Abstract
Auranofin (AF) is an organogold FDA-approved drug for treating rheumatism and has been repurposed for several pharmacological applications based on its anti-bacterial, anti-fungal and anti-inflammatory activities. To the best of our knowledge, there has been no study on effects of AF on melanogenesis yet. Hence, in this work, we studied the effect of AF on melanogenesis using B16F10 mouse melanoma cells and validated results in MNT-1 human melanoma cells. Melanogenesis assay was conducted with concentrations of AF determined to be nontoxic in B16F10 cells as well as HaCaT human epidermal cell line for a duration of 48 h, followed by various assays to delineate mechanisms of melanogenesis inhibition. Ultrastructural analysis was conducted to study further if AF affected melanosome maturation and protein levels of a key melanogenic protein, tyrosinase, and the maturation signaling molecule, cyclic adenosine monophosphate (cAMP), was estimated. Our results demonstrate that AF at nontoxic concentrations of 0.25–1 µM significantly inhibited melanin synthesis in a dose-dependent manner with significant inhibition of 32.85% at 1 µM. The study of mechanisms of melanogenesis inhibition revealed that AF inhibited tyrosinase activity in lysates of B16F10 cells but did not show a direct effect on purified mushroom tyrosinase activity or on copper chelation in a cell-free system, nor did it affect levels of B16F10 tyrosinase protein levels. However, AF significantly down-regulated cAMP levels, inhibited cellular ROS and increased number of melanosomes in immature stages, and also exhibited anti-melanogenic activity in B16F10–HaCaT cocultures. Furthermore, AF showed anti-melanogenic efficacy in MNT-1 cell monocultures and cocultures with an inhibition of intracellular tyrosinase activity. In summary, our results demonstrate a proof-of-principle for AF as a depigmenting agent for hyperpigmentation disorders and adjuvant for melanoma therapeutics.
Abbreviations
- AF:
-
Auranofin
- KA:
-
Kojic acid
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- HI-FBS:
-
Heat-inactivated fetal bovine serum
- ROS:
-
Reactive oxygen species
- LDH:
-
Lactate dehydrogenase
- MEM:
-
Minimum essential medium
- ELISA:
-
Enzyme-linked immunosorbent assay
- cAMP:
-
Cyclic adenosine monophosphate
- L-DOPA:
-
3,4-Dihydroxy-l-phenylalanine
- PV:
-
Pyrocatechol violet
References
Alshangiti A, Togher K, Hegarty SV, Sullivan AM, O'Keeffe GW (2019) The dietary flavonoid isoliquiritigenin is a potent cytotoxin for human neuroblastoma cells. Neuro Signal NS20180201
Ando H, Niki Y, Ito M, Akiyama K, Matsui MS, Yarosh DB, Ichihashi M (2012) Melanosomes are transferred from melanocytes to keratinocytes through the processes of packaging, release, uptake, and dispersion. J Invest Dermatol 132:1222–1229. https://doi.org/10.1038/jid.2011.413
Baek SH, Nam IJ, Kwak HS, Kim KC, Lee SH (2015) Cellular anti-melanogenic effects of a euryale ferox seed extract ethyl acetate fraction via the lysosomal degradation machinery. Int J Mol Sci 16:9217–9235. https://doi.org/10.3390/ijms16059217
Berson JF, Harper DC, Tenza D, Raposo G, Marks MS (2001) Pmel17 initiates premelanosome morphogenesis within multivesicular bodies. Mol Biol Cell 12:3451–3464. https://doi.org/10.1091/mbc.12.11.3451
Breathnach AS (1996) Melanin hyperpigmentation of skin: melasma, topical treatment with azelaic acid, and other therapies. Cutis 57:36–45
Chang TS (2009) An updated review of tyrosinase inhibitors. Int J Mol Sci 10:2440–2475. https://doi.org/10.3390/ijms10062440
Chen KG, Leapman RD, Zhang G, Lai B, Valencia JC, Cardarelli CO, Vieira WD, Hearing VJ, Gottesman MM (2009) Influence of melanosome dynamics on melanoma drug sensitivity. JNCI 101:1259–1271
Cheng SL, Liu RH, Sheu JN, Chen ST, Sinchaikul S, Tsay GJ (2007) Toxicogenomics of A375 human malignant melanoma cells treated with arbutin. J Biomed Sci 14:87–105. https://doi.org/10.1007/s11373-006-9130-6
Choi YJ, Rho HS, Baek HS, Hong YD, Joo YH, Shin SS, Kim JM (2014) Synthesis and biological evaluation of kojic acid derivatives as tyrosinase inhibitors. Bull Korean Chem Soc 35:3647–3650
Duval C, Regnier M, Schmidt R (2001) Distinct melanogenic response of human melanocytes in mono-culture, in co-culture with keratinocytes and in reconstructed epidermis, to UV exposure. Pigment Cell Res 14:348–355
Duval C, Smit NP, Kolb AM, Regnier M, Pavel S, Schmidt R (2002) Keratinocytes control the pheo/eumelanin ratio in cultured normal human melanocytes. Pigment Cell Res 15:440–446
Fang D, Dockery P, Weatherhead B (1998) Stereological studies of the effects of alpha-MSH and cAMP on melanosomes in melanoma cells. Pigment Cell Res 11:337–344
Froscio M, Murray AW, Hurst NP (1989) Inhibition of protein kinase C activity by the antirheumatic drug auranofin. Biochem Pharmacol 38:2087–2089
García-Gavín J, González-Vilas D, Fernández-Redondo V, Toribio J (2010) Pigmented contact dermatitis due to kojic acid. A paradoxical side effect of a skin lightener. Contact Dermatitis 62:63–64
Gordon PR, Mansur CP, Gilchrest BA (1989) Regulation of human melanocyte growth, dendricity, and melanization by keratinocyte derived factors. J Invest Dermatol 92:565–572
Griswold DE, Chabot-Fletcher M, Webb EF, Martin L, Hillegass L (1995) Antiinflammatory activity of topical auranofin in arachidonic acid-and phorbol ester-induced inflammation in mice. Drug Dev Res 34:369–375
Hoek K, Rimm DL, Williams KR, Zhao H, Ariyan S, Lin A, Kluger HM, Berger AJ, Cheng E, Trombetta ES, Wu T, Niinobe M, Yoshikawa K, Hannigan GE, Halaban R (2004) Expression profiling reveals novel pathways in the transformation of melanocytes to melanomas. Cancer Res 64:5270–5282. https://doi.org/10.1158/0008-5472.CAN-04-0731
Ismail N, Ismail M, Mazlan M, Latiff LA, Imam MU, Iqbal S, Azmi NH, Ghafar SA, Chan KW (2013) Thymoquinone prevents beta-amyloid neurotoxicity in primary cultured cerebellar granule neurons. Cell Mol Neurobiol 33:1159–1169. https://doi.org/10.1007/s10571-013-9982-z
Joshi PG, Nair N, Begum G, Joshi NB, Sinkar VP, Vora S (2007) Melanocyte-keratinocyte interaction induces calcium signalling and melanin transfer to keratinocytes. Pigment Cell Res 20:380–384. https://doi.org/10.1111/j.1600-0749.2007.00397.x
Jung H, Chung H, Chang SE, Kang DH, Oh ES (2016) FK506 regulates pigmentation by maturing the melanosome and facilitating their transfer to keratinocytes. Pigment Cell Melanoma Res 29:199–209. https://doi.org/10.1111/pcmr.12443
Kooyers T, Westerhof W (2006) Toxicology and health risks of hydroquinone in skin lightening formulations. J Eur Acad Dermatol Venereol 20:777–780
LeBel CP, Ischiropoulos H, Bondy SC (1992) Evaluation of the probe 2', 7'-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem Res Toxicol 5:227–231
Lee C-S, Nam G, Bae I-H, Park J (2019) Whitening efficacy of ginsenoside F1 through inhibition of melanin transfer in cocultured human melanocytes–keratinocytes and three-dimensional human skin equivalent. J Ginseng Res 43:300
Lee WJ, Rhee DY, Bang SH, Kim SY, Won CH, Lee MW, Choi JH, Chang SE (2015) The natural yeast extract isolated by ethanol precipitation inhibits melanin synthesis by modulating tyrosinase activity and downregulating melanosome transfer. Biosci Biotechnol Biochem 79:1504–1511. https://doi.org/10.1080/09168451.2015.1032880
Liang Y-R, Kang S, Deng L, Xiang L-P, Zheng X-Q (2014) Inhibitory effects of (-)-epigallocatechin-3-gallate on melanogenesis in ultraviolet A-induced B16 murine melanoma cell. Trop J Pharm Res 13:1825–1831
Liu JJ, Liu Q, Wei HL, Yi J, Zhao HS, Gao LP (2011) Inhibition of thioredoxin reductase by auranofin induces apoptosis in adriamycin-resistant human K562 chronic myeloid leukemia cells. Pharmazie 66:440–444
Madeira JM, Bajwa E, Stuart MJ, Hashioka S, Klegeris A (2014) Gold drug auranofin could reduce neuroinflammation by inhibiting microglia cytotoxic secretions and primed respiratory burst. J Neuroimmunol 276:71–79. https://doi.org/10.1016/j.jneuroim.2014.08.615
Martinez-Esparza M, Ferrer C, Castells MT, Garcia-Borron JC, Zuasti A (2001) Transforming growth factor beta1 mediates hypopigmentation of B16 mouse melanoma cells by inhibition of melanin formation and melanosome maturation. Int J Biochem Cell Biol 33:971–983
Park HY, Lee J, Gonzalez S, Middelkamp-Hup MA, Kapasi S, Peterson S, Gilchrest BA (2004) Topical application of a protein kinase C inhibitor reduces skin and hair pigmentation. J Invest Dermatol 122:159–166. https://doi.org/10.1046/j.0022-202X.2003.22134.x
Raposo G, Marks MS (2007) Melanosomes—dark organelles enlighten endosomal membrane transport. Nat Rev Mol Cell Biol 8:786
Schallreuter KU, Lemke KR, Hill HZ, Wood JM (1994) Thioredoxin reductase induction coincides with melanin biosynthesis in brown and black guinea pigs and in murine melanoma cells. J Invest Dermatol 103:820–824
Schallreuter KU, Wood J (1987) Azelaic acid as a competitive inhibitor of thioredoxin reductase in human melanoma cells. Cancer Lett 36:297–305
Thangamani S, Maland M, Mohammad H, Pascuzzi PE, Avramova L, Koehler CM, Hazbun TR, Seleem MN (2017) Repurposing approach identifies auranofin with broad spectrum antifungal activity that targets Mia40-Erv1 pathway. Front Cell Infect Microbiol 7:4. https://doi.org/10.3389/fcimb.2017.00004
Thangamani S, Mohammad H, Abushahba MF, Sobreira TJ, Seleem MN (2016) Repurposing auranofin for the treatment of cutaneous staphylococcal infections. Int J Antimicrob Agents 47:195–201. https://doi.org/10.1016/j.ijantimicag.2015.12.016
Wiederhold NP, Patterson TF, Srinivasan A, Chaturvedi AK, Fothergill AW, Wormley FL, Ramasubramanian AK, Lopez-Ribot JL (2017) Repurposing auranofin as an antifungal: In vitro activity against a variety of medically important fungi. Virulence 8:138–142. https://doi.org/10.1080/21505594.2016.1196301
Won YK, Lin CB, Seiberg M, Chen N, Hu Y, Rossetti D, Saliou C, Loy CJ (2014) Galvanic zinc-copper microparticles inhibit melanogenesis via multiple pigmentary pathways. Arch Dermatol Res 306:27–35. https://doi.org/10.1007/s00403-013-1369-y
Wu SY, Wang HM, Wen YS, Liu W, Li PH, Chiu CC, Chen PC, Huang CY, Sheu JH, Wen ZH (2015) 4-(Phenylsulfanyl)butan-2-one suppresses melanin synthesis and melanosome maturation in vitro and in vivo. Int J Mol Sci 16:20240–20257. https://doi.org/10.3390/ijms160920240
Yoon HS, Lee NH, Hyun CG, Shin DB (2015) Differential effects of methoxylated p-coumaric acids on melanoma in B16/F10 cells. Prev Nutr Food Sci 20:73–77. https://doi.org/10.3746/pnf.2015.20.1.73
Acknowledgements
We would like to thank Susan Van Horn for help with TEM sample preparation and imaging and Dr. Michael Marks, University of Pennsylvania for gracious gift of MNT-1 human melanoma cells. This study did not receive any funding in the public, commercial, or not-for-profit sectors.
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SG conceptualized, designed and performed the experiments, analyzed the data, wrote the manuscript and critically revised the manuscript. SRS provided funding for supplies and reviewed the manuscript.
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Goenka, S., Simon, S.R. Organogold drug Auranofin exhibits anti-melanogenic activity in B16F10 and MNT-1 melanoma cells. Arch Dermatol Res 312, 213–221 (2020). https://doi.org/10.1007/s00403-019-01974-1
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DOI: https://doi.org/10.1007/s00403-019-01974-1