
- A genome-wide Drosophila RNAi screen identifies DYRK-family kinases as regulators of NFAT.Nature. 2006; 441: 646-650
- Downregulation of NFAT2 promotes melanogenesis in B16 melanoma cells.Anat. Cell Biol. 2010; 43: 303-309
Article info
Publication history
Published online: May 15, 2021
Accepted:
May 12,
2021
Received in revised form:
May 11,
2021
Received:
January 8,
2021
Identification
Copyright
© 2021 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.
