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Proteasome inhibitors sensitize colon carcinoma cells to TRAIL-induced apoptosis via enhanced release of smac/DIABLO from the mitochondria

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Pathology & Oncology Research

Abstract

The synergistic interaction between proteasome inhibitors and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a promising approach to induce cell death in tumor cells. However, the molecular and biochemical mechanisms of this synergism have been proven to be cell type specific. We therefore focused our investigation on TRAIL-resistant colon carcinoma cells in this study. DNA fragmentation, mitochondrial membrane depolarization and increased caspase-3-like enzyme activity was exclusively induced only by combined treatment with proteasome inhibitors (epoxomicin, MG132, bortezomib/PS-341) and TRAIL. The expression level of anti-apoptotic proteins (XIAP, survivin, Bcl-2, Bcl-Xl), regulated by NF-κB transcription factor, was not effected by any of these treatments. TRAIL alone induced only partial activation of caspase-3 (p20), while the combination of TRAIL and proteasome inhibition led to the full proteolytic activation of caspase-3 (p17). Only the combination treatment induced marked membrane depolarization and the release of cytochrome c, HtrA2/Omi and Smac/DIABLO. Apoptosis-inducing factor (AIF) was not released in any of these conditions. These results are consistent with a model where the full activation of caspase-3 by caspase-8 is dependent on the release of Smac/DIABLO in response to the combined treatment. This molecular mechanism, independent of the inhibition NF-kB activity, may provide rationale for the combination treatment of colon carcinomas with proteasome inhibitors and recombinant TRAIL or agonistic antibody of TRAIL receptors. (Pathology Oncology Research Vol 12, No 3, 133–142)

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Abbreviations

Ac-DEVD-AMC:

Ac-Asp-Glu-Val-Asp-7-amino-4-methyl-coumarin

z-VAD-fmk:

benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethylketone

z-IETD-fmk:

benzyloxycarbonyl-Ile-Glu(OMe)-Thr-Asp (OMe)-fluoromethylketone

MG132:

ben-zyloxycarbonyl-Leu-Leu-Leu-aldehyde

AIF:

apoptosis-inducing factor

IAP:

inhibitor of apoptosis protein

XIAP:

X-linked IAP

Epo:

epoxomicin

References

  1. Chaudan D, Hideshima T, Mitsiades C, et al: Proteasome inhibitor therapy in multiple myeloma. Mol Cancer Ther 4:686–692, 2005

    Article  Google Scholar 

  2. Kloetzel PM: The proteasome and MHC class I antigen processing. Biochim Biophys Acta 1695:225–233, 2004

    Article  PubMed  CAS  Google Scholar 

  3. Roos-Mattjus P, Sistonen L: The ubiquitin-proteasome pathway. Ann Med 36:285–295, 2004

    Article  PubMed  CAS  Google Scholar 

  4. Sun L, Chen ZJ: The novel functions of ubiquitination in signaling. Curr Opin Cell Biol 16:119–126, 2004

    Article  PubMed  CAS  Google Scholar 

  5. Mani A, Gelmann EP: The ubiquitin-proteasome pathway and its role in cancer. J Clin Oncol 23:4776–4789, 2005

    Article  PubMed  CAS  Google Scholar 

  6. Petak I, Vernes R, Szucs KS, et al: A caspase-8-independent component in TRAIL/Apo-2L-induced cell death in human rhabdomyosarcoma cells. Cell Death Differ 10:729–739, 2003

    Article  PubMed  CAS  Google Scholar 

  7. Petak I, Houghton JA: Shared pathways: death receptors and cytotoxic drugs in cancer therapy. Pathol Oncol Res 7:95–106, 2001

    PubMed  CAS  Google Scholar 

  8. Petak I, Douglas L, Tillman DM, et al: Pediatric rhabdomyo-sarcoma cell lines are resistant to Fas-induced apoptosis and highly sensitive to TRAIL-induced apoptosis. Clin Cancer Res 6:4119–4127, 2000

    PubMed  CAS  Google Scholar 

  9. Zhang L, Fang B: Mechanisms of resistance to TRAIL-induced apoptosis in cancer. Cancer Gene Ther 12:228–237, 2005

    Article  PubMed  CAS  Google Scholar 

  10. Guseva NV, Taghiyev AF, Sturm MT, et al: Tumor necrosis factor-related apoptosis-inducing ligand-mediated activation of mitochondria-associated nuclear factor-kappaB in prostatic carcinoma cell lines. Mol Cancer Res 2:574–584, 2004

    PubMed  CAS  Google Scholar 

  11. Aggarwal BB, Bhardwaj U, Takada Y: Regulation of TRAIL-induced apoptosis by ectopic expression of antiapoptotic factors. Vitam Horm 67:453–483, 2004

    Article  PubMed  CAS  Google Scholar 

  12. Yang LQ, Fang DC, Wang RQ, Yang SM: Effect of NF-kappaB, survivin, Bcl-2 and caspase3 on apoptosis of gastric cancer cells induced by tumor necrosis factor related apoptosis inducing ligand. World J Gastroenterol 10:22–25, 2004

    PubMed  CAS  Google Scholar 

  13. Bortul R, Tazzari PL, Cappellini A, et al: Constitutively active Akt1 protects HL60 leukemia cells from TRAIL-induced apoptosis through a mechanism involving NF-kappaB activation and cFLIP(L) up-regulation. Leukemia 17:379–89, 2003

    Article  PubMed  CAS  Google Scholar 

  14. Ehrhardt H, Fulda S, Schmid I, et al: TRAIL induced survival and proliferation in cancer cells resistant towards TRAIL-induced apoptosis mediated by NF-kappaB. Oncogene 22:3842–3852, 2003

    Article  PubMed  CAS  Google Scholar 

  15. Franco AV, Zhang XD, Van Berkel E, et al: The role of NF-kappa B in TNF-related apoptosis-inducing ligand (TRAIL)-induced apoptosis of melanoma cells. J Immunol 166:5337–5345, 2001

    PubMed  CAS  Google Scholar 

  16. Jeremias I, Kupatt C, Baumann B, et al: Inhibition of nuclear factor kappaB activation attenuates apoptosis resistance in lymphoid cells. Blood 91:4624–4631, 1998

    PubMed  CAS  Google Scholar 

  17. Johnson TR, Stone K, Nikrad M, et al: The proteasome inhibitor PS-341 overcomes TRAIL resistance in Bax and cas-pase 9-negative or Bcl-xL overexpressing cells. Oncogene 22:4953–4963, 2003

    Article  PubMed  CAS  Google Scholar 

  18. Zhu H, Guo W, Zhang L, et al: Proteasome inhibitors-mediated TRAIL resensitization and Bik accumulation. Cancer Biol Ther 4:781–786, 2005

    Article  PubMed  CAS  Google Scholar 

  19. Zhu H, Zhang L, Dong F, et al: Bik/NBK accumulation correlates with apoptosis-induction by bortezomib (PS-341, Velcade) and other proteasome inhibitors. Oncogene 24:4993–4999, 2005

    Article  PubMed  CAS  Google Scholar 

  20. Nikrad M, Johnson T, Puthalalath H, et al: The proteasome inhibitor bortezomib sensitizes cells to killing by death receptor ligand TRAIL via BH3-only proteins Bik and Bim. Mol Cancer Ther 4:443–449, 2005

    PubMed  CAS  Google Scholar 

  21. Schneider P: Production of recombinant TRAIL and TRAIL receptor: Fc chimeric proteins. Methods Enzymol 322:325–3245, 2000

    Article  PubMed  CAS  Google Scholar 

  22. Leverkus M, Sprick MR, Wachter T, et al: Proteasome inhibition results in TRAIL sensitization of primary keratinocytes by removing the resistance-mediating block of effector caspase maturation. Mol Cell Biol 23:777–790, 2003

    Article  PubMed  CAS  Google Scholar 

  23. Chauhan D, Li G, Podar K, et al: Targeting mitochondria to overcome conventional and bortezomib/proteasome inhibitor PS-341 resistance in multiple myeloma (MM) cells. Blood 104:2458–2466, 2004

    Article  PubMed  CAS  Google Scholar 

  24. La Vallee TM, Zhan XH, Johnson MS, et al: 2-methoxyestradiol up-regulates death receptor 5 and induces apoptosis through activation of the extrinsic pathway. Cancer Res 63:468–475, 2003

    Google Scholar 

  25. Creagh EM, Murphy BM, Duriez PJ, et al: Smac/Diablo antagonizes ubiquitin ligase activity of inhibitor of apoptosis proteins. J Biol Chem 279:26906–26914, 2004

    Article  PubMed  CAS  Google Scholar 

  26. Du C, Fang M, Li Y, et al: Smac, a mitochondrial protein that promotes cytochrome c-dependent caspase activation by eliminating IAP inhibition. Cell 102:33–42, 2000

    Article  PubMed  CAS  Google Scholar 

  27. Ni H, Ergin M, Huang Q, et al: Analysis of expression of nuclear factor kappa B (NF-kappa B) in multiple myeloma: downregulation of NF-kappa B induces apoptosis. Br J Haematol 115:279–286, 2001

    Article  PubMed  CAS  Google Scholar 

  28. Berenson JR, Ma HM, Vescio R: The role of nuclear factor-kappaB in the biology and treatment of multiple myeloma. Semin Oncol 28:626–633, 2001

    Article  PubMed  CAS  Google Scholar 

  29. Tillman DM, Izeradjene K, Szucs KS, et al: Rottlerin sensitizes colon carcinoma cells to tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis via uncoupling of the mitochondria independent of protein kinase C. Cancer Res 63:5118–5125, 2003

    PubMed  CAS  Google Scholar 

  30. Ganten TM, Koschny R, Haas TL, et al: Proteasome inhibition sensitizes hepatocellular carcinoma cells, but not human hepatocytes, to TRAIL. Hepatology 42:588–597, 2005

    Article  PubMed  CAS  Google Scholar 

  31. Lashinger LM, Zhu K, Williams SA, et al: Bortezomib abolishes tumor necrosis factor-related apoptosis-inducing ligand resistance via a p21-dependent mechanism in human bladder and prostate cancer cells. Cancer Res 65:4902–4908, 2005

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Üllői út 26., H-1085, Budapest, Hungary

    Katalin Nagy, Helga Barti-Juhász, László Kopper & István Peták MD, PhD

  2. Molecular Pathology Research Group, Joint Research Organization of the Hungarian Academy of Science and Semmelweis University, Budapest

    Rudolf Mihalik & László Kopper

  3. Szentágothai János Knowledge Center, Budapest, Hungary

    Katalin Nagy, László Kopper & István Peták MD, PhD

  4. Department of Oncology and Hematology, University of Modena and Reggio Emilia, Modena, Italy

    Massimo Dominici, Carlotta Spano, Gian Luca Cervo & Pierfranco Conte

  5. Division of Molecular Therapeutics, Department of Hematology-Oncology, St. Jude Children’s Research Hospital, Memphis, TN, USA

    Kinga Székely-Szüts, Kamel Izeradjene, Leslie Douglas, Mike Tillman & Janet A Houghton

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  1. Katalin Nagy
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  2. Kinga Székely-Szüts
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  3. Kamel Izeradjene
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  4. Leslie Douglas
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  5. Mike Tillman
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  6. Helga Barti-Juhász
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  7. Massimo Dominici
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  8. Carlotta Spano
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  9. Gian Luca Cervo
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  10. Pierfranco Conte
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  11. Janet A Houghton
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  12. Rudolf Mihalik
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  13. László Kopper
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  14. István Peták MD, PhD
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Correspondence to István Peták MD, PhD.

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Nagy, K., Székely-Szüts, K., Izeradjene, K. et al. Proteasome inhibitors sensitize colon carcinoma cells to TRAIL-induced apoptosis via enhanced release of smac/DIABLO from the mitochondria. Pathol. Oncol. Res. 12, 133–142 (2006). https://doi.org/10.1007/BF02893359

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  • DOI: https://doi.org/10.1007/BF02893359

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