BH3 mimetic ABT-737 and a proteasome inhibitor synergistically kill melanomas through Noxa-dependent apoptosis

J Invest Dermatol. 2009 Apr;129(4):964-71. doi: 10.1038/jid.2008.327. Epub 2008 Nov 6.

Abstract

The Bcl-2 family is important in modulating sensitivity to anticancer drugs in many cancers, including melanomas. The BH3 mimetic ABT-737 is a potent small molecule inhibitor of the anti-apoptotic proteins Bcl-2/Bcl-X(L)/Bcl-w. In this report, we examined whether ABT-737 is effective in killing melanoma cells in combination with the proteasome inhibitor MG-132, and further evaluated the mechanisms of action. Viability, morphological, and Annexin V apoptosis assays showed that ABT-737 alone exhibited little cytotoxicity, yet it displayed strong synergistic lethality when combined with MG-132. In addition, the detection of Bax/Bak activation indicated that the combination treatment synergistically induced mitochondria-mediated apoptosis. Furthermore, mechanistic analysis revealed that this combination treatment induced expression of the pro-apoptotic protein Noxa- and caspase-dependent degradation of the anti-apoptotic protein, Mcl-1. Finally, siRNA-mediated inhibition of Mcl-1 expression significantly increased sensitivity to ABT-737 in these cells, and knocking down Noxa expression protected the cells from cytotoxicity induced by the combination treatment. These findings demonstrate that ABT-737 combined with MG-132 synergistically induced Noxa-dependent mitochondrial-mediated apoptosis. In summary, this study indicates promising therapeutic potential of targeting anti-apoptotic Bcl-2 family members in treating melanoma, and it validates rational molecular approaches that target anti-apoptotic defenses when developing cancer treatments.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects*
  • Biphenyl Compounds / pharmacology*
  • Caspases / physiology
  • Cell Line, Tumor
  • Cysteine Proteinase Inhibitors / pharmacology*
  • Drug Synergism
  • Humans
  • Leupeptins / pharmacology*
  • Melanoma / drug therapy*
  • Melanoma / pathology
  • Mitochondria / physiology
  • Myeloid Cell Leukemia Sequence 1 Protein
  • Nitrophenols / pharmacology*
  • Piperazines / pharmacology
  • Proteasome Inhibitors*
  • Proto-Oncogene Proteins c-bcl-2 / antagonists & inhibitors*
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / physiology
  • Sulfonamides / pharmacology*

Substances

  • ABT-737
  • Antineoplastic Agents
  • Biphenyl Compounds
  • Cysteine Proteinase Inhibitors
  • Leupeptins
  • Myeloid Cell Leukemia Sequence 1 Protein
  • Nitrophenols
  • PMAIP1 protein, human
  • Piperazines
  • Proteasome Inhibitors
  • Proto-Oncogene Proteins c-bcl-2
  • Sulfonamides
  • Caspases
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde