Abstract
Antiapoptotic Bcl-2 family proteins are often highly expressed in chemotherapy-resistant cancers and impair mitochondrial outer membrane permeabilisation (MOMP), an important requirement for caspase activation via the intrinsic apoptosis pathway. Interestingly, although Bcl-2 overexpression in HeLa cervical cancer cells abrogated caspase processing in response to intrinsic apoptosis induction by staurosporine, tunicamycin or etoposide, residual caspase processing was observed following proteasome inhibition by bortezomib ([(1R)-3-methyl-1-({(2S)-3-phenyl-2-[(pyrazin-2-ylcarbonyl)amino]propanoyl}amino)butyl]boronic acid), epoxomicin (N-acetyl-N-methyl-lisoleucyl-L-isoleucyl-N-[(1S)-3-methyl-1-[[(2R)-2-methyloxiranyl]carbonyl]butyl]-L-threoninamide) or MG-132 (N-(benzyloxycarbonyl)leucinylleucinylleucinal). Similar responses were found in Bcl-2-overexpressing H460 NSCLC cells and Bax/Bak-deficient mouse embyronic fibroblasts. Mild caspase processing resulted in low DEVDase activities, which were MOMP independent and persisted for long periods without evoking immediate cell death. Surprisingly, depletion of caspase-3 and experiments in caspase-7-depleted MCF-7-Bcl-2 cells indicated that the DEVDase activity did not originate from effector caspases. Instead, Fas-associated death domain (FADD)-dependent caspase-8 activation was the major contributor to the slow, incomplete substrate cleavage. Caspase-8 activation was independent of death ligands, but required the induction of autophagy and the presence of Atg5. Depletion of XIAP or addition of XIAP-antagonising peptides resulted in a switch towards efficient apoptosis execution, suggesting that the requirement for MOMP was bypassed by activating the caspase-8/caspase-3 axis. Combination treatments of proteasome inhibitors and XIAP antagonists therefore represent a promising strategy to eliminate highly resistant cancer cells, which overexpress antiapoptotic Bcl-2 family members.
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Abbreviations
- 3-MA:
-
3-methyl adenine
- bortezomib:
-
[(1R)-3-methyl-1-({(2S)-3-phenyl-2-[(pyrazin-2-ylcarbonyl)amino]propanoyl}amino)butyl]boronic acid
- CFP:
-
cyan fluorescent protein
- CHX:
-
cycloheximide
- epoxomicin:
-
N-acetyl-N-methyl-lisoleucyl-L-isoleucyl-N-[(1S)-3-methyl-1-[[(2R)-2-methyloxiranyl]carbonyl]butyl]-L-threoninamide
- FasL:
-
fas ligand
- FRET:
-
Förster resonance energy transfer
- MG-132:
-
N-(benzyloxycarbonyl)leucinylleucinylleucinal
- MOMP:
-
mitochondrial outer membrane permeabilisation
- PI:
-
propidium iodide
- IMS-RP:
-
intermembrane space reporter protein
- STS:
-
staurosporine
- TMRM:
-
tetramethylrhodamine methylester
- TNF-α:
-
tumour necrosis factor-α
- TRAIL:
-
tumour necrosis factor-related apoptosis-inducing ligand
- YFP:
-
yellow fluorescent protein
- zVAD:
-
benzyloxycarbonyl-Val-Ala-Asp(O-methyl)-fluoromethylketone
References
Carlucci A, Lignitto L, Feliciello A . Control of mitochondria dynamics and oxidative metabolism by cAMP, AKAPs and the proteasome. Trends Cell Biol 2008; 18: 604–613.
Jung T, Catalgol B, Grune T . The proteasomal system. Mol Aspects Med 2009; 30: 191–296.
Brenner D, Mak TW . Mitochondrial cell death effectors. Curr Opin Cell Biol 2009; 21: 871–877.
Bhalla S, Balasubramanian S, David K, Sirisawad M, Buggy J, Mauro L et al. PCI-24781 induces caspase and reactive oxygen species-dependent apoptosis through NF-kappaB mechanisms and is synergistic with bortezomib in lymphoma cells. Clin Cancer Res 2009; 15: 3354–3365.
Colado E, Alvarez-Fernandez S, Maiso P, Martin-Sanchez J, Vidriales MB, Garayoa M et al. The effect of the proteasome inhibitor bortezomib on acute myeloid leukemia cells and drug resistance associated with the CD34+ immature phenotype. Haematologica 2008; 93: 57–66.
Li C, Li R, Grandis JR, Johnson DE . Bortezomib induces apoptosis via Bim and Bik up-regulation and synergizes with cisplatin in the killing of head and neck squamous cell carcinoma cells. Mol Cancer Ther 2008; 7: 1647–1655.
Tan TT, Degenhardt K, Nelson DA, Beaudoin B, Nieves-Neira W, Bouillet P et al. Key roles of BIM-driven apoptosis in epithelial tumors and rational chemotherapy. Cancer Cell 2005; 7: 227–238.
Nikrad M, Johnson T, Puthalalath H, Coultas L, Adams J, Kraft AS . The proteasome inhibitor bortezomib sensitizes cells to killing by death receptor ligand TRAIL via BH3-only proteins Bik and Bim. Mol Cancer Ther 2005; 4: 443–449.
Sayers TJ, Brooks AD, Koh CY, Ma W, Seki N, Raziuddin A et al. The proteasome inhibitor PS-341 sensitizes neoplastic cells to TRAIL-mediated apoptosis by reducing levels of c-FLIP. Blood 2003; 102: 303–310.
Ganten TM, Koschny R, Haas TL, Sykora J, Li-Weber M, Herzer K et al. Proteasome inhibition sensitizes hepatocellular carcinoma cells, but not human hepatocytes, to TRAIL. Hepatology 2005; 42: 588–597.
Hallett WH, Ames E, Motarjemi M, Barao I, Shanker A, Tamang DL et al. Sensitization of tumor cells to NK cell-mediated killing by proteasome inhibition. J Immunol 2008; 180: 163–170.
Galban S, Duckett CS . XIAP as a ubiquitin ligase in cellular signaling. Cell Death Differ 2010; 17: 54–60.
Vaux DL, Silke J . IAPs, RINGs and ubiquitylation. Nat Rev Mol Cell Biol 2005; 6: 287–297.
Rehm M, Dussmann H, Prehn JH . Real-time single cell analysis of Smac/DIABLO release during apoptosis. J Cell Biol 2003; 162: 1031–1043.
Fernandes-Alnemri T, Armstrong RC, Krebs J, Srinivasula SM, Wang L, Bullrich F et al. In vitro activation of CPP32 and Mch3 by Mch4, a novel human apoptotic cysteine protease containing two FADD-like domains. Proc Natl Acad Sci USA 1996; 93: 7464–7469.
Zhang XD, Gillespie SK, Hersey P . Staurosporine induces apoptosis of melanoma by both caspase-dependent and -independent apoptotic pathways. Mol Cancer Ther 2004; 3: 187–197.
Rehm M, Dussmann H, Janicke RU, Tavare JM, Kogel D, Prehn JH . Single-cell fluorescence resonance energy transfer analysis demonstrates that caspase activation during apoptosis is a rapid process. Role of caspase-3. J Biol Chem 2002; 277: 24506–24514.
McStay GP, Salvesen GS, Green DR . Overlapping cleavage motif selectivity of caspases: implications for analysis of apoptotic pathways. Cell Death Differ 2008; 15: 322–331.
Rehm M, Huber HJ, Dussmann H, Prehn JH . Systems analysis of effector caspase activation and its control by X-linked inhibitor of apoptosis protein. EMBO J 2006; 25: 4338–4349.
Hughes MA, Harper N, Butterworth M, Cain K, Cohen GM, MacFarlane M . Reconstitution of the death-inducing signaling complex reveals a substrate switch that determines CD95-mediated death or survival. Mol Cell 2009; 35: 265–279.
Pandey UB, Nie Z, Batlevi Y, McCray BA, Ritson GP, Nedelsky NB et al. HDAC6 rescues neurodegeneration and provides an essential link between autophagy and the UPS. Nature 2007; 447: 859–863.
Bell BD, Leverrier S, Weist BM, Newton RH, Arechiga AF, Luhrs KA et al. FADD and caspase-8 control the outcome of autophagic signaling in proliferating T cells. Proc Natl Acad Sci USA 2008; 105: 16677–16682.
Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, Outzen H et al. p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem 2007; 282: 24131–24145.
Yu L, Alva A, Su H, Dutt P, Freundt E, Welsh S et al. Regulation of an ATG7-beclin 1 program of autophagic cell death by caspase-8. Science 2004; 304: 1500–1502.
Ch’en IL, Beisner DR, Degterev A, Lynch C, Yuan J, Hoffmann A et al. Antigen-mediated T cell expansion regulated by parallel pathways of death. Proc Natl Acad Sci USA 2008; 105: 17463–17468.
Jost PJ, Grabow S, Gray D, McKenzie MD, Nachbur U, Huang DC et al. XIAP discriminates between type I and type II FAS-induced apoptosis. Nature 2009; 460: 1035–1039.
Fulda S, Wick W, Weller M, Debatin KM . Smac agonists sensitize for Apo2L/TRAIL- or anticancer drug-induced apoptosis and induce regression of malignant glioma in vivo. Nat Med 2002; 8: 808–815.
Miller CP, Ban K, Dujka ME, McConkey DJ, Munsell M, Palladino M et al. NPI-0052, a novel proteasome inhibitor, induces caspase-8 and ROS-dependent apoptosis alone and in combination with HDAC inhibitors in leukemia cells. Blood 2007; 110: 267–277.
Liu X, Yue P, Chen S, Hu L, Lonial S, Khuri FR et al. The proteasome inhibitor PS-341 (bortezomib) up-regulates DR5 expression leading to induction of apoptosis and enhancement of TRAIL-induced apoptosis despite up-regulation of c-FLIP and survivin expression in human NSCLC Cells. Cancer Res 2007; 67: 4981–4988.
Boatright KM, Renatus M, Scott FL, Sperandio S, Shin H, Pedersen IM et al. A unified model for apical caspase activation. Mol Cell 2003; 11: 529–541.
Arechiga AF, Bell BD, Leverrier S, Weist BM, Porter M, Wu Z et al. A Fas-associated death domain protein/caspase-8-signaling axis promotes S-phase entry and maintains S6 kinase activity in T cells responding to IL-2. J Immunol 2007; 179: 5291–5300.
Voortman J, Resende TP, Abou El Hassan MAI, Giaccone G, Kruyt FAE . TRAIL therapy in non-small cell lung cancer cells: sensitization to death receptor-mediated apoptosis by proteasome inhibitor bortezomib. Mol Cancer Therap 2007; 6: 2103–2112.
Caro-Maldonado A, Tait SW, Ramirez-Peinado S, Ricci JE, Fabregat I, Green DR et al. Glucose deprivation induces an atypical form of apoptosis mediated by caspase-8 in Bax-, Bak-deficient cells. Cell Death Differ 2010.
Pepper C, Lin TT, Pratt G, Hewamana S, Brennan P, Hiller L et al. Mcl-1 expression has in vitro and in vivo significance in chronic lymphocytic leukemia and is associated with other poor prognostic markers. Blood 2008; 112: 3807–3817.
Zhao WL, Daneshpouy ME, Mounier N, Briere J, Leboeuf C, Plassa LF et al. Prognostic significance of bcl-xL gene expression and apoptotic cell counts in follicular lymphoma. Blood 2004; 103: 695–697.
Yang Q, Moran MS, Haffty BG . Bcl-2 expression predicts local relapse for early-stage breast cancer receiving conserving surgery and radiotherapy. Breast Cancer Res Treat 2009; 115: 343–348.
Vogler M, Dinsdale D, Dyer MJ, Cohen GM . Bcl-2 inhibitors: small molecules with a big impact on cancer therapy. Cell Death Differ 2009; 16: 360–367.
Wilson TR, McEwan M, McLaughlin K, Le Clorennec C, Allen WL, Fennell DA et al. Combined inhibition of FLIP and XIAP induces Bax-independent apoptosis in type II colorectal cancer cells. Oncogene 2009; 28: 63–72.
Hellwig CT, Kohler BF, Lehtivarjo AK, Dussmann H, Courtney MJ, Prehn JH et al. Real time analysis of tumor necrosis factor-related apoptosis-inducing ligand/cycloheximide-induced caspase activities during apoptosis initiation. J Biol Chem 2008; 283: 21676–21685.
Flanagan L, Sebastia J, Tuffy LP, Spring A, Lichawska A, Devocelle M et al. XIAP impairs Smac release from the mitochondria during apoptosis. Cell Death and Dis 2010; 1: e49.
Acknowledgements
This work was supported by grants from the Research Committee of the Royal College of Surgeons in Ireland, the Health Research Board Ireland (RP/2006/258; RP/2008/7) and Science Foundation Ireland (09/RFP/BIC2375) awarded to MR, the National Biophotonics and Imaging Platform Ireland (Higher Education Authority PRTLI Cycle 4) and the European Union (FP7 Health – APO-SYS). We are grateful to Dr. Douglas Green (St. Jude Children's Research Hospital, Memphis), Dr. Frank Kruyt (University Medical Center Groningen), Drs. John Albeck and Peter Sorger (Harvard Medical School, Boston), Dr. Scott Oakes (University of California, San Francisco), Dr. Nathan Brady (German Cancer Research Center, Heidelberg), and Dr. Elaine Kay and Peter Jakob (Royal College of Surgeons in Ireland and Beaumont Hospital, Dublin) for the supply of cell lines, plasmids and additional materials. We also thank Eimear Costelloe, Kamil Jastrzebski and Dr. Tytus Bernas for technical assistance and helpful advice.
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Laussmann, M., Passante, E., Düssmann, H. et al. Proteasome inhibition can induce an autophagy-dependent apical activation of caspase-8. Cell Death Differ 18, 1584–1597 (2011). https://doi.org/10.1038/cdd.2011.27
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DOI: https://doi.org/10.1038/cdd.2011.27
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