Annals of Hematology

, Volume 91, Issue 6, pp 847–856

The proteasome inhibitor bortezomib targets cell cycle and apoptosis and acts synergistically in a sequence-dependent way with chemotherapeutic agents in mantle cell lymphoma

  • Grit Hutter
  • Malte Rieken
  • Alessandro Pastore
  • Oliver Weigert
  • Yvonne Zimmermann
  • Marc Weinkauf
  • Wolfgang Hiddemann
  • Martin Dreyling
Original Article


Single-agent bortezomib, a potent, selective, and reversible inhibitor of the 26S proteasome, has demonstrated clinical efficacy in relapsed and refractory mantle cell lymphoma (MCL). Objective response is achieved in up to 45% of the MCL patients; however, complete remission rates are low and duration of response proved to be relatively short. These limitations may be overcome by combining proteasome inhibition with conventional chemotherapy. Rational combination treatment and schedules require profound knowledge of underlying molecular mechanisms. Here we show that single-agent bortezomib treatment of MCL cell lines leads to G2/M arrest and induction of apoptosis accompanied by downregulation of EIF4E and CCND1 mRNA but upregulation of p15(INK4B) and p21 mRNA. We further present synergistic efficacy of bortezomib combined with cytarabine in MCL cell lines. Interestingly this sequence-dependent synergistic effect was seen almost exclusively in combination with AraC, indicating that pretreatment with cytarabine, followed by proteasome inhibition, may be the preferred approach.


MCL Bortezomib AraC Apoptosis Cell cycle arrest 


  1. 1.
    Ciechanover A (1994) The ubiquitin–proteasome proteolytic pathway. Cell 79:13–21PubMedCrossRefGoogle Scholar
  2. 2.
    Orlowski RZ, Kuhn DJ (2008) Proteasome inhibitors in cancer therapy: lessons from the first decade. Clin Cancer Res 14:1649–1657PubMedCrossRefGoogle Scholar
  3. 3.
    Adams J (2004) The development of proteasome inhibitors as anticancer drugs. Cancer Cells 5:417–421CrossRefGoogle Scholar
  4. 4.
    Montagut C, Rovira A, Albanell J (2006) The proteasome: a novel target for anticancer therapy. Clin Transl Oncol 8:313–317PubMedCrossRefGoogle Scholar
  5. 5.
    Richardson PG, Barlogie B, Berenson J et al (2003) A phase 2 study of bortezomib in relapsed, refractory myeloma. N Engl J Med 348:2609–2617PubMedCrossRefGoogle Scholar
  6. 6.
    Richardson PG, Mitsiades C, Ghobrial I, Anderson K (2006) Beyond single-agent bortezomib: combination regimens in relapsed multiple myeloma. Curr Opin Oncol 18:598–608PubMedCrossRefGoogle Scholar
  7. 7.
    Utecht KN, Kolesar J (2008) Bortezomib: a novel chemotherapeutic agent for hematologic malignancies. Am J Health Syst Pharm 65:1221–1231PubMedCrossRefGoogle Scholar
  8. 8.
    Gerecitano J, Portlock C, Moskowitz C et al (2009) Phase 2 study of weekly bortezomib in mantle cell and follicular lymphoma. Br J Haematol 146:652–655PubMedCrossRefGoogle Scholar
  9. 9.
    O’Connor OA, Moskowitz C, Portlock C et al (2009) Patients with chemotherapy-refractory mantle cell lymphoma experience high response rates and identical progression-free survivals compared with patients with relapsed disease following treatment with single agent bortezomib: results of a multicentre Phase 2 clinical trial. Br J Haematol 145:34–39PubMedCrossRefGoogle Scholar
  10. 10.
    Goy A, Bernstein SH, Kahl BS et al (2009) Bortezomib in patients with relapsed or refractory mantle cell lymphoma: updated time-to-event analyses of the multicenter phase 2 PINNACLE study. Ann Oncol 20:520–525PubMedCrossRefGoogle Scholar
  11. 11.
    Suh KS, Goy A (2008) Bortezomib in mantle cell lymphoma. Future Oncol 4:149–168PubMedCrossRefGoogle Scholar
  12. 12.
    Strauss SJ, Maharaj L, Hoare S et al (2006) Bortezomib therapy in patients with relapsed or refractory lymphoma: potential correlation of in vitro sensitivity and tumor necrosis factor alpha response with clinical activity. J Clin Oncol 24:2105–2112PubMedCrossRefGoogle Scholar
  13. 13.
    Fisher RI, Bernstein SH, Kahl BS et al (2006) Multicenter phase II study of bortezomib in patients with relapsed or refractory mantle cell lymphoma. J Clin Oncol 24:4867–4874PubMedCrossRefGoogle Scholar
  14. 14.
    Jeremias I, Debatin KM (1998) TRAIL induces apoptosis and activation of NFkappaB. Eur Cytokine Netw 9:687–688PubMedGoogle Scholar
  15. 15.
    Mitsiades N, Mitsiades CS, Richardson PG et al (2003) The proteasome inhibitor PS-341 potentiates sensitivity of multiple myeloma cells to conventional chemotherapeutic agents: therapeutic applications. Blood 101:2377–2380PubMedCrossRefGoogle Scholar
  16. 16.
    Duechler M, Linke A, Cebula B et al (2005) In vitro cytotoxic effect of proteasome inhibitor bortezomib in combination with purine nucleoside analogues on chronic lymphocytic leukaemia cells. Eur J Haematol 4:407–417CrossRefGoogle Scholar
  17. 17.
    Orlowski RZ (2005) The ubiquitin proteasome pathway from bench to bedside. Hematol Am Soc Hematol Educ Program 220–225Google Scholar
  18. 18.
    Aghajanian C, Soignet S, Dizon DS et al (2002) A phase I trial of the novel proteasome inhibitor PS341 in advanced solid tumor malignancies. Clin Cancer Res 8:2505–2511PubMedGoogle Scholar
  19. 19.
    Orlowski RZ, Stinchcombe TE, Mitchell BS et al (2002) Phase I trial of the proteasome inhibitor PS-341 in patients with refractory hematologic malignancies. J Clin Oncol 20:4420–4427PubMedCrossRefGoogle Scholar
  20. 20.
    Pérez-Galán P, Roué G, Villamor N, Campo E, Colomer D (2007) The BH3-mimetic GX15-070 synergizes with bortezomib in mantle cell lymphoma by enhancing Noxa-mediated activation of Bak. Blood 109:4441–4449PubMedCrossRefGoogle Scholar
  21. 21.
    Rolland D, Camara-Clayette V, Barbarat A et al (2008) Farnesyltransferase inhibitor R115777 inhibits cell growth and induces apoptosis in mantle cell lymphoma. Cancer Chemother Pharmacol 61:855–863PubMedCrossRefGoogle Scholar
  22. 22.
    Haritunians T, Mori A, O’Kelly J, Luong QT, Giles FJ, Koeffler HP (2007) Antiproliferative activity of RAD001 (everolimus) as a single agent and combined with other agents in mantle cell lymphoma. Leukemia 21:333–339PubMedCrossRefGoogle Scholar
  23. 23.
    Heider U, von Metzler I, Kaiser M et al (2008) Synergistic interaction of the histone deacetylase inhibitor SAHA with the proteasome inhibitor bortezomib in mantle cell lymphoma. Eur J Haematol 80:133–142PubMedCrossRefGoogle Scholar
  24. 24.
    Paoluzzi L, Gonen M, Bhagat G et al (2008) The BH3-only mimetic ABT-737 synergizes the antineoplastic activity of proteasome inhibitors in lymphoid malignancies. Blood 112:2906–2916PubMedCrossRefGoogle Scholar
  25. 25.
    Jones RJ, Chen Q, Voorhees PM et al (2008) Inhibition of the p53 E3 ligase HDM-2 induces apoptosis and DNA damage–independent p53 phosphorylation in mantle cell lymphoma. Clin Cancer Res 14:5416–5425PubMedCrossRefGoogle Scholar
  26. 26.
    Tabe Y, Sebasigari D, Jin L et al (2009) MDM2 antagonist nutlin-3 displays antiproliferative and proapoptotic activity in mantle cell lymphoma. Clin Cancer Res 15:933–942PubMedCrossRefGoogle Scholar
  27. 27.
    Rajkumar SV, Richardson PG, Hideshima T, Anderson KC (2005) Proteasome inhibition as a novel therapeutic target in human cancer. J Clin Oncol 23:630–639PubMedCrossRefGoogle Scholar
  28. 28.
    Chou TC (1991) The median-effect principle and the combination index for quantitation of synergism and antagonism. In: Chou TC, Rideout DC (eds) Synergism and antagonism in chemotherapy. Academic, San Diego, pp 61–102Google Scholar
  29. 29.
    Chou TC, Talalay P (1984) Quantitative analysis of doseeffect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 22:27–55PubMedCrossRefGoogle Scholar
  30. 30.
    Camps J, Salaverria I, Garcia MJ et al (2006) Genomic imbalances and patterns of karyotypic variability in mantle-cell lymphoma cell lines. Leuk Res 30:923–934PubMedCrossRefGoogle Scholar
  31. 31.
    Rosenwald A, Wright G, Wiestner A et al (2003) The proliferation gene expression signature is a quantitative integrator of oncogenic events that predicts survival in mantle cell lymphoma. Cancer Cells 3:185–197CrossRefGoogle Scholar
  32. 32.
    Bertoni F, Rinaldi A, Zucca E, Cavalli F (2006) Update on the molecular biology of mantle cell lymphoma. Hematol Oncol 24:22–27PubMedCrossRefGoogle Scholar
  33. 33.
    Bertoni F, Ponzoni M (2007) The cellular origin of mantle cell lymphoma. Int J Biochem Cell Biol 39:1747–1753PubMedCrossRefGoogle Scholar
  34. 34.
    Schmid M, Carson D (2000) Cell cycle regulation and hematological disorders. In: Beutler E, Lichtman MA (eds) 6th edn, McGraw-Hill Professional, pp 131–140Google Scholar
  35. 35.
    Muñoz-Alonso MJ, Acosta JC, Richard C, Delgado MD, Sedivy J, León J (2005) p21Cip1 and p27Kip1 Induce Distinct Cell Cycle Effects and Differentiation Programs in Myeloid Leukemia Cells. J Biol Chem 280:18120–18129PubMedCrossRefGoogle Scholar
  36. 36.
    Konicek BW, Dumstorf CA, Graff JR (2008) Targeting the eIF4F translation initiation complex for cancer therapy. Cell Cycle 15:2466–2471CrossRefGoogle Scholar
  37. 37.
    Gray NK, Wickens M (1998) Control of translation initiation in animals. Annu Rev Cell Dev Biol 14:399–458PubMedCrossRefGoogle Scholar
  38. 38.
    Larsson O, Li S, Issaenko OA, Avdulov S et al (2007) Eukaryotic translation initiation factor 4E induced progression of primary human mammary epithelial cells along the cancer pathway is associated with targeted translational deregulation of oncogenic drivers and inhibitors. Cancer Res 67:6814–6824PubMedCrossRefGoogle Scholar
  39. 39.
    Baiz D, Pozzato G, Dapas B et al (2009) Bortezomib arrests the proliferation of hepatocellular carcinoma cells HepG2 and JHH6 by differentially affecting E2F1, p21 and p27 levels. Biochimie 91:373–382PubMedCrossRefGoogle Scholar
  40. 40.
    Li Y, Dowbenko D, Lasky LA (2002) AKT/PKB phosphorylation of p21Cip/WAF1 enhances protein stability of p21Cip/WAF1 and promotes cell survival. J Biol Chem 277:11352–11361PubMedCrossRefGoogle Scholar
  41. 41.
    Ackler S, Ahmad S, Tobias C, Johnson MD, Glazer RI (2002) Delayed mammary gland involution in MMTV-AKT1 transgenic mice. Oncogene 21:198–206PubMedCrossRefGoogle Scholar
  42. 42.
    Li X, Liu J, Gao T (2009) beta-TrCP-mediated ubiquitination and degradation of PHLPP1 are negatively regulated by Akt. Mol Cell Biol 29:6192–6205PubMedCrossRefGoogle Scholar
  43. 43.
    Ma MH, Yang HH, Parker K et al (2003) The proteasome inhibitor PS-341 markedly enhances sensitivity of multiple myeloma tumor cells to chemotherapeutic agents. Clin Cancer Res 9:1136–1144PubMedGoogle Scholar
  44. 44.
    Dreyling, MH and Hiddemann W (2009) Current treatment standards and emerging strategies in mantle cell lymphoma. Hematology, ASH Education Program Book: 542–551Google Scholar
  45. 45.
    Sun XM, Butterworth M, MacFarlane M, Dubiel W, Ciechanover A, Cohen GM (2004) Caspase activation inhibits proteasome function during apoptosis. Mol Cell 14:81–93PubMedCrossRefGoogle Scholar
  46. 46.
    Pérez-Galán P, Roué 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 107:257–264PubMedCrossRefGoogle Scholar
  47. 47.
    Weigert O, Pastore A, Rieken M, Lang N, Hiddemann W, Dreyling M (2007) Sequence-dependent synergy of the proteasome inhibitor bortezomib and cytarabine in mantle cell lymphoma. Leukemia 21:524–528PubMedCrossRefGoogle Scholar
  48. 48.
    Weigert O, Weidmann E, Mueck R et al (2009) A novel regimen combining high dose cytarabine and bortezomib has activity in multiply relapsed and refractory mantle cell lymphoma—long-term results of a multicenter observation study. Leuk Lymphoma 50:716–722PubMedCrossRefGoogle Scholar
  49. 49.
    Congdon LM, Pourpak A, Escalante AM, Dorr RT, Landowski TH (2008) Proteasomal inhibition stabilizes topoisomerase IIalpha protein and reverses resistance to the topoisomerase II poison ethonafide (AMP-53, 6-ethoxyazonafide). Biochem Pharmacol 75(4):883–890PubMedCrossRefGoogle Scholar
  50. 50.
    Ma Y, Hendershot LM (2004) The role of the unfolded protein response in tumour development: friend or foe? Nat Rev Cancer 4(12):966–977PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Grit Hutter
    • 2
  • Malte Rieken
    • 3
  • Alessandro Pastore
    • 1
    • 2
  • Oliver Weigert
    • 1
    • 2
  • Yvonne Zimmermann
    • 2
  • Marc Weinkauf
    • 2
  • Wolfgang Hiddemann
    • 1
  • Martin Dreyling
    • 1
    • 2
  1. 1.Department of Medicine IIIUniversity Hospital GrosshadernMunichGermany
  2. 2.Helmholtz Centre MunichMuenchenGermany
  3. 3.Urologische KlinikUniversitätsspital BaselBaselSwitzerland

Personalised recommendations