Investigational New Drugs

, Volume 30, Issue 4, pp 1289–1301

The redox antimalarial dihydroartemisinin targets human metastatic melanoma cells but not primary melanocytes with induction of NOXA-dependent apoptosis

  • Christopher M. Cabello
  • Sarah D. Lamore
  • Warner B. BairIII
  • Shuxi Qiao
  • Sara Azimian
  • Jessica L. Lesson
  • Georg T. Wondrak


Recent research suggests that altered redox control of melanoma cell survival, proliferation, and invasiveness represents a chemical vulnerability that can be targeted by pharmacological modulation of cellular oxidative stress. The endoperoxide artemisinin and semisynthetic artemisinin-derivatives including dihydroartemisinin (DHA) constitute a major class of antimalarials that kill plasmodium parasites through induction of iron-dependent oxidative stress. Here, we demonstrate that DHA may serve as a redox chemotherapeutic that selectively induces melanoma cell apoptosis without compromising viability of primary human melanocytes. Cultured human metastatic melanoma cells (A375, G361, LOX) were sensitive to DHA-induced apoptosis with upregulation of cellular oxidative stress, phosphatidylserine externalization, and activational cleavage of procaspase 3. Expression array analysis revealed DHA-induced upregulation of oxidative and genotoxic stress response genes (GADD45A, GADD153, CDKN1A, PMAIP1, HMOX1, EGR1) in A375 cells. DHA exposure caused early upregulation of the BH3-only protein NOXA, a proapototic member of the Bcl2 family encoded by PMAIP1, and genetic antagonism (siRNA targeting PMAIP1) rescued melanoma cells from apoptosis indicating a causative role of NOXA-upregulation in DHA-induced melanoma cell death. Comet analysis revealed early DHA-induction of genotoxic stress accompanied by p53 activational phosphorylation (Ser 15). In primary human epidermal melanocytes, viability was not compromised by DHA, and oxidative stress, comet tail moment, and PMAIP1 (NOXA) expression remained unaltered. Taken together, these data demonstrate that metastatic melanoma cells display a specific vulnerability to DHA-induced NOXA-dependent apoptosis and suggest feasibility of future anti-melanoma intervention using artemisinin-derived clinical redox antimalarials.


Malignant melanoma Dihydroartemisinin PMAIP1 Reactive oxygen species Oxidative stress Apoptosis 


  1. 1.
    Fruehauf JP, Trapp V (2008) Reactive oxygen species: an Achilles' heel of melanoma? Expert Rev Anticancer Ther 8:1751–1757CrossRefPubMedGoogle Scholar
  2. 2.
    Garbe C, Eigentler TK, Keilholz U, Hauschild A, Kirkwood JM (2011) Systematic review of medical treatment in melanoma: current status and future prospects. Oncologist 16:5–24CrossRefPubMedGoogle Scholar
  3. 3.
    Meyskens FL Jr, Farmer P, Fruehauf JP (2001) Redox regulation in human melanocytes and melanoma. Pigment Cell Res 14:148–154CrossRefPubMedGoogle Scholar
  4. 4.
    Wittgen HG, van Kempen LC (2007) Reactive oxygen species in melanoma and its therapeutic implications. Melanoma Res 17:400–409CrossRefPubMedGoogle Scholar
  5. 5.
    Govindarajan B, Sligh JE, Vincent BJ, Li M, Canter JA, Nickoloff BJ, Rodenburg RJ, Smeitink JA, Oberley L, Zhang Y, Slingerland J, Arnold RS, Lambeth JD, Cohen C, Hilenski L, Griendling K, Martinez-Diez M, Cuezva JM, Arbiser JL (2007) Overexpression of Akt converts radial growth melanoma to vertical growth melanoma. J Clin Invest 117:719–729CrossRefPubMedGoogle Scholar
  6. 6.
    Fried L, Arbiser JL (2008) The reactive oxygen-driven tumor: relevance to melanoma. Pigment Cell Melanoma Res 21:117–122CrossRefPubMedGoogle Scholar
  7. 7.
    Gidanian S, Mentelle M, Meyskens FL Jr, Farmer PJ (2008) Melanosomal damage in normal human melanocytes induced by UVB and metal uptake–a basis for the pro-oxidant state of melanoma. Photochem Photobiol 84:556–564CrossRefPubMedGoogle Scholar
  8. 8.
    Laurent A, Nicco C, Chereau C, Goulvestre C, Alexandre J, Alves A, Levy E, Goldwasser F, Panis Y, Soubrane O, Weill B, Batteux F (2005) Controlling tumor growth by modulating endogenous production of reactive oxygen species. Cancer Res 65:948–956PubMedGoogle Scholar
  9. 9.
    Cabello CM, Bair WB 3rd, Wondrak GT (2007) Experimental therapeutics: targeting the redox Achilles heel of cancer. Curr Opin Investig Drugs 8:1022–1037PubMedGoogle Scholar
  10. 10.
    Trachootham D, Alexandre J, Huang P (2009) Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach? Nat Rev Drug Discov 8:579–591CrossRefPubMedGoogle Scholar
  11. 11.
    Wondrak GT (2009) Redox-directed cancer therapeutics: molecular mechanisms and opportunities. Antioxid Redox Signal 11:3013–3069CrossRefPubMedGoogle Scholar
  12. 12.
    Efferth T (2007) Willmar Schwabe Award 2006: antiplasmodial and antitumor activity of artemisinin–from bench to bedside. Planta Med 73:299–309CrossRefPubMedGoogle Scholar
  13. 13.
    Berger TG, Dieckmann D, Efferth T, Schultz ES, Funk JO, Baur A, Schuler G (2005) Artesunate in the treatment of metastatic uveal melanoma–first experiences. Oncol Rep 14:1599–1603PubMedGoogle Scholar
  14. 14.
    Chen H, Sun B, Pan S, Jiang H, Sun X (2009) Dihydroartemisinin inhibits growth of pancreatic cancer cells in vitro and in vivo. Anticancer Drugs 20:131–140CrossRefPubMedGoogle Scholar
  15. 15.
    Mercer AE, Maggs JL, Sun XM, Cohen GM, Chadwick J, O'Neill PM, Park BK (2007) Evidence for the involvement of carbon-centered radicals in the induction of apoptotic cell death by artemisinin compounds. J Biol Chem 282:9372–9382CrossRefPubMedGoogle Scholar
  16. 16.
    Ramacher M, Umansky V, Efferth T (2009) Effect of artesunate on immune cells in ret-transgenic mouse melanoma model. Anticancer Drugs 20:910–917CrossRefPubMedGoogle Scholar
  17. 17.
    Buommino E, Baroni A, Canozo N, Petrazzuolo M, Nicoletti R, Vozza A, Tufano MA (2009) Artemisinin reduces human melanoma cell migration by down-regulating alpha V beta 3 integrin and reducing metalloproteinase 2 production. Invest New Drugs 27:412–418CrossRefPubMedGoogle Scholar
  18. 18.
    Stockwin LH, Han B, Yu SX, Hollingshead MG, ElSohly MA, Gul W, Slade D, Galal AM, Newton DL, Bumke MA (2009) Artemisinin dimer anticancer activity correlates with heme-catalyzed reactive oxygen species generation and endoplasmic reticulum stress induction. Int J Cancer 125:1266–1275CrossRefPubMedGoogle Scholar
  19. 19.
    Wondrak GT (2007) NQO1-activated phenothiazinium redox cyclers for the targeted bioreductive induction of cancer cell apoptosis. Free Radic Biol Med 43:178–190CrossRefPubMedGoogle Scholar
  20. 20.
    Cabello CM, Bair WB 3rd, Bause AS, Wondrak GT (2009) Antimelanoma activity of the redox dye DCPIP (2,6-dichlorophenolindophenol) is antagonized by NQO1. Biochem Pharmacol 78:344–354CrossRefPubMedGoogle Scholar
  21. 21.
    Lamore SD, Cabello CM, Wondrak GT (2010) The topical antimicrobial zinc pyrithione is a heat shock response inducer that causes DNA damage and PARP-dependent energy crisis in human skin cells. Cell Stress Chaperones 15:309–322CrossRefPubMedGoogle Scholar
  22. 22.
    Lamore SD, Qiao S, Horn D, Wondrak GT (2010) Proteomic identification of cathepsin B and nucleophosmin as novel UVA-targets in human skin fibroblasts. Photochem Photobiol 86:1307–1317CrossRefPubMedGoogle Scholar
  23. 23.
    Cabello CM, Lamore SD, Bair WB, Davis AL, Azimian SM, Wondrak GT (2011) DCPIP (2,6-dichlorophenolindophenol) as a genotype-directed redox chemotherapeutic targeting NQO1*2 breast carcinoma. Free Radic Res 45:276–292CrossRefPubMedGoogle Scholar
  24. 24.
    Mills JC, Stone NL, Erhardt J, Pittman RN (1998) Apoptotic membrane blebbing is regulated by myosin light chain phosphorylation. J Cell Biol 140:627–636CrossRefPubMedGoogle Scholar
  25. 25.
    Qin JZ, Ziffra J, Stennett L, Bodner B, Bonish BK, Chaturvedi V, Bennett F, Pollock PM, Trent JM, Hendrix MJ, Rizzo P, Miele L, Nickoloff BJ (2005) Proteasome inhibitors trigger NOXA-mediated apoptosis in melanoma and myeloma cells. Cancer Res 65:6282–6293CrossRefPubMedGoogle Scholar
  26. 26.
    Fernandez Y, Verhaegen M, Miller TP, Rush JL, Steiner P, Opipari AW Jr, Lowe SW, Soengas MS (2005) Differential regulation of noxa in normal melanocytes and melanoma cells by proteasome inhibition: therapeutic implications. Cancer Res 65:6294–6304CrossRefPubMedGoogle Scholar
  27. 27.
    Yu KS, Lee Y, Kim CM, Park EC, Choi J, Lim DS, Chung YH, Koh SS (2010) The protease inhibitor, elafin, induces p53-dependent apoptosis in human melanoma cells. Int J Cancer 127:1308–1320CrossRefPubMedGoogle Scholar
  28. 28.
    Kinner A, Wu W, Staudt C, Iliakis G (2008) {gamma}-H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin. Nucleic Acids ResGoogle Scholar
  29. 29.
    Rong JJ, Hu R, Song XM, Ha J, Lu N, Qi Q, Tao L, You QD, Guo QL (2010) Gambogic acid triggers DNA damage signaling that induces p53/p21(Waf1/CIP1) activation through the ATR-Chk1 pathway. Cancer Lett 296:55–64CrossRefPubMedGoogle Scholar
  30. 30.
    Mercer AE, Copple IM, Maggs JL, O’Neill PM, Park BK (2011) The role of heme and the mitochondrion in the chemical and molecular mechanisms of mammalian cell death induced by the artemisinin antimalarials. J Biol Chem 286:987–996CrossRefPubMedGoogle Scholar
  31. 31.
    Handrick R, Ontikatze T, Bauer KD, Freier F, Rubel A, Durig J, Belka C, Jendrossek V (2010) Dihydroartemisinin induces apoptosis by a Bak-dependent intrinsic pathway. Mol Cancer Ther 9:2497–2510CrossRefPubMedGoogle Scholar
  32. 32.
    Efferth T, Briehl MM, Tome ME (2003) Role of antioxidant genes for the activity of artesunate against tumor cells. Int J Oncol 23:1231–1235PubMedGoogle Scholar
  33. 33.
    Li PC, Lam E, Roos WP, Zdzienicka MZ, Kaina B, Efferth T (2008) Artesunate derived from traditional Chinese medicine induces DNA damage and repair. Cancer Res 68:4347–4351CrossRefPubMedGoogle Scholar
  34. 34.
    Moore JC, Lai H, Li JR, Ren RL, McDougall JA, Singh NP, Chou CK (1995) Oral administration of dihydroartemisinin and ferrous sulfate retarded implanted fibrosarcoma growth in the rat. Cancer Lett 98:83–87PubMedGoogle Scholar
  35. 35.
    Richardson DR, Kalinowski DS, Lau S, Jansson PJ, Lovejoy DB (2009) Cancer cell iron metabolism and the development of potent iron chelators as anti-tumour agents. Biochim Biophys Acta 1790:702–717CrossRefPubMedGoogle Scholar
  36. 36.
    Yang WS, Stockwell BR (2008) Synthetic lethal screening identifies compounds activating iron-dependent, nonapoptotic cell death in oncogenic-RAS-harboring cancer cells. Chem Biol 15:234–245CrossRefPubMedGoogle Scholar
  37. 37.
    Tuma RS (2011) Getting around PLX4032: studies turn up unusual mechanisms of resistance to melanoma drug. J Natl Cancer Inst 103(170–171):177Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Christopher M. Cabello
    • 1
  • Sarah D. Lamore
    • 1
  • Warner B. BairIII
    • 1
  • Shuxi Qiao
    • 1
  • Sara Azimian
    • 1
  • Jessica L. Lesson
    • 1
  • Georg T. Wondrak
    • 1
  1. 1.College of Pharmacy & Arizona Cancer Center, Department of Pharmacology and ToxicologyUniversity of ArizonaTucsonUSA

Personalised recommendations