Targeted proteasome inhibition by Velcade induces apoptosis in human mesothelioma and breast cancer cell lines



Thoracic malignancies and human breast cancer (HBC) continue to be aggressive solid tumors that are poor responders to the existing conventional standard chemotherapeutic approaches. Malignant pleural mesothelioma (MPM) is an asbestos-related tumor of the thoracic pleura that lacks effective treatment options. Altered ubiquitin proteasome pathway is frequently encountered in many malignancies including HBC and MPM and thus serves as an important target for therapeutic intervention strategies. Although proteasome inhibitor Velcade (Bortezomib) has been under clinical investigation for a number of cancers, limited preclinical studies with this agent have thus far been conducted in HBC and MPM malignancies.


To study the biological and molecular responses of MPM and HBC cells to Velcade treatments, and to identify mechanisms involved in transducing growth inhibitory effects of this agent.


Flow-cytometric analyses coupled with western immunoblotting and gene-array methodologies were utilized to determine mechanisms of Velcade-dependent growth suppression of five MPM (H2595, H2373, H2452, H2461, and H2714) and two breast cancer (MDA MB-468, SKBR-3) cell lines.


Our data revealed significant reduction in cell growth properties that were dose and time dependent. Velcade treatment resulted in G2M phase arrest, increased expression of cyclin-dependent kinase inhibitor p21 and pro-apoptotic protein Bax. Pretreatment of mesothelioma cells with Velcade showed synergistic effect with cisplatin combination regimens. High-throughput gene expression profiling among Velcade treated and untreated mesothelioma cell lines resulted in identification of novel transducers of apoptosis such as CARP-1, XAF1, and Troy proteins.


Velcade targets cell cycle and apoptosis signaling to suppress MPM and HBC growth in part by activating novel transducers of apoptosis. This pilot study has paved way for further in-depth analysis of the downstream target molecules associated with presensitization of mesothelioma cells in finding effective therapeutic treatment options for both mesothelioma and recalcitrant breast cancers.

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  1. 1.

    Antman KH, Pass HI, Schiff PB (2001) Benign and malignant mesothelioma. In: DeVita VTJ, Hellman S, Rosenberg SA (eds) Cancer: principles and practice of oncology, 6th edn. Lippincott-Raven, Philadelphia, pp 1943–1970

  2. 2.

    Peto J, Decaril A, La Vecchia C, Levi F, Negri E (1999) The European mesothelioma epidemic. Br J Cancer 79:666–672

    Article  CAS  PubMed  Google Scholar 

  3. 3.

    Sterman DH, Kaiser LR, Albelda SM (1999) Advances in the treatment of malignant pleural mesothelioma. Chest 116:504–520

    Article  CAS  PubMed  Google Scholar 

  4. 4.

    Ryan CW, Herndon J, Vogelzang NJ (1998) A review of chemotherapy trials for malignant mesothelioma. Chest 113:66S–73S

    Article  CAS  PubMed  Google Scholar 

  5. 5.

    Goudar RK (2005) New therapeutic options for mesothelioma. Curr Oncol Rep 7(4):260–265

    Article  CAS  PubMed  Google Scholar 

  6. 6.

    Krug LM (2005) An overview of chemotherapy for mesothelioma. Hematol Oncol Clin North Am 19(6):1117–1136

    Article  PubMed  Google Scholar 

  7. 7.

    Orlowski RZ, Dees EC (2003) Applying drugs that affect the ubiquitin-proteasome pathway to the therapy of breast cancer. Breast Cancer Res 5:1–7

    Article  CAS  PubMed  Google Scholar 

  8. 8.

    Richardson PG, Hideshima T, Anderson KC (2003) Bortezomib (PS-341): a novel, first-in-class proteasome inhibitor for the treatment of multiple myeloma and other cancers. Cancer Control 10(5):361–369 Review

    PubMed  Google Scholar 

  9. 9.

    Dou QP, Goldfarb RH (2002) Bortezomib/P341 (millennium pharmaceuticals) (invited review). IDrugs 5:828–834

    CAS  PubMed  Google Scholar 

  10. 10.

    Kane RC, Bross PF, Farrell AT, Pazdur R (2003) Velcade: U.S. FDA approval for the treatment of multiple myeloma progressing on prior therapy. Oncologist 8:508–513

    Article  PubMed  Google Scholar 

  11. 11.

    Adams J (2004) The proteosome: a suitable antineoplastic target. Nat Rev Cancer 4:349–360

    Article  CAS  PubMed  Google Scholar 

  12. 12.

    Rishi AK, Zhang L, Boyanapalli M, Wali A, Mohammad RM, Yu Y, Fontana JA, Hatfield JS, Dawson MI, Majumdar APN, Reichert U (2003) Identification and characterization of a Cell-Cycle and Apoptosis Regulatory Protein (CARP)-1 as a novel mediator of apoptosis signaling by Retinoid CD437. J Biol Chem 278:33422–33435

    Article  CAS  PubMed  Google Scholar 

  13. 13.

    Shao Z, Browning JL, Lee X, Scott ML, Shulga-Morskaya S, Allaire N, Thill G, Levesque M, Sah D, McCoy JM, Murray B, Jung V, Pepinsky RB, Mi S (2005) TAJ/TROY, an orphan TNF receptor family member, binds Nogo-66 receptor 1 and regulates axonal regeneration. Neuron 45:353–359

    Article  CAS  PubMed  Google Scholar 

  14. 14.

    Liston P, Fong WG, Kelly NL, Toji S, Miyazaki T, Conte D, Tamai K, Craig CG, McBurney MW, Korneluk RG (2001) Identification of XAF1 as an antagonist of XIAP anti-Caspase activity. Nat Cell Biol 3(2):128–133

    Article  CAS  PubMed  Google Scholar 

  15. 15.

    Pass HI, Lott D, Lonardo F, Harbut M, Liu Z, Tang N, Carbone M, Webb C, Wali A (2005) Asbestos exposure, pleural mesothelioma, and serum osteopontin levels. N Engl J Med 353(15):1564–1573

    Article  CAS  PubMed  Google Scholar 

  16. 16.

    Smyth GK, Limma L (2005) Liner models for microarray data. In: Gentleman R, Carey V, Duoit S, Irizarry R, Huber W (eds) Bioinformatics and computational biology solutions using R and bioconductor. Springer, New York, pp 397–420

    Google Scholar 

  17. 17.

    Tarca AL, Carey VJ, Chen XW, Romero R, Drãghici S (2007) Machine learning and its applications to biology. PLoS Comput Biol 3:e116

    Article  PubMed  Google Scholar 

  18. 18.

    Wali A, Morin PJ, Hough CD, Lonardo F, Seya T, Carbone M, Pass HI (2005) Identification of intelectin overexpression in malignant pleural mesothelioma by serial analysis of gene expression (SAGE). Lung Cancer 48(1):19–29

    Article  PubMed  Google Scholar 

  19. 19.

    Sartore-Bianchi A, Gasparri F, Galvani A, Nici L, Darnowski JW, Barbone D, Fennell DA, Gaudino G, Porta C, Mutti L (2007) Bortezomib inhibits nuclear factor-kappaB dependent survival and has potent in vivo activity in mesothelioma. Clin Cancer Res 13(19):5942–5951

    Article  CAS  PubMed  Google Scholar 

  20. 20.

    Lazebnik YA, Kaufmann SH, Desnoyers S, Poirier GG, Earnshaw WC (1994) Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature 371:346–347

    Article  CAS  PubMed  Google Scholar 

  21. 21.

    Rishi AK, Zhang L, Yu Y, Jiang Y, Nautiyal J, Wali A, Fontana JA, Levi E, Majumdar APN (2006) Cell cycle and apoptosis regulatory protein (CARP)-1 is involved in apoptosis signaling by epidermal growth factor receptor. J Biol Chem 281:13188–13198

    Article  CAS  PubMed  Google Scholar 

  22. 22.

    Kim JH, Yang CK, Heo K, Roeder RG, An W, Stallcup MR (2008) CCAR1, a key regulator of mediator complex recruitment to nuclear receptor transcription complexes. Mol Cell 31:510–519

    Article  CAS  PubMed  Google Scholar 

  23. 23.

    Ou CY, Kim JH, Yang CK, Stallcup MR (2009) Requirement of cell cycle and apoptosis regulator 1 for target gene activation by Wnt and {beta}-catenin and for anchorage-independent growth of human colon carcinoma cells. J Biol Chem 284(31):20629–20637

    Article  CAS  PubMed  Google Scholar 

  24. 24.

    Yuan BZ, Chapman JA, Reynolds SH (2008) Proteasome inhibitor MG132 induces apoptosis and inhibits invasion of human malignant pleural mesothelioma cells. Transl Oncol 1(3):129–140

    PubMed  Google Scholar 

  25. 25.

    Gordon GJ, Mani M, Mukhopadhyay L, Dong L, Yeap BY, Sugarbaker DJ, Bueno R (2007) Inhibitor of apoptosis proteins are regulated by tumour necrosis factor-alpha in malignant pleural mesothelioma. J Pathol 211(4):439–446

    Article  CAS  PubMed  Google Scholar 

  26. 26.

    Gordon GJ, Mani M, Maulik G, Mukhopadhyay L, Yeap BY, Kindler HL, Salgia R, Sugarbaker DJ, Bueno R (2008) Preclinical studies of the proteasome inhibitor bortezomib in malignant pleural mesothelioma. Cancer Chemother Pharmacol 61(4):549–558

    Article  CAS  PubMed  Google Scholar 

  27. 27.

    Borczuk AC, Cappellini GC, Kim HK, Hesdorffer M, Taub RN, Powell CA (2007) Molecular profiling of malignant peritoneal mesothelioma identifies the ubiquitin-proteasome pathway as a therapeutic target in poor prognosis tumors. Oncogene 26(4):610–617

    Article  CAS  PubMed  Google Scholar 

  28. 28.

    Fennell DA, Chacko A, Mutti L (2008) BCL-1 family regulation by the 20S proteasome inhibitor bortezomib. Oncogene 27:1189–1197

    Article  CAS  PubMed  Google Scholar 

  29. 29.

    Morohashi S, Kusumi T, Sato F, Odagiri H, Chiba H, Yoshihara S, Hakamada K, Sasaki M, Kijima H (2007) Decreased expression of claudin-1 correlates with recurrence status in breast cancer. Int J Mol Med 20:139–143

    PubMed  Google Scholar 

  30. 30.

    Tsukita S, Furuse M, Itoh M (2001) Multifunctional strands in tight junctions. Nat Rev Mol Cell Biol 2:285–293

    Article  CAS  PubMed  Google Scholar 

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This work was supported by the department of Veterans Affairs Merit Review grants to AKR and AW, Susan G. Komen for the Cure grant to AKR, and MARF support to AW.

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Correspondence to Arun K. Rishi.

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Wang, Y., Rishi, A.K., Puliyappadamba, V.T. et al. Targeted proteasome inhibition by Velcade induces apoptosis in human mesothelioma and breast cancer cell lines. Cancer Chemother Pharmacol 66, 455–466 (2010).

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  • Malignant pleural mesothelioma
  • Bortezomib (Velcade)
  • Apoptosis
  • Gene expression