Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

A pivotal role for Mcl-1 in Bortezomib-induced apoptosis

Oncogene volume 27, pages 721–731 (2008)Cite this article

Abstract

Bortezomib is a proteasome inhibitor for the treatment of relapsed/refractory multiple myeloma (MM). Mechanisms of resistance to Bortezomib are undefined. Myeloid cell leukemia-1 (Mcl-1) is an antiapoptotic protein, which protects tumor cells against spontaneous and chemotherapy-induced apoptosis. In MM, specific downregulation of Mcl-1 induces apoptosis. Here, we examined the role of Mcl-1 in Bortezomib- and doxorubicin-induced apoptosis. We demonstrate that Bortezomib, but not doxorubicin, triggers caspase-dependent generation of a 28 kDa Mcl-1-fragment, in several MM cell lines, including MM.1S cells. Conversely, transient transfection of MM.1S cells with a previously reported 28 kDa Mcl-1128–350 fragment, but not with the Mcl-11–127 fragment, induces apoptosis. Therefore, both downregulation of full-length antiapoptotic Mcl-1, as well as Bortezomib-induced generation of Mcl-1128–350 cleaved protein, contribute to MM cell apoptosis. To verify further these findings, we next compared effects triggered by Bortezomib, doxorubicin and melphalan in Mcl-1wt/wt and Mcl-1Δ/null murine embryonic fibroblasts (MEFs). Our results show that Bortezomib, but not doxorubicin or melphalan, triggers Mcl-1 cleavage in Mcl-1wt/wt, but not Mcl-1Δ/null MEFs and induces sub-G1 phase cells; caspase-3 and -9, and PARP cleavage as well as morphological signs of apoptosis. Taken together, these results support an important role of Mcl-1 and a Mcl-1 fragment in Bortezomib-induced cell death in general, and in MM in particular. To prevent relapse of MM in patients treated with Bortezomib, we therefore recommend the combination of Bortezomib with agents that induce MM cell death independent of Mcl-1.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  • Bae J, Leo CP, Hsu SY, Hsueh AJ . (2000). MCL-1S, a splicing variant of the antiapoptotic BCL-2 family member MCL-1, encodes a proapoptotic protein possessing only the BH3 domain. J Biol Chem 275: 25255–25261.

    Article  CAS  PubMed  Google Scholar 

  • Bataille R, Harousseau JL . (1997). Multiple myeloma. N Engl J Med 336: 1657–1664.

    Article  CAS  PubMed  Google Scholar 

  • Chauhan D, Hideshima T, Mitsiades C, Richardson P, Anderson KC . (2005). Proteasome inhibitor therapy in multiple myeloma. Mol Cancer Ther 4: 686–692.

    Article  CAS  PubMed  Google Scholar 

  • Chauhan D, Li G, Podar K, Hideshima T, Mitsiades C, Schlossman R et al. (2004). Targeting mitochondria to overcome conventional and Bortezomib/proteasome inhibitor PS-341 resistance in multiple myeloma (MM) cells. Blood 104: 2458–2466.

    Article  CAS  PubMed  Google Scholar 

  • Chauhan D, Uchiyama H, Akbarali Y, Urashima M, Yamamoto K, Libermann TA et al. (1996). Multiple myeloma cell adhesion-induced interleukin-6 expression in bone marrow stromal cells involves activation of NF-kappa B. Blood 87: 1104–1112.

    CAS  PubMed  Google Scholar 

  • Chen L, Willis SN, Wei A, Smith BJ, Fletcher JI, Hinds MG et al. (2005). Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function. Mol Cell 17: 393–403.

    Article  CAS  PubMed  Google Scholar 

  • Craig RW . (2002). MCL1 provides a window on the role of the BCL2 family in cell proliferation, differentiation and tumorigenesis. Leukemia 16: 444–454.

    Article  CAS  PubMed  Google Scholar 

  • Derenne S, Monia B, Dean NM, Taylor JK, Rapp MJ, Harousseau JL et al. (2002). Antisense strategy shows that Mcl-1 rather than Bcl-2 or Bcl-x(L) is an essential survival protein of human myeloma cells. Blood 100: 194–199.

    Article  CAS  PubMed  Google Scholar 

  • Domina AM, Smith JH, Craig RW . (2000). Myeloid cell leukemia 1 is phosphorylated through two distinct pathways, one associated with extracellular signal-regulated kinase activation and the other with G2/M accumulation or protein phosphatase 1/2A inhibition. J Biol Chem 275: 21688–21694.

    Article  CAS  PubMed  Google Scholar 

  • Domina AM, Vrana JA, Gregory MA, Hann SR, Craig RW . (2004). MCL1 is phosphorylated in the PEST region and stabilized upon ERK activation in viable cells, and at additional sites with cytotoxic okadaic acid or taxol. Oncogene 23: 5301–5315.

    Article  CAS  PubMed  Google Scholar 

  • Gewirtz DA . (1999). A critical evaluation of the mechanisms of action proposed for the antitumor effects of the anthracycline antibiotics adriamycin and daunorubicin. Biochem Pharmacol 57: 727–741.

    Article  CAS  PubMed  Google Scholar 

  • Gomez-Bougie P, Bataille R, Amiot M . (2004). The imbalance between Bim and Mcl-1 expression controls the survival of human myeloma cells. Eur J Immunol 34: 3156–3164.

    Article  CAS  PubMed  Google Scholar 

  • Han J, Goldstein LA, Gastman BR, Froelich CJ, Yin XM, Rabinowich H . (2004). Degradation of Mcl-1 by granzyme B: implications for Bim-mediated mitochondrial apoptotic events. J Biol Chem 279: 22020–22029.

    Article  CAS  PubMed  Google Scholar 

  • Hideshima T, Chauhan D, Hayashi T, Akiyama M, Mitsiades N, Mitsiades C et al. (2003a). Proteasome inhibitor PS-341 abrogates IL-6 triggered signaling cascades via caspase-dependent downregulation of gp130 in multiple myeloma. Oncogene 22: 8386–8393.

    Article  CAS  PubMed  Google Scholar 

  • Hideshima T, Mitsiades C, Akiyama M, Hayashi T, Chauhan D, Richardson P et al. (2003b). Molecular mechanisms mediating antimyeloma activity of proteasome inhibitor PS-341. Blood 101: 1530–1534.

    Article  CAS  PubMed  Google Scholar 

  • Hideshima T, Richardson P, Chauhan D, Palombella VJ, Elliott PJ, Adams J et al. (2001). The proteasome inhibitor PS-341 inhibits growth, induces apoptosis, and overcomes drug resistance in human multiple myeloma cells. Cancer Res 61: 3071–3076.

    CAS  PubMed  Google Scholar 

  • Hurley LH . (2002). DNA and its associated processes as targets for cancer therapy. Nat Rev Cancer 2: 188–200.

    Article  CAS  PubMed  Google Scholar 

  • Inoshita S, Takeda K, Hatai T, Terada Y, Sano M, Hata J et al. (2002). Phosphorylation and inactivation of myeloid cell leukemia 1 by JNK in response to oxidative stress. J Biol Chem 277: 43730–43734.

    Article  CAS  PubMed  Google Scholar 

  • Jang YM, Kendaiah S, Drew B, Phillips T, Selman C, Julian D et al. (2004). Doxorubicin treatment in vivo activates caspase-12 mediated cardiac apoptosis in both male and female rats. FEBS Lett 577: 483–490.

    Article  CAS  PubMed  Google Scholar 

  • Kozopas KM, Yang T, Buchan HL, Zhou P, Craig RW . (1993). MCL1, a gene expressed in programmed myeloid cell differentiation, has sequence similarity to BCL2. Proc Natl Acad Sci USA 90: 3516–3520.

    Article  CAS  PubMed  Google Scholar 

  • LeBlanc R, Catley LP, Hideshima T, Lentzsch S, Mitsiades CS, Mitsiades N et al. (2002). Proteasome inhibitor PS-341 inhibits human myeloma cell growth in vivo and prolongs survival in a murine model. Cancer Res 62: 4996–5000.

    CAS  PubMed  Google Scholar 

  • Le Gouill S, Podar K, Amiot M, Hideshima T, Chauhan D, Ishitsuka K et al. (2004a). VEGF induces Mcl-1 up-regulation and protects multiple myeloma cells against apoptosis. Blood 104: 2886–2892.

    Article  CAS  PubMed  Google Scholar 

  • Le Gouill S, Podar K, Harousseau JL, Anderson KC . (2004b). Mcl-1 regulation and its role in multiple myeloma. Cell Cycle 3: 1259–1262.

    Article  CAS  PubMed  Google Scholar 

  • Leu JI, Dumont P, Hafey M, Murphy ME, George DL . (2004). Mitochondrial p53 activates Bak and causes disruption of a Bak-Mcl1 complex. Nat Cell Biol 6: 443–450.

    Article  CAS  PubMed  Google Scholar 

  • Maurer U, Charvet C, Wagman AS, Dejardin E, Green DR . (2006). Glycogen synthase kinase-3 regulates mitochondrial outer membrane permeabilization and apoptosis by destabilization of MCL-1. Mol Cell 21: 749–760.

    Article  CAS  PubMed  Google Scholar 

  • Michels J, O'Neill JW, Dallman CL, Mouzakiti A, Habens F, Brimmell M et al. (2004). Mcl-1 is required for Akata6 B-lymphoma cell survival and is converted to a cell death molecule by efficient caspase-mediated cleavage. Oncogene 23: 4818–4827.

    Article  CAS  PubMed  Google Scholar 

  • Nijhawan D, Fang M, Traer E, Zhong Q, Gao W, Du F et al. (2003). Elimination of Mcl-1 is required for the initiation of apoptosis following ultraviolet irradiation. Genes Dev 17: 1475–1486.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Opferman JT . (2006). Unraveling MCL-1 degradation. Cell Death Differ 13: 1260–1262.

    Article  CAS  PubMed  Google Scholar 

  • Opferman JT, Iwasaki H, Ong CC, Suh H, Mizuno S, Akashi K et al. (2005). Obligate role of anti-apoptotic MCL-1 in the survival of hematopoietic stem cells. Science 307: 1101–1104.

    Article  CAS  PubMed  Google Scholar 

  • Opferman JT, Letai A, Beard C, Sorcinelli MD, Ong CC, Korsmeyer SJ . (2003). Development and maintenance of B and T lymphocytes requires antiapoptotic MCL-1. Nature 426: 671–676.

    Article  CAS  PubMed  Google Scholar 

  • Orlowski RZ, Voorhees PM, Garcia RA, Hall MD, Kudrik FJ, Allred T et al. (2005). Phase 1 trial of the proteasome inhibitor Bortezomib and pegylated liposomal doxorubicin in patients with advanced hematologic malignancies. Blood 105: 3058–3065.

    Article  CAS  PubMed  Google Scholar 

  • Pei XY, Dai Y, Grant S . (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: 3839–3852.

    Article  CAS  PubMed  Google Scholar 

  • Perez-Galan P, Roue G, Villamor N, Montserrat E, Campo E, Colomer D . (2006). The proteasome inhibitor Bortezomib induces apoptosis in mantle-cell lymphoma through generation of ROS and NOXA activation independent of p53 status. Blood 107: 257–264.

    Article  CAS  PubMed  Google Scholar 

  • Podar K, Catley LP, Tai YT, Shringarpure R, Carvalho P, Hayashi T et al. (2004a). GW654652, the pan-inhibitor of VEGF receptors, blocks the growth and migration of multiple myeloma cells in the bone marrow microenvironment. Blood 103: 3474–3479.

    Article  CAS  PubMed  Google Scholar 

  • Podar K, Shringarpure R, Tai YT, Simoncini M, Sattler M, Ishitsuka K et al. (2004b). Caveolin-1 is required for vascular endothelial growth factor-triggered multiple myeloma cell migration and is targeted by Bortezomib. Cancer Res 64: 7500–7506.

    Article  CAS  PubMed  Google Scholar 

  • Puthier D, Bataille R, Amiot M . (1999). IL-6 up-regulates Mcl-1 in human myeloma cells through JAK/STAT rather than ras/MAP kinase pathway. Eur J Immunol 29: 3945–3950.

    Article  CAS  PubMed  Google Scholar 

  • Qin JZ, Ziffra J, Stennett L, Bodner B, Bonish BK, Chaturvedi V et al. (2005). Proteasome inhibitors trigger NOXA-mediated apoptosis in melanoma and myeloma cells. Cancer Res 65: 6282–6293.

    Article  CAS  PubMed  Google Scholar 

  • Raje N, Kumar S, Hideshima T, Roccaro A, Ishitsuka K, Yasui H et al. (2005). Seliciclib (CYC202 or R-roscovitine), a small-molecule cyclin-dependent kinase inhibitor, mediates activity via down-regulation of Mcl-1 in multiple myeloma. Blood 106: 1042–1047.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Richardson PG, Barlogie B, Berenson J, Singhal S, Jagannath S, Irwin D et al. (2003). A phase 2 study of Bortezomib in relapsed, refractory myeloma. N Engl J Med 348: 2609–2617.

    Article  CAS  PubMed  Google Scholar 

  • Rinkenberger JL, Horning S, Klocke B, Roth K, Korsmeyer SJ . (2000). Mcl-1 deficiency results in peri-implantation embryonic lethality. Genes Dev 14: 23–27.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Roccaro AM, Hideshima T, Raje N, Kumar S, Ishitsuka K, Yasui H et al. (2006). Bortezomib mediates antiangiogenesis in multiple myeloma via direct and indirect effects on endothelial cells. Cancer Res 66: 184–191.

    Article  CAS  PubMed  Google Scholar 

  • Weng C, Li Y, Xu D, Shi Y, Tang H . (2005). Specific cleavage of Mcl-1 by caspase-3 in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in Jurkat leukemia T cells. J Biol Chem 280: 10491–10500.

    Article  CAS  PubMed  Google Scholar 

  • Willis SN, Chen L, Dewson G, Wei A, Naik E, Fletcher JI et al. (2005). Proapoptotic Bak is sequestered by Mcl-1 and Bcl-xL, but not Bcl-2, until displaced by BH3-only proteins. Genes Dev 19: 1294–1305.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zamai L, Falcieri E, Zauli G, Cataldi A, Vitale M . (1993). Optimal detection of apoptosis by flow cytometry depends on cell morphology. Cytometry 14: 891–897.

    Article  CAS  PubMed  Google Scholar 

  • Zhang B, Gojo I, Fenton RG . (2002). Myeloid cell factor-1 is a critical survival factor for multiple myeloma. Blood 99: 1885–1893.

    Article  CAS  PubMed  Google Scholar 

  • Zhou P, Qian L, Bieszczad CK, Noelle R, Binder M, Levy NB et al. (1998). Mcl-1 in transgenic mice promotes survival in a spectrum of hematopoietic cell types and immortalization in the myeloid lineage. Blood 92: 3226–3239.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful for reagents provided by G Packham (University of Southampton School of Medicine, Southampton General Hospital, Southampton, UK). K Podar is the recipient of the Dunkin Donut's Rising Star Award 2007. This work was supported by a Multiple Myeloma Foundation (MMRF) Senior Research Grant Award (KP), National Institute of Health Grants RO CA 50947, PO-1 CA 78378, and P50 CA 10070; and the Doris Duke Distinguished Clinical Research Scientist Award (KCA).

Author information

Author notes

  1. K Podar and S L Gouill: These two authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medical Oncology, Jerome Lipper Multiple Myeloma Center, Harvard Medical School, Dana-Farber Cancer Institute, Boston, MA, USA

    K Podar, S L Gouill, J Zhang, Y-T Tai, T Hideshima, D Chauhan & K C Anderson

  2. INSERM U0601, Institut de biologie, 9 quai Moncousu and Service d'Hématologie clinique, Hôtel-Dieu CHU de Nantes, Nantes Cedex 01, France

    S L Gouill, M Amiot & J-L Harousseau

  3. Department of Biochemistry, St Jude Children's Research Hospital, Memphis, TN, USA

    J T Opferman

  4. Department of Medicine, Center for Hematologic Oncology, Dana-Farber Cancer Institute, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA

    E Zorn

Authors
  1. K Podar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S L Gouill
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J T Opferman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E Zorn
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Y-T Tai
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. T Hideshima
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. M Amiot
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. D Chauhan
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. J-L Harousseau
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. K C Anderson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to K Podar or K C Anderson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Podar, K., Gouill, S., Zhang, J. et al. A pivotal role for Mcl-1 in Bortezomib-induced apoptosis. Oncogene 27, 721–731 (2008). https://doi.org/10.1038/sj.onc.1210679

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1210679

Keywords

This article is cited by

Search

Advanced search

Quick links