Antioxidant Defenses Confer Resistance to High Dose Melphalan in Multiple Myeloma Cells
Abstract
:1. Introduction
2. Results
2.1. Antioxidants Protect Cells from Melphalan-Induced Toxicity
2.2. Addition of GSH Prevents Cell Cycle Arrest after Treatment with Melphalan and Increases Resistance to High Dose Melphalan
2.3. GSH Efficiently Blocks Melphalan Induced Cell Death in Primary Myeloma Cells of Patients
2.4. Glutathione Did Not Affect the Ability of Melphalan to Induce DNA Lesions in MM Cells
2.5. Melphalan Treatment Induces ROS Production, which are Involved in Melphalan-Induced Cytotoxicity and Prevented by Addition of GSH
2.6. Melphalan Induces GSH Depletion Together with Protein and Lipid Oxidation
2.7. Oxidative Stress Response Score is Associated with a Poor Outcome in MM Patients
3. Discussion
4. Materials and Methods
4.1. Drugs and Reagents
4.2. Human Myeloma Cell Lines Culture (HMCLs)
4.3. Primary Plasma Cells Culture
4.4. Cell Growth Assay
4.5. Cell Cycle Analysis
4.6. Caspase 3/7 and 9 Activity
4.7. Gene Expression Profiling and Statistical Analyses
4.8. DNA Repair Foci Quantification
4.9. Detection of Intracellular Reactive Oxygen Species (ROS)
4.10. Reduced Glutathione Quantification
4.11. Protein Carbonyl Oxidation Assay
4.12. Malondialdehyde (MDA) Quantification Assay
4.13. Study of Apoptosis
4.14. Statistical Analysis
4.15. Western Blot Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
- Ocio, E.M.; Richardson, P.G.; Rajkumar, S.V.; Palumbo, A.; Mateos, M.V.; Orlowski, R.; Kumar, S.; Usmani, S.; Roodman, D.; Niesvizky, R.; et al. New drugs and novel mechanisms of action in multiple myeloma in 2013: A report from the International Myeloma Working Group (IMWG). Leukemia 2014, 28, 525–542. [Google Scholar] [CrossRef] [PubMed]
- Van Rhee, F.; Giralt, S.; Barlogie, B. The future of autologous stem cell transplantation in myeloma. Blood 2014, 124, 328–333. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Spanswick, V.J.; Craddock, C.; Sekhar, M.; Mahendra, P.; Shankaranarayana, P.; Hughes, R.G.; Hochhauser, D.; Hartley, J.A. Repair of DNA interstrand crosslinks as a mechanism of clinical resistance to melphalan in multiple myeloma. Blood 2002, 100, 224–229. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Muniandy, P.A.; Liu, J.; Majumdar, A.; Liu, S.T.; Seidman, M.M. DNA interstrand crosslink repair in mammalian cells: Step by step. Crit. Rev. Biochem. Mol. Biol. 2010, 45, 23–49. [Google Scholar] [CrossRef] [PubMed]
- Deans, A.J.; West, S.C. DNA interstrand crosslink repair and cancer. Nat. Rev. Cancer 2011, 11, 467–480. [Google Scholar] [CrossRef] [Green Version]
- Gourzones-Dmitriev, C.; Kassambara, A.; Sahota, S.; Reme, T.; Moreaux, J.; Bourquard, P.; Hose, D.; Pasero, P.; Constantinou, A.; Klein, B. DNA repair pathways in human multiple myeloma: Role in oncogenesis and potential targets for treatment. Cell Cycle 2013, 12, 2760–2773. [Google Scholar] [CrossRef]
- Raymond, E.; Faivre, S.; Chaney, S.; Woynarowski, J.; Cvitkovic, E. Cellular and molecular pharmacology of oxaliplatin. Mol. Cancer Ther. 2002, 1, 227–235. [Google Scholar]
- Liu, H.; Lightfoot, R.; Stevens, J.L. Activation of heat shock factor by alkylating agents is triggered by glutathione depletion and oxidation of protein thiols. J. Biol. Chem. 1996, 271, 4805–4812. [Google Scholar]
- Spanswick, V.J.; Lowe, H.L.; Newton, C.; Bingham, J.P.; Bagnobianchi, A.; Kiakos, K.; Craddock, C.; Ledermann, J.A.; Hochhauser, D.; Hartley, J.A. Evidence for different mechanisms of ‘unhooking’ for melphalan and cisplatin-induced DNA interstrand cross-links in vitro and in clinical acquired resistant tumour samples. BMC Cancer 2012, 12, 436. [Google Scholar] [CrossRef]
- Hazlehurst, L.A.; Enkemann, S.A.; Beam, C.A.; Argilagos, R.F.; Painter, J.; Shain, K.H.; Saporta, S.; Boulware, D.; Moscinski, L.; Alsina, M.; et al. Genotypic and phenotypic comparisons of de novo and acquired melphalan resistance in an isogenic multiple myeloma cell line model. Cancer Res. 2003, 63, 7900–7906. [Google Scholar]
- Chen, Q.; Van der Sluis, P.C.; Boulware, D.; Hazlehurst, L.A.; Dalton, W.S. The FA/BRCA pathway is involved in melphalan-induced DNA interstrand cross-link repair and accounts for melphalan resistance in multiple myeloma cells. Blood 2005, 106, 698–705. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yang, Z.Z.; Chen, X.H.; Wang, D. Experimental study enhancing the chemosensitivity of multiple myeloma to melphalan by using a tissue-specific APE1-silencing RNA expression vector. Clin. Lymphoma Myeloma 2007, 7, 296–304. [Google Scholar] [CrossRef]
- Sousa, M.M.; Zub, K.A.; Aas, P.A.; Hanssen-Bauer, A.; Demirovic, A.; Sarno, A.; Tian, E.; Liabakk, N.B.; Slupphaug, G. An inverse switch in DNA base excision and strand break repair contributes to melphalan resistance in multiple myeloma cells. PLoS ONE 2013, 8, e55493. [Google Scholar] [CrossRef]
- Lin, J.; Raoof, D.A.; Thomas, D.G.; Greenson, J.K.; Giordano, T.J.; Robinson, G.S.; Bourner, M.J.; Bauer, C.T.; Orringer, M.B.; Beer, D.G. L-type amino acid transporter-1 overexpression and melphalan sensitivity in Barrett’s adenocarcinoma. Neoplasia 2004, 6, 74–84. [Google Scholar] [CrossRef]
- Kuhne, A.; Tzvetkov, M.V.; Hagos, Y.; Lage, H.; Burckhardt, G.; Brockmoller, J. Influx and efflux transport as determinants of melphalan cytotoxicity: Resistance to melphalan in MDR1 overexpressing tumor cell lines. Biochem. Pharmacol. 2009, 78, 45–53. [Google Scholar] [CrossRef] [PubMed]
- Viziteu, E.; Klein, B.; Basbous, J.; Lin, Y.L.; Hirtz, C.; Gourzones, C.; Tiers, L.; Bruyer, A.; Vincent, L.; Grandmougin, C.; et al. RECQ1 helicase is involved in replication stress survival and drug resistance in multiple myeloma. Leukemia 2017, 10, 2104–2113. [Google Scholar] [CrossRef]
- Trachootham, D.; Alexandre, J.; Huang, P. Targeting cancer cells by ROS-mediated mechanisms: A radical therapeutic approach? Nat. Rev. Drug Discov. 2009, 8, 579–591. [Google Scholar] [CrossRef] [PubMed]
- Brozovic, A.; Ambriovic-Ristov, A.; Osmak, M. The relationship between cisplatin-induced reactive oxygen species, glutathione, and BCL-2 and resistance to cisplatin. Crit. Rev. Toxicol. 2010, 40, 347–359. [Google Scholar] [CrossRef]
- Marullo, R.; Werner, E.; Degtyareva, N.; Moore, B.; Altavilla, G.; Ramalingam, S.S.; Doetsch, P.W. Cisplatin induces a mitochondrial-ROS response that contributes to cytotoxicity depending on mitochondrial redox status and bioenergetic functions. PLoS ONE 2013, 8, e81162. [Google Scholar] [CrossRef]
- Witte, A.B.; Anestal, K.; Jerremalm, E.; Ehrsson, H.; Arner, E.S. Inhibition of thioredoxin reductase but not of glutathione reductase by the major classes of alkylating and platinum-containing anticancer compounds. Free Radic. Biol. Med. 2005, 39, 696–703. [Google Scholar] [CrossRef]
- Surget, S.; Lemieux-Blanchard, E.; Maiga, S.; Descamps, G.; Le Gouill, S.; Moreau, P.; Amiot, M.; Pellat-Deceunynck, C. Bendamustine and melphalan kill myeloma cells similarly through reactive oxygen species production and activation of the p53 pathway and do not overcome resistance to each other. Leuk Lymphoma 2014, 55, 2165–2173. [Google Scholar] [CrossRef] [PubMed]
- Bellamy, W.T.; Dalton, W.S.; Gleason, M.C.; Grogan, T.M.; Trent, J.M. Development and characterization of a melphalan-resistant human multiple myeloma cell line. Cancer Res. 1991, 51, 995–1002. [Google Scholar]
- Moreaux, J.; Klein, B.; Bataille, R.; Descamps, G.; Maiga, S.; Hose, D.; Goldschmidt, H.; Jauch, A.; Reme, T.; Jourdan, M.; et al. A high-risk signature for patients with multiple myeloma established from the molecular classification of human myeloma cell lines. Haematologica 2011, 96, 574–582. [Google Scholar] [CrossRef] [PubMed]
- Hersh, M.R.; Ludden, T.M.; Kuhn, J.G.; Knight, W.A., 3rd. Pharmacokinetics of high dose melphalan. Investig. New Drugs 1983, 1, 331–334. [Google Scholar] [CrossRef]
- Traverso, N.; Ricciarelli, R.; Nitti, M.; Marengo, B.; Furfaro, A.L.; Pronzato, M.A.; Marinari, U.M.; Domenicotti, C. Role of glutathione in cancer progression and chemoresistance. Oxid. Med. Cell Longev. 2013, 2013, 972913. [Google Scholar] [CrossRef]
- Kucera, O.; Endlicher, R.; Rousar, T.; Lotkova, H.; Garnol, T.; Drahota, Z.; Cervinkova, Z. The effect of tert-butyl hydroperoxide-induced oxidative stress on lean and steatotic rat hepatocytes in vitro. Oxid. Med. Cell Longev. 2014, 2014, 752506. [Google Scholar] [CrossRef]
- Barrera, G. Oxidative stress and lipid peroxidation products in cancer progression and therapy. ISRN Oncol. 2012, 2012, 137289. [Google Scholar] [CrossRef]
- Wong, C.M.; Marcocci, L.; Liu, L.; Suzuki, Y.J. Cell signaling by protein carbonylation and decarbonylation. Antioxid. Redox Signal. 2010, 12, 393–404. [Google Scholar] [CrossRef]
- Wong, C.M.; Cheema, A.K.; Zhang, L.; Suzuki, Y.J. Protein carbonylation as a novel mechanism in redox signaling. Circ. Res. 2008, 102, 310–318. [Google Scholar] [CrossRef]
- Alam, J.; Stewart, D.; Touchard, C.; Boinapally, S.; Choi, A.M.; Cook, J.L. Nrf2, a Cap’n’Collar transcription factor, regulates induction of the heme oxygenase-1 gene. J. Biol. Chem. 1999, 274, 26071–26078. [Google Scholar] [CrossRef]
- Venugopal, R.; Jaiswal, A.K. Nrf1 and Nrf2 positively and c-Fos and Fra1 negatively regulate the human antioxidant response element-mediated expression of NAD(P)H:quinone oxidoreductase1 gene. Proc. Natl. Acad. Sci. USA 1996, 93, 14960–14965. [Google Scholar] [CrossRef] [Green Version]
- Solis, W.A.; Dalton, T.P.; Dieter, M.Z.; Freshwater, S.; Harrer, J.M.; He, L.; Shertzer, H.G.; Nebert, D.W. Glutamate-cysteine ligase modifier subunit: Mouse Gclm gene structure and regulation by agents that cause oxidative stress. Biochem. Pharmacol. 2002, 63, 1739–1754. [Google Scholar] [CrossRef]
- Moreaux, J.; Reme, T.; Leonard, W.; Veyrune, J.L.; Requirand, G.; Goldschmidt, H.; Hose, D.; Klein, B. Gene expression-based prediction of myeloma cell sensitivity to histone deacetylase inhibitors. Br. J. Cancer 2013, 109, 676–685. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Herviou, L.; Kassambara, A.; Boireau, S.; Robert, N.; Requirand, G.; Muller-Tidow, C.; Vincent, L.; Seckinger, A.; Goldschmidt, H.; Cartron, G.; et al. PRC2 targeting is a therapeutic strategy for EZ score defined high-risk multiple myeloma patients and overcome resistance to IMiDs. Clin. Epigenet. 2018, 10, 121. [Google Scholar] [CrossRef]
- Hothorn, T.; Lausen, B. On the exact distribution of maximally selected rank statistics. Comput. Stat. Data Anal. 2003, 43, 121–137. [Google Scholar] [CrossRef]
- Vikova, V.; Jourdan, M.; Robert, N.; Requirand, G.; Boireau, S.; Bruyer, A.; Vincent, L.; Cartron, G.; Klein, B.; Elemento, O.; et al. Comprehensive characterization of the mutational landscape in multiple myeloma cell lines reveals potential drivers and pathways associated with tumor progression and drug resistance. Theranostics 2019, 9, 540–553. [Google Scholar] [CrossRef] [PubMed]
- Anderson, K.C. The 39th David A. Karnofsky Lecture: Bench-to-bedside translation of targeted therapies in multiple myeloma. J. Clin. Oncol. 2012, 30, 445–452. [Google Scholar] [CrossRef]
- Paiva, B.; Gutierrez, N.C.; Rosinol, L.; Vidriales, M.B.; Montalban, M.A.; Martinez-Lopez, J.; Mateos, M.V.; Cibeira, M.T.; Cordon, L.; Oriol, A.; et al. High-risk cytogenetics and persistent minimal residual disease by multiparameter flow cytometry predict unsustained complete response after autologous stem cell transplantation in multiple myeloma. Blood 2012, 119, 687–691. [Google Scholar] [CrossRef]
- Rawstron, A.C.; Child, J.A.; de Tute, R.M.; Davies, F.E.; Gregory, W.M.; Bell, S.E.; Szubert, A.J.; Navarro-Coy, N.; Drayson, M.T.; Feyler, S.; et al. Minimal residual disease assessed by multiparameter flow cytometry in multiple myeloma: Impact on outcome in the Medical Research Council Myeloma IX Study. J. Clin. Oncol. 2013, 31, 2540–2547. [Google Scholar] [CrossRef] [PubMed]
- Paiva, B.; Vidriales, M.B.; Cervero, J.; Mateo, G.; Perez, J.J.; Montalban, M.A.; Sureda, A.; Montejano, L.; Gutierrez, N.C.; Garcia de Coca, A.; et al. Multiparameter flow cytometric remission is the most relevant prognostic factor for multiple myeloma patients who undergo autologous stem cell transplantation. Blood 2008, 112, 4017–4023. [Google Scholar] [CrossRef] [Green Version]
- Martinez-Lopez, J.; Lahuerta, J.J.; Pepin, F.; Gonzalez, M.; Barrio, S.; Ayala, R.; Puig, N.; Montalban, M.A.; Paiva, B.; Weng, L.; et al. Prognostic value of deep sequencing method for minimal residual disease detection in multiple myeloma. Blood 2014, 123, 3073–3079. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Rawstron, A.C.; Gregory, W.M.; de Tute, R.M.; Davies, F.E.; Bell, S.E.; Drayson, M.T.; Cook, G.; Jackson, G.H.; Morgan, G.J.; Child, J.A.; et al. Minimal residual disease in myeloma by flow cytometry: Independent prediction of survival benefit per log reduction. Blood 2015, 125, 1932–1935. [Google Scholar] [CrossRef] [PubMed]
- Caraux, A.; Vincent, L.; Bouhya, S.; Quittet, P.; Moreaux, J.; Requirand, G.; Veyrune, J.L.; Olivier, G.; Cartron, G.; Rossi, J.F.; et al. Residual malignant and normal plasma cells shortly after high dose melphalan and stem cell transplantation. Highlight of a putative therapeutic window in Multiple Myeloma? Oncotarget 2012, 3, 1335–1347. [Google Scholar] [CrossRef] [Green Version]
- Ling, Y.H.; Liebes, L.; Zou, Y.; Perez-Soler, R. Reactive oxygen species generation and mitochondrial dysfunction in the apoptotic response to Bortezomib, a novel proteasome inhibitor, in human H460 non-small cell lung cancer cells. J. Biol. Chem. 2003, 278, 33714–33723. [Google Scholar] [CrossRef] [PubMed]
- Boldogh, I.; Roy, G.; Lee, M.S.; Bacsi, A.; Hazra, T.K.; Bhakat, K.K.; Das, G.C.; Mitra, S. Reduced DNA double strand breaks in chlorambucil resistant cells are related to high DNA-PKcs activity and low oxidative stress. Toxicology 2003, 193, 137–152. [Google Scholar] [CrossRef] [PubMed]
- Dusinska, M.; Staruchova, M.; Horska, A.; Smolkova, B.; Collins, A.; Bonassi, S.; Volkovova, K. Are glutathione S transferases involved in DNA damage signalling? Interactions with DNA damage and repair revealed from molecular epidemiology studies. Mutat. Res. 2012, 736, 130–137. [Google Scholar] [CrossRef] [PubMed]
- Pujari, G.; Berni, A.; Palitti, F.; Chatterjee, A. Influence of glutathione levels on radiation-induced chromosomal DNA damage and repair in human peripheral lymphocytes. Mutat. Res. 2009, 675, 23–28. [Google Scholar] [CrossRef]
- Franco, R.; Cidlowski, J.A. Apoptosis and glutathione: Beyond an antioxidant. Cell Death Differ. 2009, 16, 1303–1314. [Google Scholar] [CrossRef] [PubMed]
- Niture, S.K.; Jaiswal, A.K. Nrf2 protein up-regulates antiapoptotic protein Bcl-2 and prevents cellular apoptosis. J. Biol. Chem. 2012, 287, 9873–9886. [Google Scholar] [CrossRef] [PubMed]
- Hour, T.C.; Huang, C.Y.; Lin, C.C.; Chen, J.; Guan, J.Y.; Lee, J.M.; Pu, Y.S. Characterization of molecular events in a series of bladder urothelial carcinoma cell lines with progressive resistance to arsenic trioxide. Anticancer Drugs 2004, 15, 779–785. [Google Scholar] [CrossRef]
- Tafani, M.; Sansone, L.; Limana, F.; Arcangeli, T.; De Santis, E.; Polese, M.; Fini, M.; Russo, M.A. The Interplay of Reactive Oxygen Species, Hypoxia, Inflammation, and Sirtuins in Cancer Initiation and Progression. Oxid. Med. Cell Longev. 2016, 2016, 3907147. [Google Scholar] [CrossRef]
- Zanotto-Filho, A.; Masamsetti, V.P.; Loranc, E.; Tonapi, S.S.; Gorthi, A.; Bernard, X.; Goncalves, R.M.; Moreira, J.C.; Chen, Y.; Bishop, A.J. Alkylating Agent-Induced NRF2 Blocks Endoplasmic Reticulum Stress-Mediated Apoptosis via Control of Glutathione Pools and Protein Thiol Homeostasis. Mol. Cancer Ther. 2016, 15, 3000–3014. [Google Scholar] [CrossRef]
- Wallington-Beddoe, C.T.; Pitson, S.M. Enhancing ER stress in myeloma. Aging 2017, 9, 1645–1646. [Google Scholar] [CrossRef] [Green Version]
- Nicolussi, A.; D’Inzeo, S.; Capalbo, C.; Giannini, G.; Coppa, A. The role of peroxiredoxins in cancer. Mol. Clin. Oncol. 2017, 6, 139–153. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Papa, L.; Manfredi, G.; Germain, D. SOD1, an unexpected novel target for cancer therapy. Genes Cancer 2014, 5, 15–21. [Google Scholar] [CrossRef] [Green Version]
- Marchesini, M.; Ogoti, Y.; Fiorini, E.; Aktas Samur, A.; Nezi, L.; D’Anca, M.; Storti, P.; Samur, M.K.; Ganan-Gomez, I.; Fulciniti, M.T.; et al. ILF2 Is a Regulator of RNA Splicing and DNA Damage Response in 1q21-Amplified Multiple Myeloma. Cancer Cell 2017, 32, 88–100. [Google Scholar] [CrossRef] [PubMed]
- Moreaux, J.; Hose, D.; Jourdan, M.; Reme, T.; Hundemer, M.; Moos, M.; Robert, N.; Moine, P.; De Vos, J.; Goldschmidt, H.; et al. TACI expression is associated with a mature bone marrow plasma cell signature and C-MAF overexpression in human myeloma cell lines. Haematologica 2007, 92, 803–811. [Google Scholar] [CrossRef]
- Moreaux, J.; Reme, T.; Leonard, W.; Veyrune, J.L.; Requirand, G.; Goldschmidt, H.; Hose, D.; Klein, B. Development of gene expression-based score to predict sensitivity of multiple myeloma cells to DNA methylation inhibitors. Mol. Cancer Ther. 2012, 11, 2685–2692. [Google Scholar] [CrossRef] [PubMed]
- Kassambara, A.; Reme, T.; Jourdan, M.; Fest, T.; Hose, D.; Tarte, K.; Klein, B. GenomicScape: An easy-to-use web tool for gene expression data analysis. Application to investigate the molecular events in the differentiation of B cells into plasma cells. PLoS Comput. Biol. 2015, 11, e1004077. [Google Scholar] [CrossRef]
- Gentleman, R.C.; Carey, V.J.; Bates, D.M.; Bolstad, B.; Dettling, M.; Dudoit, S.; Ellis, B.; Gautier, L.; Ge, Y.; Gentry, J.; et al. Bioconductor: Open software development for computational biology and bioinformatics. Genome Biol. 2004, 5, R80. [Google Scholar] [CrossRef]
- Kitamura, H.; Motohashi, H. NRF2 addiction in cancer cells. Cancer Sci. 2018, 109, 900–911. [Google Scholar] [CrossRef] [PubMed]
- Rojo de la Vega, M.; Dodson, M.; Chapman, E.; Zhang, D.D. NRF2-targeted therapeutics: New targets and modes of NRF2 regulation. Curr. Opin. Toxicol. 2016, 1, 62–70. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gorrini, C.; Harris, I.S.; Mak, T.W. Modulation of oxidative stress as an anticancer strategy. Nat. Rev. Drug Discov. 2013, 12, 931–947. [Google Scholar] [CrossRef] [PubMed]
© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Gourzones, C.; Bellanger, C.; Lamure, S.; Gadacha, O.K.; De Paco, E.G.; Vincent, L.; Cartron, G.; Klein, B.; Moreaux, J. Antioxidant Defenses Confer Resistance to High Dose Melphalan in Multiple Myeloma Cells. Cancers 2019, 11, 439. https://doi.org/10.3390/cancers11040439
Gourzones C, Bellanger C, Lamure S, Gadacha OK, De Paco EG, Vincent L, Cartron G, Klein B, Moreaux J. Antioxidant Defenses Confer Resistance to High Dose Melphalan in Multiple Myeloma Cells. Cancers. 2019; 11(4):439. https://doi.org/10.3390/cancers11040439
Chicago/Turabian StyleGourzones, Claire, Céline Bellanger, Sylvain Lamure, Ouissem Karmous Gadacha, Elvira Garcia De Paco, Laure Vincent, Guillaume Cartron, Bernard Klein, and Jérôme Moreaux. 2019. "Antioxidant Defenses Confer Resistance to High Dose Melphalan in Multiple Myeloma Cells" Cancers 11, no. 4: 439. https://doi.org/10.3390/cancers11040439