Abstract
The proteasome has emerged as a novel target for antineoplastic treatment of hematological malignancies and solid tumors, including those of the central nervous system. To identify cell death pathways activated in response to inhibition of the proteasome system in cancer cells, we treated human SH-SY5Y neuroblastoma cells with the selective proteasome inhibitor (PI) epoxomicin (Epoxo). Prolonged exposure to Epoxo was associated with increased levels of poly-ubiquitinylated proteins and p53, release of cytochrome c from the mitochondria, and activation of caspases. Analysis of global gene expression using high-density oligonucleotide microarrays revealed that Epoxo triggered transcriptional activation of the two Bcl-2-homology domain-3-only (BH3-only) genes p53 upregulated modulator of apoptosis (PUMA) and Bim. Subsequent studies in PUMA- and Bim-deficient cells indicated that Epoxo-induced caspase activation and apoptosis was predominantly PUMA-dependent. Further characterization of the transcriptional response to Epoxo in HCT116 human colon cancer cells demonstrated that PUMA induction was p53-dependent; with deficiency in either p53 or PUMA significantly protected HCT116 cells against Epoxo-induced apoptosis. Our data suggest that p53 activation and the transcriptional induction of its target gene PUMA play an important role in the sensitivity of cancer cells to apoptosis induced by proteasome inhibition, and imply that antineoplastic therapies with PIs might be especially useful in cancers with functional p53.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
Accessions
GenBank/EMBL/DDBJ
Abbreviations
- PUMA:
-
p53 upregulated modulator of apoptosis
- GRP78 :
-
immunoglobulin heavy chain-binding protein/78 kDa glucose-regulated protein
- DMSO:
-
dimethylsulfoxide
- CHOP :
-
C/EBP-homologous protein
- Epoxo:
-
epoxomicin
- WT:
-
wild type
References
Adams J . (2002). Proteasome inhibitors as new anticancer drugs. Curr Opin Oncol 14: 628–634.
Adams J . (2004a). The proteasome: a suitable antineoplastic target. Nat Rev Cancer 4: 349.
Adams J . (2004b). The proteasome: a suitable antineoplastic target. Nat Rev Cancer 4: 349–360.
Aghajanian C, Soignet S, Dizon DS, Pien CS, Adams J, Elliott PJ et al. (2002). A phase I trial of the novel proteasome inhibitor PS341 in advanced solid tumor malignancies. Clin Cancer Res 8: 2505–2511.
Amiri KI, Horton LW, LaFleur BJ, Sosman JA, Richmond A . (2004). Augmenting chemosensitivity of malignant melanoma tumors via proteasome inhibition: implication for bortezomib (VELCADE, PS-341) as a therapeutic agent for malignant melanoma. Cancer Res 64: 4912–4918.
Anan A, Baskin-Bey ES, Bronk SF, Werneburg NW, Shah VH, Gores GJ . (2006). Proteasome inhibition induces hepatic stellate cell apoptosis. Hepatology 43: 335–344.
Biswas SC, Ryu E, Park C, Malagelada C, Greene LA . (2005). Puma and p53 play required roles in death evoked in a cellular model of Parkinson disease. Neurochem Res 30: 839–845.
Bouillet P, Metcalf D, Huang DC, Tarlinton DM, Kay TW, Kontgen F et al. (1999). Proapoptotic Bcl-2 relative Bim required for certain apoptotic responses, leukocyte homeostasis, and to preclude autoimmunity. Science 286: 1735–1738.
Breitschopf K, Zeiher AM, Dimmeler S . (2000). Ubiquitin-mediated degradation of the proapoptotic active form of bid. A functional consequence on apoptosis induction. J Biol Chem 275: 21648–21652.
Ceballos E, Munoz-Alonso MJ, Berwanger B, Acosta JC, Hernandez R, Krause M et al. (2005). Inhibitory effect of c-Myc on p53-induced apoptosis in leukemia cells. Microarray analysis reveals defective induction of p53 target genes and upregulation of chaperone genes. Oncogene 24: 4559–4571.
Chauhan D, Hideshima T, Anderson KC . (2005). Proteasome inhibition in multiple myeloma: therapeutic implication. Annu Rev Pharmacol Toxicol 45: 465–476.
Chowdary DR, Dermody JJ, Jha KK, Ozer HL . (1994). Accumulation of p53 in a mutant cell line defective in the ubiquitin pathway. Mol Cell Biol 14: 1997–2003.
Dang CV . (1999). c-Myc target genes involved in cell growth, apoptosis, and metabolism. Mol Cell Biol 19: 1–11.
Ganten TM, Koschny R, Haas TL, Sykora J, Li-Weber M, Herzer K et al. (2005). Proteasome inhibition sensitizes hepatocellular carcinoma cells, but not human hepatocytes, to TRAIL. Hepatology 42: 588–597.
Han J-w, Flemington C, Houghton AB, Gu Z, Zambetti GP, Lutz RJ et al. (2001). Expression of bbc3, a pro-apoptotic BH3-only gene, is regulated by diverse cell death and survival signals. Proc Natl Acad Sci USA 98: 11318–11323.
Hanada M, Sugawara K, Kaneta K, Toda S, Nishiyama Y, Tomita K et al. (1992). Epoxomicin, a new antitumor agent of microbial origin. J Antibiotics, 1746–1752.
Hideshima T, Chauhan D, Richardson P, Mitsiades C, Mitsiades N, Hayashi T et al. (2002). NF-kappa B as a therapeutic target in multiple myeloma. J Biol Chem 277: 16639–16647.
Huang H-k, Joazeiro CAP, Bonfoco E, Kamada S, Leverson JD, Hunter T . (2000). The inhibitor of apoptosis, cIAP2, functions as a ubiquitin-protein ligase and promotes in vitro monoubiquitination of caspases 3 and 7. J Biol Chem 275: 26661–26664.
Karin M . (2006). Nuclear factor-[kappa]B in cancer development and progression. Nature 441: 431.
Karin M, Ben-Neriah Y . (2000). Phosphorylation meets ubiquitination: the control of NF-[kappa]B activity. Annu Rev Immunol 18: 621–663.
Kikuchi S, Shinpo K, Tsuji S, Takeuchi M, Yamagishi S, Makita Z et al. (2003). Effect of proteasome inhibitor on cultured mesencephalic dopaminergic neurons. Brain Res 964: 228.
Kurland JF, Meyn RE . (2001). Protease inhibitors restore radiation-induced apoptosis to Bcl-2-expressing lymphoma cells. Int J Cancer 96: 327–333.
Kuwana T, Bouchier-Hayes L, Chipuk JE, Bonzon C, Sullivan BA, Green DR et al. (2005). BH3 domains of BH3-only proteins differentially regulate Bax-mediated mitochondrial membrane permeabilization both directly and indirectly. Mol Cell 17: 525–535.
Leverkus M, Sprick MR, Wachter T, Mengling T, Baumann B, Serfling E et al. (2003). 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.
Ley R, Balmanno K, Hadfield K, Weston C, Cook SJ . (2003). Activation of the ERK1/2 signaling pathway promotes phosphorylation and proteasome-dependent degradation of the BH3-only protein, Bim. J Biol Chem 278: 18811–18816.
Luo JL, Kamata H, Karin M . (2005). IKK/NF-kappaB signaling: balancing life and death – a new approach to cancer therapy. J Clin Invest 115: 2625–2632.
Marshansky V, Wang X, Bertrand R, Luo H, Duguid W, Chinnadurai G et al. (2001). Proteasomes modulate balance among proapoptotic and antiapoptotic Bcl-2 family members and compromise functioning of the electron transport chain in leukemic cells. J Immunol 166: 3130–3142.
Masdehors P, Merle-Beral H, Maloum K, Omura S, Magdelenat H, Delic J . (2000). Deregulation of the ubiquitin system and p53 proteolysis modify the apoptotic response in B-CLL lymphocytes. Blood 96: 269–274.
Meiners S, Heyken D, Weller A, Ludwig A, Stangl K, Kloetzel PM et al. (2003). Inhibition of proteasome activity induces concerted expression of proteasome genes and de novo formation of Mammalian proteasomes. J Biol Chem 278: 21517–21525.
Melino G . (2005). Discovery of the ubiquitin proteasome system and its involvement in apoptosis. 12: 1155.
Meng L, Mohan R, Kwok BH, Elofsson M, Sin N, Crews CM . (1999). Epoxomicin, a potent and selective proteasome inhibitor, exhibits in vivo antiinflammatory activity. Proc Natl Acad Sci USA 96: 10403–10408.
Mitsiades N, Mitsiades CS, Poulaki V, Chauhan D, Fanourakis G, Gu X et al. (2002). Molecular sequelae of proteasome inhibition in human multiple myeloma cells. 99: 14374–14379.
Nakano K, Vousden KH . (2001). PUMA, a novel proapoptotic gene, is induced by p53. Mol Cell 7: 683–694.
Nikrad M, Johnson T, Puthalalath H, Coultas L, Adams J, Kraft AS . (2005). 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.
Owen-Schaub LB, Zhang W, Cusack JC, Angelo LS, Santee SM, Fujiwara T et al. (1995). Wild-type human p53 and a temperature-sensitive mutant induce Fas/APO-1 expression. Mol Cell Biol 15: 3032–3040.
Palombella VJ, Rando OJ, Goldberg AL, Maniatis T . (1994). The ubiquitinproteasome pathway is required for processing the NF-[kappa]B1 precursor protein and the activation of NF-[kappa]B. Cell 78: 773.
Papandreou CN, Daliani DD, Nix D, Yang H, Madden T, Wang X et al. (2004). Phase I trial of the proteasome inhibitor bortezomib in patients with advanced solid tumors with observations in androgen-independent prostate cancer. J Clin Oncol 22: 2108–2121.
Park DJ, Lenz HJ . (2004). The role of proteasome inhibitors in solid tumors. Ann Med 36: 296–303.
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.
Reimertz C, Kogel D, Lankiewicz S, Poppe M, Prehn JH . (2001). Ca(2+)-induced inhibition of apoptosis in human SH-SY5Y neuroblastoma cells: degradation of apoptotic protease activating factor-1 (APAF-1). J Neurochem 78: 1256–1266.
Reimertz C, Kogel D, Rami A, Chittenden T, Prehn JH . (2003). Gene expression during ER stress-induced apoptosis in neurons: induction of the BH3-only protein Bbc3/PUMA and activation of the mitochondrial apoptosis pathway. J Cell Biol 162: 587–597.
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.
Shinohara K, Tomioka M, Nakano H, Tone S, Ito H, Kawashima S . (1996). Apoptosis induction resulting from proteasome inhibition. Biochem J 317 (Part 2): 385–388.
Suzuki Y, Nakabayashi Y, Takahashi R . (2001). Ubiquitin-protein ligase activity of X-linked inhibitor of apoptosis protein promotes proteasomal degradation of caspase-3 and enhances its anti-apoptotic effect in Fas-induced cell death. Proc Natl Acad Sci USA 98: 8662–8667.
Tan T-T, Degenhardt K, Nelson DA, Beaudoin B, Nieves-Neira W, Bouillet P et al. (2005). Key roles of BIM-driven apoptosis in epithelial tumors and rational chemotherapy. Cancer Cell 7: 227.
Villunger A, Michalak EM, Coultas L, Mullauer F, Bock G, Ausserlechner MJ et al. (2003a). p53- and drug-induced apoptotic responses mediated by BH3-only proteins Puma and Noxa. Science 302: 1036–1038.
Villunger A, Scott C, Bouillet P, Strasser A . (2003b). Essential role for the BH3-only protein Bim but redundant roles for Bax, Bcl-2, and Bcl-w in the control of granulocyte survival. Blood 101: 2393–2400.
Wagenknecht B, Hermisson M, Eitel K, Weller M . (1999). Proteasome inhibitors induce p53/p21-independent apoptosis in human glioma cells. Cell Physiol Biochem 9: 117–125.
Wagenknecht B, Hermisson M, Groscurth P, Liston P, Krammer PH, Weller M . (2000). Proteasome inhibitor-induced apoptosis of glioma cells involves the processing of multiple caspases and cytochrome c release. J Neurochem 75: 2288–2297.
Willis SN, Adams JM . (2005). Life in the balance: how BH3-only proteins induce apoptosis. Curr Opin Cell Biol 17: 617.
Wong HK, Fricker M, Wyttenbach A, Villunger A, Michalak EM, Strasser A et al. (2005). Mutually exclusive subsets of BH3-only proteins are activated by the p53 and c-Jun N-terminal kinase/c-Jun signaling pathways during cortical neuron apoptosis induced by arsenite. Mol Cell Biol 25: 8732–8747.
Wu GS, Burns TF, McDonald III ER, Jiang W, Meng R, Krantz ID et al. (1997). KILLER/DR5 is a DNA damage-inducible p53-regulated death receptor gene. Nat Genet 17: 141–143.
Yan C, Lu D, Hai T, Boyd DD . (2005). Activating transcription factor 3, a stress sensor, activates p53 by blocking its ubiquitination. EMBO J 24: 2425–2435.
Yin D, Zhou H, Kumagai T, Liu G, Ong JM, Black KL et al. (2005). Proteasome inhibitor PS-341 causes cell growth arrest and apoptosis in human glioblastoma multiforme (GBM). Oncogene 24: 344–354.
Yu J, Wang Z, Kinzler KW, Vogelstein B, Zhang L . (2003). PUMA mediates the apoptotic response to p53 in colorectal cancer cells. Proc Natl Acad Sci USA 100: 1931–1936.
Yu J, Zhang L, Hwang PM, Kinzler KW, Vogelstein B . (2001). PUMA induces the rapid apoptosis of colorectal cancer cells. Mol Cell 7: 673–682.
Acknowledgements
The authors wish to thank Drs B Vogelstein, L Zhang, and J Yu for p53 and PUMA-deficient cells and Drs P Bouillet and J Adams for gifts of knockout mice. This study was supported by grants from the DFG (PR 338/9-3 and 9-4) to JHMP and DK and Science Foundation Ireland (03/RP/B344) to JHMP.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).
Rights and permissions
About this article
Cite this article
Concannon, C., Koehler, B., Reimertz, C. et al. Apoptosis induced by proteasome inhibition in cancer cells: predominant role of the p53/PUMA pathway. Oncogene 26, 1681–1692 (2007). https://doi.org/10.1038/sj.onc.1209974
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1209974
Keywords
This article is cited by
-
Inhibition of noncaspase proteases, calpain and proteasome, via ALLN and Bortezomib contributes to cell death through low degradation of pro-/anti-apoptotic proteins and apoptosis induction
Medical Oncology (2022)
-
A new combination strategy to enhance apoptosis in cancer cells by using nanoparticles as biocompatible drug delivery carriers
Scientific Reports (2021)
-
Marizomib sensitizes primary glioma cells to apoptosis induced by a latest-generation TRAIL receptor agonist
Cell Death & Disease (2021)
-
Bortezomib inhibits growth and sensitizes glioma to temozolomide (TMZ) via down-regulating the FOXM1–Survivin axis
Cancer Communications (2019)
-
Anti-cancer activities of cytokinin ribosides
Phytochemistry Reviews (2019)