BMS-345541 Targets Inhibitor of κB Kinase and Induces Apoptosis in Melanoma: Involvement of Nuclear Factor κB and Mitochondria Pathways

Purpose: Constitutive activation of inhibitor of κB kinase (IKK) confers melanoma resistance to apoptosis and chemotherapy. Whether IKK is able to serve as a therapeutic target in melanoma is unknown. We explored the possibility of exploiting IKK as a therapeutic target in melanoma by using BMS-345541, a novel compound with a highly selective IKKβ inhibitory activity, to trigger melanoma cell apoptosis. Experimental Design: Three human melanoma cell lines (SK-MEL-5, Hs 294T, and A375), all of which have high constitutive IKK activities, served as in vitro and in vivo melanoma models for treatment with BMS-345541. Two known antitumor drugs (temozolomide and bortezomib) were used as parallel controls for evaluation of the therapeutic efficiency and toxicity of BMS-345541. The effects of BMS-345541 on nuclear factor κB (NF-κB) signaling and on the apoptosis machinery were investigated. Results: Inhibition of constitutive IKK activity by BMS-345541 resulted in the reduction of NF-κB activity, CXCL1 chemokine secretion by cultured melanoma cells and melanoma cell survival in vitro and in vivo. The effect of BMS-345541 on tumor cell growth was through mitochondria-mediated apoptosis, based on the release of apoptosis-inducing factor, dissipation of mitochondrial membrane potential, and reduced ratio of B cell lymphoma gene-2 (Bcl-2)/Bcl-associated X protein (Bax) in mitochondria. The BMS-345541 execution of apoptosis was apoptosis-inducing factor–dependent, but largely caspase-independent. Conclusion: BMS-345541 down-regulation of IKK activity results in mitochondria-mediated apoptosis of tumor cells because the programmed cell death machinery in melanoma cells is highly regulated by NF-κB signaling. Therefore, IKK may serve as a potential target for melanoma therapy.

[1]  J. d'Alayer,et al.  Cysteine protease inhibition prevents mitochondrial apoptosis-inducing factor (AIF) release , 2005, Cell Death and Differentiation.

[2]  Francesca Zazzeroni,et al.  Linking JNK signaling to NF-κB: a key to survival , 2004, Journal of Cell Science.

[3]  J. Martinou,et al.  Role of mitochondrial membrane permeabilization in apoptosis and cancer , 2004, Oncogene.

[4]  P. Hersey,et al.  Staurosporine induces apoptosis of melanoma by both caspase-dependent and -independent apoptotic pathways. , 2004, Molecular cancer therapeutics.

[5]  K. Gelmon,et al.  Pharmacodynamic Behavior of Liposomal Antisense Oligonucleotides Targeting Her-2/neu and Vascular Endothelial Growth Factor in an Ascitic MDA435/LCC6 Human Breast Cancer Model , 2004, Cancer biology & therapy.

[6]  G. Kroemer,et al.  The chemopreventive agent N-(4-hydroxyphenyl)retinamide induces apoptosis through a mitochondrial pathway regulated by proteins from the Bcl-2 family , 2003, Oncogene.

[7]  R. Schmid,et al.  Mitogenic and antiapoptotic role of constitutive NF‐κB/Rel activity in pancreatic cancer , 2003, International journal of cancer.

[8]  S. Lowe,et al.  Apoptosis and melanoma chemoresistance , 2003, Oncogene.

[9]  Douglas B. Evans,et al.  Inhibition of constitutive NF-κB activity by IκBαM suppresses tumorigenesis , 2003, Oncogene.

[10]  C. Distelhorst,et al.  Bcl-2 on the Endoplasmic Reticulum Regulates Bax Activity by Binding to BH3-only Proteins* , 2003, The Journal of Biological Chemistry.

[11]  James R. Burke,et al.  BMS-345541 Is a Highly Selective Inhibitor of IκB Kinase That Binds at an Allosteric Site of the Enzyme and Blocks NF-κB-dependent Transcription in Mice* , 2003, The Journal of Biological Chemistry.

[12]  Jean-Claude Martinou,et al.  Mitochondrial release of apoptosis-inducing factor occurs downstream of cytochrome c release in response to several proapoptotic stimuli , 2002, The Journal of cell biology.

[13]  J. Fernandez-Checa,et al.  Ganglioside GD3 Sensitizes Human Hepatoma Cells to Cancer Therapy* , 2002, The Journal of Biological Chemistry.

[14]  A. Hauschild,et al.  Temozolomid als Therapieoption bei Patienten mit fernmetastasiertem Melanom und ungünstiger Prognose , 2002, Der Hautarzt.

[15]  Wayne N. Frankel,et al.  The harlequin mouse mutation downregulates apoptosis-inducing factor , 2002, Nature.

[16]  G. Kroemer,et al.  DNA binding is required for the apoptogenic action of apoptosis inducing factor , 2002, Nature Structural Biology.

[17]  Ann Richmond,et al.  NF-κB, chemokine gene transcription and tumour growth , 2002, Nature Reviews Immunology.

[18]  R. Orlowski,et al.  NF-?B as a therapeutic target in cancer , 2002 .

[19]  Scott W. Lowe,et al.  Apoptosis A Link between Cancer Genetics and Chemotherapy , 2002, Cell.

[20]  B. Wadzinski,et al.  Protein Phosphatase 2A Interacts with and Directly Dephosphorylates RelA* , 2001, The Journal of Biological Chemistry.

[21]  L. Chin,et al.  Induction of melanoma in murine macrophage inflammatory protein 2 transgenic mice heterozygous for inhibitor of kinase/alternate reading frame. , 2001, Cancer research.

[22]  J. Bae,et al.  Underphosphorylated BAD interacts with diverse antiapoptotic Bcl-2 family proteins to regulate apoptosis , 2001, Apoptosis.

[23]  H. Nakayama,et al.  A serine protease, HtrA2, is released from the mitochondria and interacts with XIAP, inducing cell death. , 2001, Molecular cell.

[24]  J. Eberle,et al.  The Bax/Bcl-2 ratio determines the susceptibility of human melanoma cells to CD95/Fas-mediated apoptosis. , 2001, The Journal of investigative dermatology.

[25]  B. Saile.  The bcl, NFκB and p53/p21WAF1 systems are involved in spontaneous apoptosis and in the anti-apoptotic effect of TGF-β or TNF-α on activated hepatic stellate cells , 2001 .

[26]  Xu Luo,et al.  Endonuclease G is an apoptotic DNase when released from mitochondria , 2001, Nature.

[27]  Jingqing Yang,et al.  Constitutive IκB Kinase Activity Correlates with Nuclear Factor-κB Activation in Human Melanoma Cells , 2001 .

[28]  J. Wolchok,et al.  Vaccines for melanoma: translating basic immunology into new therapies. , 2001, The Lancet. Oncology.

[29]  E. Cesarman,et al.  Inhibition of NF-kappaB induces apoptosis of KSHV-infected primary effusion lymphoma cells. , 2000, Blood.

[30]  Bernd R. Binder,et al.  Dynamics of NF κB and IκBα Studied with Green Fluorescent Protein (GFP) Fusion Proteins , 2000, The Journal of Biological Chemistry.

[31]  K. Vousden,et al.  Role of NF-κB in p53-mediated programmed cell death , 2000, Nature.

[32]  A. Ricca,et al.  bcl‐2 over‐expression enhances NF‐κB activity and induces mmp‐9 transcription in human MCF7ADR breast‐cancer cells , 2000, International journal of cancer.

[33]  M. Burdick,et al.  The tumorigenic and angiogenic effects of MGSA/GRO proteins in melanoma , 2000, Journal of leukocyte biology.

[34]  Marty W. Mayo,et al.  NF-κB Induces Expression of the Bcl-2 Homologue A1/Bfl-1 To Preferentially Suppress Chemotherapy-Induced Apoptosis , 1999, Molecular and Cellular Biology.

[35]  E. Androphy,et al.  Human Papillomavirus Type 16 E6-enhanced Susceptibility of L929 Cells to Tumor Necrosis Factor α Correlates with Increased Accumulation of Reactive Oxygen Species* , 1999, The Journal of Biological Chemistry.

[36]  G. Ghosh,et al.  IKAPPABALPHA/NF-KAPPAB COMPLEX , 1999 .

[37]  Ruedi Aebersold,et al.  Molecular characterization of mitochondrial apoptosis-inducing factor , 1999, Nature.

[38]  G. Ghosh,et al.  The Crystal Structure of the IκBα/NF-κB Complex Reveals Mechanisms of NF-κB Inactivation , 1998, Cell.

[39]  Mike Rothe,et al.  IκB Kinase-β: NF-κB Activation and Complex Formation with IκB Kinase-α and NIK , 1997 .

[40]  Matthias Mann,et al.  IKK-1 and IKK-2: Cytokine-Activated IκB Kinases Essential for NF-κB Activation , 1997 .

[41]  Seamus J. Martin,et al.  Suppression of TNF-α-Induced Apoptosis by NF-κB , 1996, Science.

[42]  G. Kroemer,et al.  Inhibitors of permeability transition interfere with the disruption of the mitochondrial transmembrane potential during apoptosis , 1996, FEBS letters.

[43]  D. White,et al.  Interaction of the v-Rel oncoprotein with NF-kappaB and IkappaB proteins: heterodimers of a transformation-defective v-Rel mutant and NF-2 are functional in vitro and in vivo , 1996, Molecular and cellular biology.

[44]  W. Telford,et al.  Rapid quantitation of apoptosis in pure and heterogeneous cell populations using flow cytometry. , 1994, Journal of immunological methods.

[45]  E. Seftor,et al.  Characterization of a highly invasive and spontaneously metastatic human malignant melanoma cell line , 1991, International journal of cancer.

[46]  H. Temin,et al.  Activation of oncogenicity of the c-rel proto-oncogene , 1986, Molecular and cellular biology.

[47]  A. Richmond,et al.  Characterization of autostimulatory and transforming growth factors from human melanoma cells. , 1985, Cancer research.

[48]  A. Hauschild,et al.  [Temozolomide as therapeutic option for patients with metastatic melanoma and poor prognosis]. , 2002, Der Hautarzt; Zeitschrift fur Dermatologie, Venerologie, und verwandte Gebiete.

[49]  E. Kieff,et al.  NF-κB Inhibition in EBV-Transformed Lymphoblastoid Cell Lines , 2002 .

[50]  M J May,et al.  NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses. , 1998, Annual review of immunology.