Organogold drug Auranofin exhibits anti-melanogenic activity in B16F10 and MNT-1 melanoma cells

  • Shilpi GoenkaEmail author
  • Sanford R. Simon
Concise Communication


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.


Auranofin Drug repurposing Anti-melanogenic Tyrosinase Melanosome maturation 





Kojic acid


Dulbecco’s modified Eagle’s medium


Heat-inactivated fetal bovine serum


Reactive oxygen species


Lactate dehydrogenase


Minimum essential medium


Enzyme-linked immunosorbent assay


Cyclic adenosine monophosphate




Pyrocatechol violet



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.

Author contributions

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.

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.

Human or animal rights statement

This article does not contain any studies involving animals or human participants.

Supplementary material

403_2019_1974_MOESM1_ESM.docx (463 kb)
Supplementary file1 (DOCX 463 kb)


  1. 1.
    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 NS20180201Google Scholar
  2. 2.
    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. CrossRefGoogle Scholar
  3. 3.
    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. CrossRefGoogle Scholar
  4. 4.
    Berson JF, Harper DC, Tenza D, Raposo G, Marks MS (2001) Pmel17 initiates premelanosome morphogenesis within multivesicular bodies. Mol Biol Cell 12:3451–3464. CrossRefGoogle Scholar
  5. 5.
    Breathnach AS (1996) Melanin hyperpigmentation of skin: melasma, topical treatment with azelaic acid, and other therapies. Cutis 57:36–45Google Scholar
  6. 6.
    Chang TS (2009) An updated review of tyrosinase inhibitors. Int J Mol Sci 10:2440–2475. CrossRefGoogle Scholar
  7. 7.
    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 101:1259–1271CrossRefGoogle Scholar
  8. 8.
    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. CrossRefGoogle Scholar
  9. 9.
    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 35:3647–3650CrossRefGoogle Scholar
  10. 10.
    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 14:348–355CrossRefGoogle Scholar
  11. 11.
    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 15:440–446CrossRefGoogle Scholar
  12. 12.
    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 11:337–344CrossRefGoogle Scholar
  13. 13.
    Froscio M, Murray AW, Hurst NP (1989) Inhibition of protein kinase C activity by the antirheumatic drug auranofin. Biochem Pharmacol 38:2087–2089CrossRefGoogle Scholar
  14. 14.
    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 62:63–64CrossRefGoogle Scholar
  15. 15.
    Gordon PR, Mansur CP, Gilchrest BA (1989) Regulation of human melanocyte growth, dendricity, and melanization by keratinocyte derived factors. J Invest Dermatol 92:565–572CrossRefGoogle Scholar
  16. 16.
    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 34:369–375CrossRefGoogle Scholar
  17. 17.
    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. CrossRefGoogle Scholar
  18. 18.
    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. CrossRefGoogle Scholar
  19. 19.
    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. Google Scholar
  20. 20.
    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. CrossRefGoogle Scholar
  21. 21.
    Kooyers T, Westerhof W (2006) Toxicology and health risks of hydroquinone in skin lightening formulations. J Eur Acad Dermatol Venereol 20:777–780Google Scholar
  22. 22.
    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 5:227–231CrossRefGoogle Scholar
  23. 23.
    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 43:300CrossRefGoogle Scholar
  24. 24.
    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. CrossRefGoogle Scholar
  25. 25.
    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 13:1825–1831CrossRefGoogle Scholar
  26. 26.
    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 66:440–444Google Scholar
  27. 27.
    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. CrossRefGoogle Scholar
  28. 28.
    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 33:971–983CrossRefGoogle Scholar
  29. 29.
    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. CrossRefGoogle Scholar
  30. 30.
    Raposo G, Marks MS (2007) Melanosomes—dark organelles enlighten endosomal membrane transport. Nat Rev Mol Cell Biol 8:786CrossRefGoogle Scholar
  31. 31.
    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 103:820–824CrossRefGoogle Scholar
  32. 32.
    Schallreuter KU, Wood J (1987) Azelaic acid as a competitive inhibitor of thioredoxin reductase in human melanoma cells. Cancer Lett 36:297–305CrossRefGoogle Scholar
  33. 33.
    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. Google Scholar
  34. 34.
    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. CrossRefGoogle Scholar
  35. 35.
    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. CrossRefGoogle Scholar
  36. 36.
    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. CrossRefGoogle Scholar
  37. 37.
    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. CrossRefGoogle Scholar
  38. 38.
    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. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biomedical EngineeringStony Brook UniversityStony BrookUSA
  2. 2.Department of PathologyStony Brook UniversityStony BrookUSA
  3. 3.Department of Biochemistry and Cellular BiologyStony Brook UniversityStony BrookUSA

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