1. bookVolume 50 (2016): Issue 3 (July 2016)
Journal Details
First Published
30 Mar 2016
Publication timeframe
4 times per year
Open Access

Inhibition of IRE1 signaling affects the expression of genes encoded glucocorticoid receptor and some related factors and their hypoxic regulation in U87 glioma cells

Published Online: 20 Aug 2016
Volume & Issue: Volume 50 (2016) - Issue 3 (July 2016)
Page range: 127 - 136
Journal Details
First Published
30 Mar 2016
Publication timeframe
4 times per year

Objective. The aim of the present investigation was to examine the effect of inhibition of endoplasmic reticulum stress signaling, mediated by IRE1 (inositol requiring enzyme 1), which is a central mediator of the unfolded protein response on the expression of genes encoding glucocorticoid receptor (NR3C1) and some related proteins (SGK1, SGK3, NCOA1, NCOA2, ARHGAP35, NNT) and their hypoxic regulation in U87 glioma cells for evaluation of their possible significance in the control of the glioma growth.

Methods. The expression of NR3C1,SGK1,SGK3, NCOA1, NCOA2, ARHGAP35, and NNT genes in U87 glioma cells, transfected by empty vector pcDNA3.1 (control) and cells without IRE1 signaling enzyme function (transfected by dnIRE1) upon hypoxia, was studied by quantitative polymerase chain reaction.

Results. Inhibition of IRE1 signaling enzyme function up-regulates the expression of NR3C1, SGK1, NCOA1, NCOA2, ARHGAP35, and NNT genes in U87 glioma cells in comparison with the control glioma cells, with more significant changes for NR3C1, SGK1, and NNT genes. At the same time, the expression of SGK3 gene is strongly down-regulated in glioma cells upon inhibition of IRE1. We have also shown that hypoxia increases the expression of NR3C1, SGK1, NCOA2, ARHGAP35, and NNT genes but decreases SGK3 and NCOA1 genes expression in control glioma cells. Moreover, the inhibition of both enzymatic activities (kinase and endoribonuclease) of IRE1 in U87 glioma cells enhances the eff ect of hypoxia on the expression of SGK1, SGK3, and NNT genes, but decreases the sensitivity of NR3C1 gene to hypoxic condition. Furthermore, the expression of NCOA1 gene is resistant to hypoxia in control glioma cells, but NCOA2 and ARHGAP35 genes are resistant to this condition in glioma cells without functional activity of IRE1 signaling enzyme.

Conclusions. Results of this investigation demonstrate that inhibition of IRE1 signaling enzyme function affects the expression of NR3C1, SGK1, SGK3, NCOA1, NCOA2, ARHGAP35, and NNT genes in U87 glioma cells in gene specific manner and that all these genes are regulated by hypoxia preferentially through IRE1 signaling pathway of the endoplasmic reticulum stress.


Acosta-Alvear D, Zhou Y, Blais A, Tsikitis M, Lents NH, Arias C, Lennon CJ, Kluger Y, Dynlacht DD. XBP1 controls diverse cell type- and condition-specific transcriptional regulatory networks. Molecular Cell 27, 53-66, 2007.10.1016/j.molcel.2007.06.01117612490Search in Google Scholar

Aragon T, van Anken E, Pincus D, Serafimova IM, Korennykh AV, Rubio CA, Walter P. Messenger RNA targeting to endoplasmic reticulum stress signalling sites. Nature 457, 736-740, 2009.10.1038/nature07641276853819079237Search in Google Scholar

Auf G, Jabouille A, Guerit S, Pineau R, Delugin M, Bouchecareilh M, Favereaux A, Maitre M, Gaiser T, von Deimling A, Czabanka M, Vajkoczy P, Chevet E, Bikfalvi A, Moenner M. A shift from an angiogenic to invasive phenotype induced in malignant glioma by inhibition of the unfolded protein response sensor IRE1. Proc Natl Acad Sci USA 107, 15553-15558, 2010.10.1073/pnas.0914072107293260020702765Search in Google Scholar

Auf G, Jabouille A, Delugin M, Guerit S, Pineau R, North S, Platonova N, Maitre M, Favereaux A, Vajkoczy P, Seno M, Bikfalvi A, Minchenko D, Minchenko O, Moenner M. High epiregulin expression in human U87 glioma cells relies on IRE1alpha and promotes autocrine growth through EGF receptor. BMC Cancer 13, 597, 2013.10.1186/1471-2407-13-597387867024330607Search in Google Scholar

Bochkov VN, Philippova M, Oskolkova O, Kadl A, Furnkranz A, Karabeg E, Breuss J, Minchenko OH, Mechtcheriakova D, Hohensinner P, Rychli K, Wojta J, Resink T, Binder BR, Leitinger N. Oxidized phospholipids stimulate angiogenesis via induction of VEGF, IL-8, COX-2 and ADAMTS-1 metalloprotease, implicating a novel role for lipid oxidation in progression and destabilization of atherosclerotic lesions. Circ Res 99, 900-908, 2006.10.1161/01.RES.0000245485.04489.ee16973904Search in Google Scholar

Bravo R, Parra V, Gatica D, Rodriguez AE, Torrealba N, Paredes F, Wang ZV, Zorzano A, Hill JA, Jaimovich E, Quest AF, Lavandero S. Endoplasmic reticulum and the unfolded protein response: dynamics and metabolic integration. Int Rev Cell Mol Biol 301, 215-290, 2013.10.1016/B978-0-12-407704-1.00005-1366655723317820Search in Google Scholar

Cao SS, Kaufman RJ. Targeting endoplasmic reticulum stress in metabolic disease. Expert Opin Th er Targets 17, 437-448, 2013.10.1517/14728222.2013.75647123324104Search in Google Scholar

Chevet E, Hetz C, Samali A. Endoplasmic reticulum stress-activated cell reprogramming in oncogenesis. Cancer Discov 5, 586-597, 2015.10.1158/2159-8290.CD-14-149025977222Search in Google Scholar

Chi MN, Guo ST, Wilmott JS, Guo XY, Yan XG, Wang CY, Liu XY, Jin L, Tseng HY, Liu T, Croft A, Hondermarck H, Scolyer RA, Jiang CC, Zhang XD. INPP4B is upregulated and functions as an oncogenic driver through SGK3 in a subset of melanomas. Oncotarget 6, 39891-39907, 2015.10.18632/oncotarget.5359474186826573229Search in Google Scholar

Clarke HJ, Chambers JE, Liniker E, Marciniak SJ. Endoplasmic reticulum stress in malignancy. Cancer Cell 25, 563-573, 2014.10.1016/j.ccr.2014.03.01524823636Search in Google Scholar

Dasgupta S, Putluri N, Long W, Zhang B, Wang J, Kaushik AK, Arnold JM, Bhowmik SK, Stashi E, Brennan CA, Rajapakshe K, Coarfa C, Mitsiades N, Ittmann MM, Chinnaiyan AM, Sreekumar A, O’Malley BW. Coactivator SRC-2-dependent metabolic reprogramming mediates prostate cancer survival and metastasis. J Clin Invest 125, 1174-1188, 2015. 10.1172/JCI76029436226025664849Search in Google Scholar

de Guia RM, Herzig S. How do glucocorticoids regulate lipid metabolism? Adv Exp Med Biol 872, 127-144, 2015.10.1007/978-1-4939-2895-8_626215993Search in Google Scholar

Dejeans N, Pluquet O, Lhomond S, Grise F, Bouchecareilh M, Juin A, Meynard-Cadars M, Bidaud-Meynard A, Gentil C, Moreau V, Saltel F, Chevet E. Autocrine control of glioma cells adhesion and migration through IRE1a-mediated cleavage of SPARC mRNA. J Cell Sci 125, 4278-4287, 2012.Search in Google Scholar

Dejeans N, Barroso K, Fernandez-Zapico ME, Samali A, Chevet. Novel roles of the unfolded protein response in the control of tumor development and aggressiveness. Semin Cancer Biol 33, 67-73, 2015.10.1016/j.semcancer.2015.04.00725953433Search in Google Scholar

Drogat B, Auguste P, Nguyen DT, Bouchecareilh M, Pineau R, Nalbantoglu J, Kaufman RJ, Chevet E, Bikfalvi A, Moenner M. IRE1 signaling is essential for ischemia-induced vascular endothelial growth factor-A expression and contributes to angiogenesis and tumor growth in vivo. Cancer Res 67, 6700-6707, 2007.10.1158/0008-5472.CAN-06-323517638880Search in Google Scholar

Fenne IS, Helland T, Flageng MH, Dankel SN, Mellgren G, Sagen JV. Downregulation of steroid receptor coactivator-2 modulates estrogen-responsive genes and stimulates proliferation of mcf-7 breast cancer cells. PLoS ONE 8, E70096, 2013.10.1371/journal.pone.0070096372835723936147Search in Google Scholar

Han J, Back SH, Hur J, Lin YH, Gildersleeve R, Shan J, Yuan CL, Krokowski D, Wang S, Hatzoglou M, Kilberg MS, Sartor MA, Kaufman RJ. ER-stress-induced transcriptional regulation increases protein synthesis leading to cell death. Nat Cell Biol 15, 481-490, 2013.10.1038/ncb2738369227023624402Search in Google Scholar

Han J, Kaufman RJ. Measurement of the unfolded protein response to investigate its role in adipogenesis and obesity. Methods Enzymol 538,135-150, 2014.10.1016/B978-0-12-800280-3.00008-624529437Search in Google Scholar

Hausmann S, Brandt E, Kochel C, Einsele H, Bargou RC, Seggewiss-Bernhardt R, Stuhmer T. Loss of serum and glucocorticoid-regulated kinase 3 (SGK3) does not affect proliferation and survival of multiple myeloma cell lines. PLoS One 10, e0122689, 2015.10.1371/journal.pone.0122689438354525837824Search in Google Scholar

Heiker JT, Kern M, Kosacka J, Flehmig G, Stumvoll M, Shang E, Lohmann T, Dressler M, Kovacs P, Bluher M, Kloting N. Nicotinamide nucleotide transhydrogenase mRNA expression is related to human obesity. Obesity (Silver Spring) 21, 529-534, 2013.10.1002/oby.2009523592659Search in Google Scholar

Huang Y, Zhou J, Huang Y, He J, Wang Y, Yang C, Liu D, Zhang L, He F. SARI, a novel target gene of glucocorticoid receptor, plays an important role in dexamethasone-mediated killing of B lymphoma cells. Cancer Lett 373, 57-66, 2016.10.1016/j.canlet.2016.01.03426808579Search in Google Scholar

Khan SH, Ling J, Kumar R. TBP binding-induced folding of the glucocorticoid receptor AF1 domain facilitates its interaction with steroid receptor coactivator-1. PLoS ONE 6, E21939, 2011.10.1371/journal.pone.0021939313138521760925Search in Google Scholar

Kim IK, Kim BS, Koh CH, Seok JW, Park JS, Shin KS, Bae EA, Lee GE, Jeon H, Cho J, Jung Y, Han D, Kwon BS, Lee HY, Chung Y, Kang CY. Glucocorticoid-induced tumor necrosis factor receptor-related protein co-stimulation facilitates tumor regression by inducing IL-9-producing helper T cells. Nat Med 21, 1010-1017, 2015.10.1038/nm.392226280119Search in Google Scholar

Langlais P, Yi Z, Finlayson J, Luo M, Mapes R, De Filippis E, Meyer C, Plummer E, Tongchinsub P, Mattern M, Mandarino LJ. Global IRS-1 phosphorylation analysis in insulin resistance. Diabetologia 54, 2878-2889, 2011.10.1007/s00125-011-2271-9388216521850561Search in Google Scholar

Lee J, Ozcan U. Unfolded protein response signaling and metabolic diseases. J Biol Chem 289, 1203-1211, 2014.10.1074/jbc.R113.534743389430624324257Search in Google Scholar

Lenihan CR, Taylor CT. Th e impact of hypoxia on cell death pathways. Biochem Soc Trans 41, 657-663, 2013.10.1042/BST2012034523514172Search in Google Scholar

Li MD, Ruan HB, Singh JP, Zhao L, Zhao T, Azarhoush S, Wu J, Evans RM, Yang X. O-GlcNAc transferase is involved in glucocorticoid receptor-mediated transrepression. J Biol Chem 287, 12904-12912, 2012.10.1074/jbc.M111.303792333997022371499Search in Google Scholar

Liu ST, Lu GY, Hsu YJ, Chang LC, Ho CL, Huang SM. Dual roles for lysine 490 of promyelocytic leukemia protein in the transactivation of glucocorticoid receptor-interacting protein 1. Biochim Biophys Acta 1833, 1799-1810, 2013.10.1016/j.bbamcr.2013.03.01523542129Search in Google Scholar

Liu H, Li C, Shen C, Yin F, Wang K, Liu Y, Zheng B, Zhang W, Hou X, Chen X, Wu J, Wang X, Zhong C, Zhang J, Shi H, Ai J, Zhao S. MiR-212-3p inhibits glioblastoma cell proliferation by targeting SGK3. J Neurooncol 122, 431-439, 2015.10.1007/s11060-015-1736-y25720694Search in Google Scholar

Manie SN, Lebeau J, Chevet E. Cellular mechanisms of endoplasmic reticulum stress signaling in health and disease. 3. Orchestrating the unfolded protein response in oncogenesis: an update. Am J Physiol Cell Physiol 307, C901-C907, 2014.10.1152/ajpcell.00292.201425186011Search in Google Scholar

Maurel M, Chevet E, Tavernier J, Gerlo S. Getting RIDD of RNA: IRE1 in cell fate regulation. Trends Biochem Sci 39, 245-254, 2014.10.1016/j.tibs.2014.02.00824657016Search in Google Scholar

Maurel M, McGrath EP, Mnich K, Healy S, Chevet E, Samali A. Controlling the unfolded protein response-mediated life and death decisions in cancer. Semin Cancer Biol 33, 57-66, 2015.10.1016/j.semcancer.2015.03.00325814342Search in Google Scholar

Meimaridou E, Kowalczyk J, Guasti L, Hughes CR, Wagner F, Frommolt P, Nurnberg P, Mann NP, Banerjee R, Saka HN, Chapple JP, King PJ, Clark AJ, Metherell LA. Mutations in NNT encoding nicotinamide nucleotide transhydrogenase cause familial glucocorticoid deficiency. Nat Genet 44, 740-742, 2012.10.1038/ng.2299338689622634753Search in Google Scholar

Minchenko AG, Germanyuk YL. Effect of hydrocortisone on the expression of mitochondrial genes in the liver of normal and alloxan diabetic rats. Endocrinol Exper 18, 3-18, 1984. Search in Google Scholar

Minchenko AG. Eff ect of hydrocortisone on biosynthesis of mitochondrial and cytoplasmic RNA in liver of adrenalectomized rats. Endocrinol Exper 22, 75-86, 1988.Search in Google Scholar

Minchenko AG, Tronjko ND. Subcellular distribution of 3H-hydrocortisone and its metabolites in the liver and kidneys of normal and alloxan diabetic rats. Endocrinol Exper 22, 19-28, 1988.Search in Google Scholar

Minchenko OH, Kharkova AP, Bakalets TV, Kryvdiuk IV. Endoplasmic reticulum stress, its sensor and signaling systems and the role in regulation of gene expressions at malignant tumor growth and hypoxia. Ukr Biokhim Zh 85, 5-16, 2013a.10.15407/ubj85.05.00524479318Search in Google Scholar

Minchenko DO, Kharkova AP, Hubenia OV, Minchenko OH. Insulin receptor, IRS1, IRS2, INSIG1, INSIG2, RRA D, and BAIAP2 gene expressions in glioma U87 cells with ERN1 loss of function: effect of hypoxia and glutamine or glucose deprivation. Endocr Regul 47, 15-26, 2013b.10.4149/endo_2013_01_1523363253Search in Google Scholar

Minchenko DO, Danilovskyi SV, Kryvdiuk IV, Bakalets TV, Lypova NM, Karbovskyi LL, Minchenko OH. Inhibition of ERN1 modifies the hypoxic regulation of the expression of TP53-related genes in U87 glioma cells. Endoplasm Reticul Stress Dis 1, 18-26, 2014.10.2478/ersc-2014-0001Search in Google Scholar

Minchenko DO, Kharkova AP, Tsymbal DO, Karbovskyi LL, Minchenko OH. Expression of insulin-like growth factor binding protein genes and its hypoxic regulation in U87 glioma cells depends on ERN1 mediated signaling pathway of endoplasmic reticulum stress. Endocr Regul 49, 73-83, 2015a.10.4149/endo_2015_02_7325960008Search in Google Scholar

Minchenko OH, Tsymbal DO, Moenner M, Minchenko DO, Kovalevska OV, Lypova NM. Inhibition of the endoribonuclease of ERN1 signaling enzyme affects the expression of proliferation-related genes in U87 glioma cells. Endoplasm Reticul Stress Dis 2, 18-29, 2015b.10.1515/ersc-2015-0002Search in Google Scholar

Notsuda H, Sakurada A, Endo C, Okada Y, Horii A, Shima H, Kondo T. p190A RhoGAP is involved in EGFR pathways and promotes proliferation, invasion and migration in lung adenocarcinoma cells. Int J Oncol 43, 1569-1577, 2013.10.3892/ijo.2013.209624043274Search in Google Scholar

Organ SL, Hai J, Radulovich N, Marshall CB, Leung L, Sasazuki T, Shirasawa S, Zhu CQ, Navab R, Ikura M, Tsao MS. p120RasGAP is a mediator of rho pathway activation and tumorigenicity in the DLD1 colorectal cancer cell line. PLoS One 9, e86103, 2014.10.1371/journal.pone.0086103389762224465899Search in Google Scholar

Pluquet O, Dejeans N, Bouchecareilh M, Lhomond S, Pineau R, Higa A, Delugin M, Combe C, Loriot S, Cubel G, Dugot-Senant N, Vital A, Loiseau H, Gosline SJ, Taouji S, Hallett M, Sarkaria JN, Anderson K, Wu W, Rodriguez FJ, Rosenbaum J, Saltel F, Fernandez-Zapico ME, Chevet E. Posttranscriptional regulation of PER1 underlies the oncogenic function of IREα. Cancer Res 73, 4732-4743, 2013.10.1158/0008-5472.CAN-12-3989391571623752693Search in Google Scholar

Pluquet O, Dejeans N, Chevet E. Watching the clock: endoplasmic reticulum-mediated control of circadian rhythms in cancer. Ann Med 46, 233-243, 2014.10.3109/07853890.2013.87466424491143Search in Google Scholar

Psarra AM, Sekeris CE. Glucocorticoids induce mitochondrial gene transcription in HepG2 cells: role of the mitochondrial glucocorticoid receptor. Biochim Biophys Acta 1813, 1814-1821, 2011.10.1016/j.bbamcr.2011.05.01421664385Search in Google Scholar

Pufall MA. Glucocorticoids and cancer. Adv Exp Med Biol 872, 315-333, 2015.10.1007/978-1-4939-2895-8_14554609926216001Search in Google Scholar

Qin J, Chen JX, Zhu Z, Teng JA. Genistein inhibits human colorectal cancer growth and suppresses miR-95, Akt and SGK1. Cell Physiol Biochem 35, 2069-2077, 2015.10.1159/00037401325871428Search in Google Scholar

Sheeran FL, Rydstrom J, Shakhparonov MI, Pestov NB, Pepe S. Diminished NADPH transhydrogenase activity and mitochondrial redox regulation in human failing myocardium. Biochim Biophys Acta 1797, 1138-1148, 2010.10.1016/j.bbabio.2010.04.00220388492Search in Google Scholar

Srivastava M, Leighton X, Starr J, Eidelman O, Pollard HB. Diverse effects of ANXA7 and p53 on LNCaP prostate cancer cells are associated with regulation of SGK1 transcription and phosphorylation of the SGK1 target FOXO3A. Biomed Res Int 2014, 193635, 2014.10.1155/2014/193635401690724864229Search in Google Scholar

Thomas AL, Coarfa C, Qian J, Wilkerson JJ, Rajapakshe K, Krett NL, Gunaratne PH, Rosen ST. Identification of potential glucocorticoid receptor therapeutic targets in multiple myeloma. Nucl Recept Signal 13, e006, 2015.10.1621/nrs.13006469362926715915Search in Google Scholar

Vonlanthen J, Okoniewski MJ, Menigatti M, Cattaneo E, Pellegrini-Ochsner D, Haider R, Jiricny J, Staiano T, Buffoli F, Marra G. A comprehensive look at transcription factor gene expression changes in colorectal adenomas. BMC Cancer 14, 46, 2014.10.1186/1471-2407-14-46407800524472434Search in Google Scholar

Xiaobo Y, Qiang L, Xiong Q, Zheng R, Jianhua Z, Zhifeng L, Yijiang S, Zheng J. Serum and glucocorticoid kinase 1 promoted the growth and migration of non-small cell lung cancer cells. Gene 576, 339-346, 2016.10.1016/j.gene.2015.10.07226548813Search in Google Scholar

Zhao J, Xu H, He M, Wu Y. Glucocorticoid receptor DNA binding factor 1 expression and osteosarcoma prognosis. Tumour Biol 35, 12449-12458, 2014. 10.1007/s13277-014-2563-z25185653Search in Google Scholar

Recommended articles from Trend MD

Plan your remote conference with Sciendo