Breast Cancer Research and Treatment

, Volume 112, Issue 1, pp 53–62 | Cite as

Inactivation of NF-κB by proteasome inhibition contributes to increased apoptosis induced by histone deacetylase inhibitors in human breast cancer cells

  • Josep Domingo-Domènech
  • Raffaella Pippa
  • Marian Tápia
  • Pere Gascón
  • Oriol Bachs
  • Marta Bosch
Preclinical Study


Histone deacetylase inhibitors (HDACi) are a new class of anticancer agents that cause growth arrest, differentiation and/or apoptosis in many tumor cells. As acetylation regulates the activity of the anti-apoptotic transcription factor NF-κB, we investigated whether the proteasome inhibitor MG-132 would inhibit NF-κB activation and as a consequence potentiate HDACi-dependent apoptosis in breast cancer cells. We observed that the HDACi suberoylanilide hydroxamic acid (SAHA) or trichostatin A (TSA) induced cell death but also enhanced NF-κB-activity. This increase of NF-κB activity was strongly reduced by the addition of MG-132. Moreover, MG-132 potentiates the HDACi-induced cell death that was associated with caspase-3 activation, and PARP cleavage. Induction of the stress related kinases JNK and p38 and the up-regulation of p21 and p27 were also observed after co-treatment of cells with HDACi and MG-132. Disruption of the NF-κB pathway by BAY 11-7085 or IκB-SR mimicked the action of MG-132 in promoting HDACi-induced cell death. Thus, the combined treatment with HDACi and proteasome inhibitors potentiates apoptosis in breast cancer cells representing a novel strategy for breast cancer therapy.


HDAC inhibitor SAHA Proteasome inhibition NF-κB Breast cancer Apoptosis 



This work was supported by Grant SAF 2003–08329 from the Ministerio de Educación y Ciencia of Spain.


  1. 1.
    Kuo MH et al (1998) Roles of histone acetyltransferases and deacetylases in gene regulation. Bioessays 20(8):615–626PubMedCrossRefGoogle Scholar
  2. 2.
    Glozak MA et al (2005) Acetylation and deacetylation of non-histone proteins. Gene 363:15–23Google Scholar
  3. 3.
    Fraga MF et al (2005) Loss of acetylation at Lys16 and trimethylation at Lys20 of histone H4 is a common hallmark of human cancer. Nat Genet 37(4):391–400PubMedCrossRefGoogle Scholar
  4. 4.
    Johnstone RW (2002) Histone-deacetylase inhibitors: novel drugs for the treatment of cancer. Nat Rev Drug Discov 1(4):287–299PubMedCrossRefGoogle Scholar
  5. 5.
    Karin M et al (2004) The IKK NF-kappa B system: a treasure trove for drug development. Nat Rev Drug Discov 3(1):17–26PubMedCrossRefGoogle Scholar
  6. 6.
    Aggarwal BB (2004) Nuclear factor-kappaB: the enemy within. Cancer Cell 6(3):203–208PubMedCrossRefGoogle Scholar
  7. 7.
    Domingo-Domenech J et al (2006) Interleukin 6, a nuclear factor-kappaB target, predicts resistance to docetaxel in hormone-independent prostate cancer and nuclear factor-kappaB inhibition by PS-1145 enhances docetaxel antitumor activity. Clin Cancer Res 12(18):5578–5586PubMedCrossRefGoogle Scholar
  8. 8.
    Chen LF et al (2004) Shaping the nuclear action of NF-kappaB. Nat Rev Mol Cell Biol 5(5):392–401PubMedCrossRefGoogle Scholar
  9. 9.
    Chen LF et al (2002) Acetylation of RelA at discrete sites regulates distinct nuclear functions of NF-kappaB. EMBO J 21(23):6539–6548PubMedCrossRefGoogle Scholar
  10. 10.
    Kiernan R et al (2003) Post-activation turn-off of NF-kappa B-dependent transcription is regulated by acetylation of p65. J Biol Chem 278(4):2758–2766PubMedCrossRefGoogle Scholar
  11. 11.
    Chen L et al (2001) Duration of nuclear NF-kappaB action regulated by reversible acetylation. Science 293(5535):1653–1657CrossRefGoogle Scholar
  12. 12.
    Mayo MW et al (2003) Ineffectiveness of histone deacetylase inhibitors to induce apoptosis involves the transcriptional activation of NF-kappa B through the Akt pathway. J Biol Chem 278(21):18980–18989PubMedCrossRefGoogle Scholar
  13. 13.
    Voorhees PM et al (2003) The proteasome as a target for cancer therapy. Clin Cancer Res 9(17):6316–6325PubMedGoogle Scholar
  14. 14.
    Adams J (2003) The proteasome: structure, function, and role in the cell. Cancer Treat Rev 29(Suppl):13–9Google Scholar
  15. 15.
    Traenckner EB et al (1994) A proteasome inhibitor prevents activation of NF-kappa B and stabilizes a newly phosphorylated form of I kappa B-alpha that is still bound to NF-kappa B. EMBO J 13(22):5433–5441PubMedGoogle Scholar
  16. 16.
    Richardson PG et al (2006) Bortezomib: proteasome inhibition as an effective anticancer therapy. Annu Rev Med 5733–5747Google Scholar
  17. 17.
    Codony-Servat J et al (2006) Differential cellular and molecular effects of bortezomib, a proteasome inhibitor, in human breast cancer cells. Mol Cancer Ther 5(3):665–675PubMedCrossRefGoogle Scholar
  18. 18.
    Yang CH et al (2006) Bortezomib (VELCADE(R)) in metastatic breast cancer: pharmacodynamics, biological effects, and prediction of clinical benefits. Ann Oncol 17(5):813–817PubMedCrossRefGoogle Scholar
  19. 19.
    Venkatraman M et al (2005) Biological and chemical inhibitors of NF-kappaB sensitize SiHa cells to cisplatin-induced apoptosis. Mol Carcinog 44(1):51–59PubMedCrossRefGoogle Scholar
  20. 20.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259):680–685PubMedCrossRefGoogle Scholar
  21. 21.
    Adam E et al (2003) Potentiation of tumor necrosis factor-induced NF-kappa B activation by deacetylase inhibitors is associated with a delayed cytoplasmic reappearance of I kappa B alpha. Mol Cell Biol 23(17):6200–6209PubMedCrossRefGoogle Scholar
  22. 22.
    Adams J (2002) Development of the proteasome inhibitor PS-341. Oncologist 7(1):9–16PubMedCrossRefGoogle Scholar
  23. 23.
    Brown K et al (1995) Control of I kappa B-alpha proteolysis by site-specific, signal-induced phosphorylation. Science 267(5203):1485–1488PubMedCrossRefGoogle Scholar
  24. 24.
    Huang L et al (2000) Suberoylanilide hydroxamic acid as a potential therapeutic agent for human breast cancer treatment. Mol Med 6(10):849–866PubMedGoogle Scholar
  25. 25.
    Cogswell PC et al (2000) Selective activation of NF-kappa B subunits in human breast cancer: potential roles for NF-kappa B2/p52 and for Bcl-3. Oncogene 19(9):1123–1131PubMedCrossRefGoogle Scholar
  26. 26.
    Ghosh S et al (2002) Missing pieces in the NF-kappaB puzzle. Cell 109(Suppl):S81–S96PubMedCrossRefGoogle Scholar
  27. 27.
    Adachi M et al (2004) Synergistic effect of histone deacetylase inhibitors FK228 and m-carboxycinnamic acid bis-hydroxamide with proteasome inhibitors PSI and PS-341 against gastrointestinal adenocarcinoma cells. Clin Cancer Res 10(11):3853–3862PubMedCrossRefGoogle Scholar
  28. 28.
    Giuliano M et al (1999) The apoptotic effects and synergistic interaction of sodium butyrate and MG132 in human retinoblastoma Y79 cells. Cancer Res 59(21):5586–5595PubMedGoogle Scholar
  29. 29.
    Pei XY et al (2004) Synergistic induction of oxidative injury and apoptosis in human multiple myeloma cells by the proteasome inhibitor bortezomib and histone deacetylase inhibitors. Clin Cancer Res 10(11):3839–3852PubMedCrossRefGoogle Scholar
  30. 30.
    Dai Y et al (2005) Blockade of histone deacetylase inhibitor-induced RelA/p65 acetylation and NF-kappaB activation potentiates apoptosis in leukemia cells through a process mediated by oxidative damage, XIAP downregulation, and c-Jun N-terminal kinase 1 activation. Mol Cell Biol 25(13):5429–5444PubMedCrossRefGoogle Scholar
  31. 31.
    Xia Z et al (1995) Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 270(5240):1326–1331PubMedCrossRefGoogle Scholar
  32. 32.
    Kelly WK et al (2005) Drug insight: histone deacetylase inhibitors–development of the new targeted anticancer agent suberoylanilide hydroxamic acid. Nat Clin Pract Oncol 2(3):150–157PubMedCrossRefGoogle Scholar
  33. 33.
    Richon VM et al (2000) Histone deacetylase inhibitor selectively induces p21WAF1 expression and gene-associated histone acetylation. Proc Natl Acad Sci USA 97(18):10014–10019PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2007

Authors and Affiliations

  • Josep Domingo-Domènech
    • 1
  • Raffaella Pippa
    • 2
  • Marian Tápia
    • 1
  • Pere Gascón
    • 1
  • Oriol Bachs
    • 2
  • Marta Bosch
    • 2
  1. 1.Laboratory of Experimental Oncology, Department of Medical OncologyHospital Clínic, Institut d’Investigacions Biomédiques August Pi I Sunyer (IDIBAPS)BarcelonaSpain
  2. 2.Department of Cell Biology and Pathology, Faculty of MedicineUniversity of BarcelonaBarcelonaSpain

Personalised recommendations