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http://dx.doi.org/10.3340/jkns.2019.0187

Dexamethasone Interferes with Autophagy and Affects Cell Survival in Irradiated Malignant Glioma Cells  

Komakech, Alfred (Department of Cancer Control, National Cancer Center Graduate School of Cancer Science and Policy)
Im, Ji-Hye (Department of Cancer Control, National Cancer Center Graduate School of Cancer Science and Policy)
Gwak, Ho-Shin (Department of Cancer Control, National Cancer Center Graduate School of Cancer Science and Policy)
Lee, Kyue-Yim (Department of Cancer Control, National Cancer Center Graduate School of Cancer Science and Policy)
Kim, Jong Heon (Department of Cancer Biomedical Science, National Cancer Center Graduate School of Cancer Science and Policy)
Yoo, Byong Chul (Department of Cancer Biomedical Science, National Cancer Center Graduate School of Cancer Science and Policy)
Cheong, Heesun (Department of Cancer Biomedical Science, National Cancer Center Graduate School of Cancer Science and Policy)
Park, Jong Bae (Department of Cancer Biomedical Science, National Cancer Center Graduate School of Cancer Science and Policy)
Kwon, Ji Woong (Neuro-oncology Clinic, National Cancer Center)
Shin, Sang Hoon (Neuro-oncology Clinic, National Cancer Center)
Yoo, Heon (Neuro-oncology Clinic, National Cancer Center)
Publication Information
Journal of Korean Neurosurgical Society / v.63, no.5, 2020 , pp. 566-578 More about this Journal
Abstract
Objective : Radiation is known to induce autophagy in malignant glioma cells whether it is cytocidal or cytoprotective. Dexamethasone is frequently used to reduce tumor-associated brain edema, especially during radiation therapy. The purpose of the study was to determine whether and how dexamethasone affects autophagy in irradiated malignant glioma cells and to identify possible intervening molecular pathways. Methods : We prepared p53 mutant U373 and LN229 glioma cell lines, which varied by phosphatase and tensin homolog (PTEN) mutational status and were used to make U373 stable transfected cells expressing GFP-LC3 protein. After performing cell survival assay after irradiation, the IC50 radiation dose was determined. Dexamethasone dose (10 μM) was determined from the literature and added to the glioma cells 24 hours before the irradiation. The effect of adding dexamethasone was evaluated by cell survival assay or clonogenic assay and cell cycle analysis. Measurement of autophagy was visualized by western blot of LC3-I/LC3-II and quantified by the GFP-LC3 punctuated pattern under fluorescence microscopy and acridine orange staining for acidic vesicle organelles by flow cytometry. Results : Dexamethasone increased cell survival in both U373 and LN229 cells after irradiation. It interfered with autophagy after irradiation differently depending on the PTEN mutational status : the autophagy decreased in U373 (PTEN-mutated) cells but increased in LN229 (PTEN wild-type) cells. Inhibition of protein kinase B (AKT) phosphorylation after irradiation by LY294002 reversed the dexamethasone-induced decrease of autophagy and cell death in U373 cells but provoked no effect on both autophagy and cell survival in LN229 cells. After ATG5 knockdown, radiation-induced autophagy decreased and the effect of dexamethasone also diminished in both cell lines. The diminished autophagy resulted in a partial reversal of dexamethasone protection from cell death after irradiation in U373 cells; however, no significant change was observed in surviving fraction LN229 cells. Conclusion : Dexamethasone increased cell survival in p53 mutated malignant glioma cells and increased autophagy in PTEN-mutant malignant glioma cell but not in PTEN-wildtype cell. The difference of autophagy response could be mediated though the phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin signaling pathway.
Keywords
Autophagy; Dexamethasone; Glioma; PTEN; Radiation;
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1 Das A, Banik NL, Patel SJ, Ray SK : Dexamethasone protected human glioblastoma U87MG cells from temozolomide induced apoptosis by maintaining Bax:Bcl-2 ratio and preventing proteolytic activities. Mol Cancer 3 : 36,2004   DOI
2 Davis ME : Glioblastoma: overview of disease and treatment. Clin J Oncol Nurs 20(5 Suppl) : S2-S8, 2016   DOI
3 Errafiy R, Aguado C, Ghislat G, Esteve JM, Gil A, Loutfi M, et al. : PTEN increases autophagy and inhibits the ubiquitin-proteasome pathway in glioma cells independently of its lipid phosphatase activity. PLoS One 8 : e83318, 2013   DOI
4 Gewirtz DA : The autophagic response to radiation: relevance for radiation sensitization in cancer therapy. Radiat Res 182 : 363-367, 2014   DOI
5 Gustafsson AB, Gottlieb RA : Recycle or die: the role of autophagy in cardioprotection. J Mol Cell Cardiol 44 : 654-661, 2008   DOI
6 Herzig S, Shaw RJ : AMPK: guardian of metabolism and mitochondrial homeostasis. Nat Rev Mol Cell Biol 19 : 121-135, 2018   DOI
7 Branzei D, Foiani M : Regulation of DNA repair throughout the cell cycle. Nat Rev Mol Cell Biol 9 : 297-308, 2008   DOI
8 Nakamura JL, Karlsson A, Arvold ND, Gottschalk AR, Pieper RO, Stokoe D, et al. : PKB/Akt mediates radiosensitization by the signaling inhibitor LY294002 in human malignant gliomas. J Neurooncol 71 : 215-222, 2005   DOI
9 Oakley RH, Cidlowski JA : The biology of the glucocorticoid receptor: new signaling mechanisms in health and disease. J Allergy Clin Immunol 132 : 1033-1044, 2013   DOI
10 Paglin S, Hollister T, Delohery T, Hackett N, McMahill M, Sphicas E, et al. : A novel response of cancer cells to radiation involves autophagy and formation of acidic vesicles. Cancer Res 61 : 439-444, 2001
11 Pitter KL, Tamagno I, Alikhanyan K, Hosni-Ahmed A, Pattwell SS, Donnola S, et al. : Corticosteroids compromise survival in glioblastoma. Brain 139 : 1458-1471, 2016   DOI
12 Ravikumar B, Sarkar S, Davies JE, Futter M, Garcia-Arencibia M, Green-Thompson ZW, et al. : Regulation of mammalian autophagy in physiology and pathophysiology. Physiol Rev 90 : 1383-1435, 2010   DOI
13 Shields LB, Shelton BJ, Shearer AJ, Chen L, Sun DA, Parsons S, et al. : Dexamethasone administration during definitive radiation and temozolomide renders a poor prognosis in a retrospective analysis of newly diagnosed glioblastoma patients. Radiat Oncol 10 : 222, 2015   DOI
14 Song MS, Salmena L, Pandolfi PP : The functions and regulation of the PTEN tumour suppressor. Nat Rev Mol Cell Biol 13 : 283-296, 2012   DOI
15 Tanida I, Ueno T, Kominami E : LC3 and autophagy. Methods Mol Biol 445 : 77-88, 2008   DOI
16 Tusher VG, Tibshirani R, Chu G : Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci U S A 98 : 5116-5121, 2001   DOI
17 Toth GG, C Kloosterman C, Uges DR, Jonkman MF : Pharmacokinetics of high-dose oral and intravenous dexamethasone. Ther Drug Monit 21 : 532-535, 1999   DOI
18 Inaba H, Pui CH : Glucocorticoid use in acute lymphoblastic leukaemia. Lancet Oncol 11 : 1096-1106, 2010   DOI
19 Toulany M, Lee KJ, Fattah KR, Lin YF, Fehrenbacher B, Schaller M, et al. : Akt promotes post-irradiation survival of human tumor cells through initiation, porgression, and termination of DNA-PKcs-dependent DNA double-strand break repair. Mol Cancer Res 10 : 945-957, 2012   DOI
20 Troncoso R, Paredes F, Parra V, Gatica D, Vasquez-Trincado C, Quiroga C, et al. : Dexamethasone-induced autophagy mediates muscle atrophy through mitochondrial clearance. Cell Cycle 13 : 2281-2295, 2014   DOI
21 Wang Z, Zhou L, Zheng X, Liu W : Effects of dexamethasone on autophagy and apoptosis in acute spinal cord injury. Neuroreport 29 : 1084-1091, 2018   DOI
22 Weller M : Glucocorticoid treatment of primary CNS lymphoma. J Neurooncol 43 : 237-239, 1999   DOI
23 Willers H, Azzoli CG, Santivasi WL, Xia F : Basic mechanisms of therapeutic resistance to radiation and chemotherapy in lung cancer. Cancer J 19 : 200-207, 2013   DOI
24 Wilson EN, Bristol ML, Di X, Maltese WA, Koterba K, Beckman MJ, et al. : A switch between cytoprotective and cytotoxic autophagy in the radiosensitization of breast tumor cells by chloroquine and vitamin D. Horm Cancer 2 : 272-285, 2011   DOI
25 Baskar R, Dai J, Wenlong N, Yeo R, Yeoh KW : Biological response of cancer cells to radiation treatment. Front Mol Biosci 1 : 24, 2014   DOI
26 Aasland D, Reich TR, Tomicic MT, Switzeny OJ, Kaina B, Christmann M : Repair gene O6 -methylguanine-DNA methyltransferase is controlled by SP1 and up-regulated by glucocorticoids, but not by temozolomide and radiation. J Neurochem 144 : 139-151, 2018   DOI
27 Alexanian R, Dimopoulos MA, Delasalle K, Barlogie B : Primary dexamethasone treatment of multiple myeloma. Blood 80 : 887-890, 1992   DOI
28 Andrade MV, Hiragun T, Beaven MA : Dexamethsaone suppresses antigen-induced activation of phosphatidylinositol 3-kinase and downstream responses in mast cells. J Immunol 172 : 7254-7262, 2004   DOI
29 Kim KW, Mutter RW, Cao C, Albert JM, Freeman M, Hallahan DE, et al. : Autophagy for cancer therapy through inhibition of pro-apoptotic proteins and mammalian target of rapamycin signaling. J Biol Chem 281 : 36883-36890, 2006   DOI
30 Jo GH, Bogler O, Chwae YJ, Yoo H, Lee SH, Park JB, et al. : Radiationinduced autophagy contributes to cell death and induces apoptosis partly in malignant glioma cells. Cancer Res Treat 47 : 221-241, 2015   DOI
31 Li HF, Kim JS, Waldman T : Radiation-induced Akt activation modulates radioresistance in human glioblastoma cells. Radiat Oncol 4 : 43, 2009   DOI
32 Luedi MM, Singh SK, Mosley JC, Hassan ISA, Hatami M, Gumin J, et al. : Dexamethasone-mediated oncogenicity in vitro and in an animal model of glioblastoma. J Neurosurg 129 : 1446-1455, 2018   DOI
33 Mattern J, Buchler MW, Herr I : Cell cycle arrest by glucocorticoids may protect normal tissue and solid tumors from cancer therapy. Cancer Biol Ther 6 : 1345-1354, 2007   DOI
34 Milella M, Falcone I, Conciatori F, Cesta Incani U, Del Curatolo A, Inzerilli N, et al. : PTEN: multiple functions in human malignant tumors. Front Oncol 5 : 24, 2015   DOI
35 Molitoris JK, McColl KS, Swerdlow S, Matsuyama M, Lam M, Finkel TH, et al. : Glucocorticoid elevation of dexamethasone-induced gene 2 (Dig2/RTP801/REDD1) protein mediates autophagy in lymphocytes. J Biol Chem 286 : 30181-30189, 2011   DOI
36 Hippert MM, O'Toole PS, Thorburn A : Autophagy in cancer: good, bad, or both? Cancer Res 66 : 9349-9351, 2006   DOI
37 Moretti L, Attia A, Kim KW, Lu B : Crosstalk between Bak/Bax and mTOR signaling regulates radiation-induced autophagy. Autophagy 3 : 142-144, 2007   DOI
38 Cenciarini M, Valentino M, Belia S, Sforna L, Rosa P, Ronchetti S, et al. : Dexamethasone in glioblastoma multiforme therapy: mechanisms and controversies. Front Mol Neurosci 12 : 65, 2019   DOI
39 Yao KC, Komata T, Kondo Y, Kanzawa T, Kondo S, Germano IM : Molecular response of human glioblastoma multiforme cells to ionizing radiation: cell cycle arrest, modulation of the expression of cyclin-dependent kinase inhibitors, and autophagy. J Neurosurg 98 : 378-384, 2003   DOI
40 Yousefi S, Perozzo R, Schmid I, Ziemiecki A, Schaffner T, Scapozza L, et al. : Calpain-mediated cleavage of Atg5 switches autophagy to apoptosis. Nat Cell Biol 8 : 1124-1132, 2006   DOI
41 Chen M, Nowak DG, Trotman LC : Molecular pathways: PI3K pathway phosphatases as biomarkers for cancer prognosis and therapy. Clin Cancer Res 20 : 3057-3063, 2014   DOI
42 Classen F, Kranz P, Riffkin H, Pompsch M, Wolf A, Gopelt K, et al. : Autophagy induced by ionizing radiation promotes cell death over survival in human colorectal cancer cells. Exp Cell Res 374 : 29-37, 2019   DOI
43 Zhang S, Liu Y, Liang Q : Low-dose dexamethasone affects osteoblast viability by inducing autophagy via intracellular ROS. Mol Med Rep 17 : 4307-4316, 2018   DOI