Abstract
Heat shock protein 70 (HSP70), a chaperone protein associated with tumorigenesis and chemoresistance, has attracted significant attention as a potential therapeutic target for the development of anticancer drugs. Here, the effects of pifithrin-μ, an effective dual inhibitor of HSP70 and p53, on anticancer activities and epithelial–mesenchymal transition (EMT) were investigated in malignant mesothelioma (MM) cells. MSTO-211HAcT cells, pre-incubated in a medium containing lactic acid, showed more potent resistance to cisplatin and gemcitabine, compared with their acid-sensitive parental MSTO-211H cells. Pifithrin-μ treatment induced both apoptosis and necroptosis, which were accompanied by an EMT-like phenomenon, as evidenced by an elongated cell morphology, decreased levels of epithelial cell markers including E-cadherin, claudin-1, and β-catenin, increased levels of mesenchymal markers including Snail, Slug, and vimentin, and increased cell migratory property. Moreover, pifithrin-μ increased intracellular ROS levels, which is associated with mitochondrial dysfunction and decreased cellular ATP content. A series of changes caused by pifithrin-μ treatment were effectively restored by lowering the ROS level through pretreatment with N-acetylcysteine. Collectively, our results suggest that pifithrin-μ may promote the metastatic behavior of surviving cells by triggering the EMT, despite its effective cell-killing action against MM cells, possibly linked to oxidative mitochondrial dysfunction and ATP depletion.
References
Altomare DA, Menges CW, Xu J, Pei J, Zhang L, Tadevosyan A, Neumann-Domer E, Liu Z, Carbone M, Chudoba I, Klein-Szanto AJ, Testa JR (2011) Losses of both products of the Cdkn2a/Arf locus contribute to asbestos-induced mesothelioma development and cooperate to accelerate tumorigenesis. PLoS ONE 6:e18828
Chang CJ, Chao CH, Xia W, Yang JY, Xiong Y, Li CW, Yu WH, Rehman SK, Hsu JL, Lee HH, Liu M, Chen CT, Yu D, Hung MC (2011) p53 regulates epithelial–mesenchymal transition and stem cell properties through modulating miRNAs. Nat Cell Biol 13:317–323
Chen T, You Y, Jiang H, Wang ZZ (2017) Epithelial–mesenchymal transition (EMT): a biological process in the development, stem cell differentiation, and tumorigenesis. J Cell Physiol 232:3261–3272
Ciocca DR, Calderwood SK (2005) Heat shock proteins in cancer: diagnostic, prognostic, predictive, and treatment implications. Cell Stress Chaperones 10:86–103
Díaz-López A, Moreno-Bueno G, Cano A (2014) Role of microRNA in epithelial to mesenchymal transition and metastasis and clinical perspectives. Cancer Manag Res 6:205–216
Eguchi Y, Shimizu S, Tsujimoto Y (1997) Intracellular ATP levels determine cell death fate by apoptosis or necrosis. Cancer Res 57:1835–1840
Elshamy WM, Duhé RJ (2013) Overview: cellular plasticity, cancer stem cells and metastasis. Cancer Lett 341:2–8
Freed-Pastor WA, Prives C (2012) Mutant p53: one name, many proteins. Genes Dev 26:1268–1286
Gaianigo N, Melisi D, Carbone C (2017) EMT and treatment resistance in pancreatic cancer. Cancers (Basel) 9:E122
Guerra F, Guaragnella N, Arbini AA, Bucci C, Giannattasio S, Moro L (2017) Mitochondrial dysfunction: a novel potential driver of epithelial-to-mesenchymal transition in cancer. Front Oncol 7:295
Huang S, Tang Y, Peng X, Cai X, Wa Q, Ren D, Li Q, Luo J, Li L, Zou X, Huang S (2016) Acidic extracellular pH promotes prostate cancer bone metastasis by enhancing PC-3 stem cell characteristics, cell invasiveness and VEGF-induced vasculogenesis of BM-EPCs. Oncol Rep 36:2025–2032
Kumar S, Stokes J 3rd, Singh UP, Scissum Gunn K, Acharya A, Manne U, Mishra M (2016) Targeting Hsp70: a possible therapy for cancer. Cancer Lett 374:156–166
Lee YJ, Park IS, Lee YJ, Shim JH, Cho MK, Nam HS, Park JW, Oh MH, Lee SH (2014) Resveratrol contributes to chemosensitivity of malignant mesothelioma cells with activation of p53. Food Chem Toxicol 63:153–160
Lee YJ, Bae JH, Kim SA, Kim SH, Woo KM, Nam HS, Cho MK, Lee SH (2017) Cariporide enhances the DNA damage and apoptosis in acid-tolerable malignant mesothelioma H-2452 cells. Mol Cells 40:567–576
Lee SY, Ju MK, Jeon HM, Jeong EK, Lee YJ, Kim CH, Park HG, Han SI, Kang HS (2018) Regulation of tumor progression by programmed necrosis. Oxid Med Cell Longev 2018:3537471
Leu JI, Pimkina J, Frank A, Murphy ME, George DL (2009) A small molecule inhibitor of inducible heat shock protein 70. Mol Cell 36:15–27
Li QQ, Xu JD, Wang WJ, Cao XX, Chen Q, Tang F, Chen ZQ, Liu XP, Xu ZD (2009) Twist1-mediated adriamycin-induced epithelial–mesenchymal transition relates to multidrug resistance and invasive potential in breast cancer cells. Clin Cancer Res 15:2657–2665
Liu T, Daniels CK, Cao S (2012) Comprehensive review on the HSC70 functions, interactions with related molecules and involvement in clinical diseases and therapeutic potential. Pharmacol Ther 136:354–374
Liu J, Bao J, Hao J, Peng Y, Hong F (2014) HSP70 inhibits high glucose-induced Smad3 activation and attenuates epithelial-to-mesenchymal transition of peritoneal mesothelial cells. Mol Med Rep 10:1089–1095
Mattiolo P, Barbero-Farran A, Yuste VJ, Boix J, Ribas J (2014) 2-Phenylethynesulfonamide (PES) uncovers a necrotic process regulated by oxidative stress and p53. Biochem Pharmacol 91:301–311
Mehmood T, Maryam A, Zhang H, Li Y, Khan M, Ma T (2017) Deoxyelephantopin induces apoptosis in HepG2 cells via oxidative stress, NF-κB inhibition and mitochondrial dysfunction. Biofactors 43:63–72
Monma H, Harashima N, Inao T, Okano S, Tajima Y, Harada M (2013) The HSP70 and autophagy inhibitor pifithrin-μ enhances the antitumor effects of TRAIL on human pancreatic cancer. Mol Cancer Ther 12:341–351
Paoli P, Giannoni E, Chiarugi P (2013) Anoikis molecular pathways and its role in cancer progression. Biochim Biophys Acta 1833:3481–3498
Pocaly M, Lagarde V, Etienne G, Ribeil JA, Claverol S, Bonneu M, Moreau-Gaudry F, Guyonnet-Duperat V, Hermine O, Melo JV, Dupouy M, Turcq B, Mahon FX, Pasquet JM (2007) Overexpression of the heat-shock protein 70 is associated to imatinib resistance in chronic myeloid leukemia. Leukemia 21:93–101
Qin X, Ma D, Tan YX, Wang HY, Cai Z (2019) The role of necroptosis in cancer: a double-edged sword? Biochim Biophys Acta Rev Cancer 1871:259–266
Raghunand N, Gillies RJ (2000) pH and drug resistance in tumors. Drug Resist Updat 3:39–47
Ren A, Yan G, You B, Sun J (2008) Down-regulation of mammalian sterile 20-like kinase 1 by heat shock protein 70 mediates cisplatin resistance in prostate cancer cells. Cancer Res 68:2266–2274
Ribas J, Mattiolo P, Boix J (2015) Pharmacological modulation of reactive oxygen species in cancer treatment. Curr Drug Targets 16:31–37
Ricci JE, Muñoz-Pinedo C, Fitzgerald P, Bailly-Maitre B, Perkins GA, Yadava N, Scheffler IE, Ellisman MH, Green DR (2004) Disruption of mitochondrial function during apoptosis is mediated by caspase cleavage of the p75 subunit of complex I of the electron transport chain. Cell 117:773–786
Sekihara K, Harashima N, Tongu M, Tamaki Y, Uchida N, Inomata T, Harada M (2013) Pifithrin-μ, an inhibitor of heat-shock protein 70, can increase the antitumor effects of hyperthermia against human prostate cancer cells. PLoS ONE 8:e78772
Steele AJ, Prentice AG, Hoffbrand AV, Yogashangary BC, Hart SM, Lowdell MW, Samuel ER, North JM, Nacheva EP, Chanalaris A, Kottaridis P, Cwynarski K, Wickremasinghe RG (2009) 2-Phenylacetylenesulfonamide (PAS) induces p53-independent apoptotic killing of B-chronic lymphocytic leukemia (CLL) cells. Blood 114:1217–1225
Strom E, Sathe S, Komarov PG, Chernova OB, Pavlovska I, Shyshynova I, Bosykh DA, Burdelya LG, Macklis RM, Skaliter R, Komarova EA, Gudkov AV (2006) Small-molecule inhibitor of p53 binding to mitochondria protects mice from gamma radiation. Nat Chem Biol 2:474–479
Taylor S, Spugnini EP, Assaraf YG, Azzarito T, Rauch C, Fais S (2015) Microenvironment acidity as a major determinant of tumor chemoresistance: proton pump inhibitors (PPIs) as a novel therapeutic approach. Drug Resist Updat 23:69–78
Wu J, Liu T, Rios Z, Mei Q, Lin X, Cao S (2017) Heat shock proteins and cancer. Trends Pharmacol Sci 38:226–256
Xie T, Peng W, Yan C, Wu J, Gong X, Shi Y (2013) Structural insights into RIP3-mediated necroptotic signaling. Cell Rep 5:70–78
Yang J, Zhu T, Liu X, Zhang L, Yang Y, Zhang J, Guο M (2015) Heat shock protein 70 protects rat peritoneal mesothelial cells from advanced glycation end-products-induced epithelial-to-mesenchymal transition through mitogen-activated protein kinases/extracellular signal-regulated kinases and transforming growth factor-β/Smad pathways. Mol Med Rep 11:4473–4481
Yeh CH, Tseng R, Zhang Z, Cortes J, O’Brien S, Giles F, Hannah A, Estrov Z, Keating M, Kantarjian H, Albitar M (2009) Circulating heat shock protein 70 and progression in patients with chronic myeloid leukemia. Leuk Res 33:212–217
Zhou Y, Ma J, Zhang J, He L, Gong J, Long C (2017) Pifithrin-μ is efficacious against non-small cell lung cancer via inhibition of heat shock protein 70. Oncol Rep 37:313–322
Zhu P, Hu S, Jin Q, Li D, Tian F, Toan S, Li Y, Zhou H, Chen Y (2018) Ripk3 promotes ER stress-induced necroptosis in cardiac IR injury: a mechanism involving calcium overload/XO/ROS/mPTP pathway. Redox Biol 16:157–168
Zou X, Zhang M, Sun Y, Zhao S, Wei Y, Zhang X, Jiang C, Liu H (2015) Inhibitory effects of 3-bromopyruvate in human nasopharyngeal carcinoma cells. Oncol Rep 34:1895–1904
Acknowledgements
This research was supported by the Basic Science Research Program through the National Research Foundation (NRF) of Korea, funded by the Ministry of Education (No. NRF-2018R1D1A1B07046129).
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Dr. Heo attends CnK Pharma but declares that no support, financial or otherwise, has been received from CnK pharma for this research. Other authors state that they have no conflicts of interest.
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Lee, YJ., Park, KS., Heo, SH. et al. Pifithrin-μ induces necroptosis through oxidative mitochondrial damage but accompanies epithelial–mesenchymal transition-like phenomenon in malignant mesothelioma cells under lactic acidosis. Arch. Pharm. Res. 42, 890–901 (2019). https://doi.org/10.1007/s12272-019-01181-6
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DOI: https://doi.org/10.1007/s12272-019-01181-6