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Functional characteristic of PC12 cells with reduced microsomal glutathione transferase 1

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Microsomal glutathione transferase 1 (MGST1) possesses glutathione transferase and peroxidase activities and is active in biotransformation of xenobiotics and in defense against oxidative stress. To assess MGST1 role in the development and functioning of PC12 cells, we constructed a cell line with reduced MGST1 (PC12_M). Real-time PCR and immunoblot assays showed MGST1 expression lowered to 60 % and immunocytochemical analyses demonstrated an altered concentration and distribution of the enzyme. PC12_M cells revealed a larger tendency to grow in clusters, weaker adhesion, irregular shape of bodies, short neurite outgrowth and higher percentage of necrotic cells (34 %). The total GSTs activity determined with non-specific substrate CDNB (1-chloro-2,4-dinitrobenzene) decreased by 15-20 %, whereas that with DCNB (2,4-dichloro-1-nitrobenzene), a substrate more specific for cytosolic GSTs, was similar to the one in control cells. This suggests that reduction of MGST1 cannot be compensated by other glutathione transferases. In PC12_M cells the total glutathione content was higher by 15-20 %, whereas the GSSG/GSH ratio was lower than in control cells. Moreover, the laminin-dependent migration rate was much faster in control cells than in PC12_M, suggesting some alterations in the metastatic potential of the line with suppressed MGST1. The amount of MAP kinases (p38, JNK, ERK1/2) was elevated in PC12_M cells but their phosphorylation level declined. Microarray analysis showed changed expression of several genes, which may be linked with differentiation and necrosis of PC12_M cells. Our data suggest that MGST1 could be an important regulator of PC12 cells development and might have significant effects on cell growth and proliferation, probably through altered expression of genes with different biological function.
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  • Department of Molecular Neurochemistry, Medical University, Łódź, Poland
  • Department of Molecular Neurochemistry, Medical University, Łódź, Poland
  • Department of Molecular Neurochemistry, Medical University, Łódź, Poland
  • Department of Molecular Neurochemistry, Medical University, Łódź, Poland
  • Department of Molecular Neurochemistry, Medical University, Łódź, Poland
  • Department of Molecular and Medical Biophysics, Medical University, Łódź, Poland
  • Department of Molecular and Medical Biophysics, Medical University, Łódź, Poland
  • Department of Molecular and Medical Biophysics, Medical University, Łódź, Poland
  • Department of Molecular Neurochemistry, Medical University, Łódź, Poland
  • Bagheri-Yarmand R, Mazumdar A, Sahin AA, Kumar R (2006) LIM kinase 1 increases tumor metastasis of human breast cancer cells via regulation of the urokinase-type plasminogen activator system. Int J Cancer 118: 2703-2710.
  • Bertrand MJM, Kenchappa RS, Andrieu D, Leclerq-Smekens M, Nguyen HNT, Carter BD, Muscatelli F, Barker PA, De Backer O (2008) NRAGE, a p75NTR adaptor protein, is required for developmental apoptosis in vivo. Cell Death Differ 15: 1921-1929.
  • Dhariwala FA and Rajadhyaksha MS (2008) An unusual member of the Cdk family: Cdk5. Cell Mol Neurobiol 28: 351-369.
  • Davila M, Frost AR, Grizzle WE, Chakrabarti R (2003) LIM Kinase 1 is essential for the invasive growth of prostate epithelial cells. J Biol Chem 278: 36868-36875.
  • Di Paolo G, Pellier V, Catsicas M, Antonsson B, Catsicas S, Grenningloh G (1996) The phosphoprotein stathmin is essential for nerve growth factor-stimulated differentiation. J Cell Biol 133: 1383-1390.
  • Estonius M, Forsberg L, Danielsson O, Weinander R, Kelner MJ, Morgenstern R (1999) Distribution of microsomal glutathione transferase 1 in mammalian tissue. A predominant alternate first exon in human tissues. Eur J Biochem 260: 409-413.
  • Filipek A, Michowski W, Kuznicki J (2007) Involvement of S100A6 (calcyclin) and its binding partners in intracellular signaling pathways. Adv Enzyme Regul 48: 225-239.
  • Golstein P, Kroemer G (2006) Cell death by necrosis: towards a molecular definition. Trends Biochem Sci 32: 37-43.
  • Habig WH, Jakoby WB (1981) Assays for differentiation of glutathione-S-transferases. Methods Enzymol 77: 398-405.
  • Hu F, Strittmatter SM (2008) The N-terminal domain of nogo-a inhibits cell adhesion and axonal outgrowth by an integrin-specific mechanism. J Neurosci 28: 1262-1269.
  • Imaizumi N, Miyagi S, Aniya Y (2006) Reactive nitrogen species derived activation of rat liver microsomal glutatione S-transferase. Life Sci 78: 2998-3006.
  • Jakobbson Per-Johan, Morgenstern R, Mancini J, Ford-Hutchinson A, Persson B (2000) Membrane-associated proteins in eicosanoid and glutathione metabolism (MAPEG). a widespread protein superfamily. Am J Respir Crit Care Med 161: 20-24.
  • Jezek P, Hlavatá L (2005) Mitochondria in homeostasis of reactive oxygen species in cell, tissues, and organism. Int J Biochem Cell Biol 37: 2478-503.
  • Johansson K, Åhlen K, Rinaldi R, Sahlander K, Siritantikorn A and Morgenstern R (2007) Microsomal glutathione transferase 1 in anticancer drug resistance. Carcinogenesis 28: 465-470.
  • Kim EK, Choi E-J (2010) Pathological roles of MAPK signaling pathways in human diseases. Biochim Biophys Acta 1802: 396-405.
  • Lambeng N. Michel PP, Agid Y, Ruberg M (2001) The relationship between differentiation and survival in PC12 cells treated with cyclic adenosine monophosphate in the presence of epidermal growth factor or nerve growth factor. Neurosci Lett 297: 133-136.
  • Lee SH, DeJong J (1999) Microsomal GST-I: genomic organization, expression, and alternative splicing of the human gene. Biochim Biophys Acta 1446: 389-396.
  • Lee KK, Shimoji M, Hossain QS, Sunakawa H, Aniya Y (2008) Novel function of glutathione transferase in rat liver mitochondrial membrane: Role for cytochrome c release from mitochondria. Toxicol Appl Pharmacol 232: 109-118.
  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method. Methods 25: 402-408.
  • Mistry SJ, Atweh GF (2002) Role of stathmin in the regulation of the mitotic spindle: potential applications in cancer therapy. Mt Sinai J Med 69: 299-305.
  • Oiwa Y, Yoshimura R, Nakai k, Itakura T (2002) Dopaminergic neuroprotection and regeneration by neurturin assessed by using behavioral, biochemical and histochemical measurements in a model of progressive Parkinson's disease. Brain Res 947: 271-283.
  • Rankin SL, Rahimtula M, Mearow KM (2006) A method to assess multiple aspects of the motile behaviour of adherent PC12 cells on applied biological substrates. J Neurosci Methods 156: 55-63.
  • Raveh S, Gavert N, Ben-Ze'ev A (2009) L1 cell adhesion molecule (L1cam) in invasive tumors. Cancer Lett 282: 137-145.
  • Raza H, John A (2006) 4-hydroxynonenal induces mitochondrial oxidative stress, apoptosis and expression of glutathione S-transferase A4-4 and cytochrome P450 2E1 in PC12 cells. Toxicol Appl Pharmacol 216: 309-18.
  • Scott RW, Olson MF (2007) LIM kinases: function, regulation and association with human disease. J Mol Med 85: 555-568.
  • Shi H, Lu D, Shu Y, Shi W, Lu S, Wang K (2008) Expression of multidrug-resistance-related proteins p-glycoprotein, glutathione-s-transferases, topoisomerase-II and lung resistance protein in primary gastric cardiac adenocarcinoma. Cancer Invest 26: 344-351.
  • Siritantikorn A, Johansson K, Åhlen K, Rinaldi R, Suthiphongchai T, Wilairat P, Morgenstern R (2007) Protection of cells from oxidative stress by Microsomal glutathione transferase 1. Biochem Biophys Res Commun 355: 592-596.
  • Tehranian R, Rose ME, Vagni V, Pickrell AM, Griffith RP, Liu H, Clark RSB, Edward Dixon C, Kochanek PM, Graham SH (2008) Disruption of bax protein prevents neuronal cell death but produces cognitive impairment in mice following traumatic brain injury. J Neurotrauma 25: 755-767.
  • Tomaselli KJ, Damsky CH, Reichardt LF (1987) Interactions of a neuronal cell line (PC12) with laminin, collagen iv, and fibronectin: identification of integrin-related glycoproteins involved in attachment and process outgrowth. J Cell Biol 105: 2347-2358.
  • Tripathi BK, Zelenka PS (2010) Cdk5. A regulator of epithelial cell adhesion and migriation. Cell Adh Migr 4: 333-336.
  • Uttara B, Singh AV, Zamboni P, Mahajan RT (2009) Oxidative stress and neurodegenerative diseases: a review of upstream and downstream antioxidant therapeutic options. Curr Neuropharmacol 7: 65-74.
  • Von Bernhardi R, Tichauer JE, Eugenin J (2010) Aging-dependent changes of microglial cells and their relevance for neurodegenerative disorders. J Neurochem 112: 1099-1114.
  • Wagner EF, Nebreda AR (2009) Signal integration by JNK and p38 MAPK pathways in cancer development. Nat Rev Cancer 9: 537-549.
  • Wu G, Fang Y-Z, Yang S, Lupton JR, Turner ND (2004) Glutathione metabolism and its implications for health. J Nutr 134: 489-492.
  • Wykosky J, Debinski W (2008) The EphA2 receptor and ephrinA1 ligand in solid tumors: function and therapeutic targeting. Mol Cancer Res 6: 1795-806.
  • Yanbin J, Neverova I, Van Eyk JE, Bennett BM (2006) Nitration of tyrosine 92 mediates the activation of rat microsomal glutathione S-transferase by peroxynitrite. J Biol Chem 281: 1986-1991.
  • Yang Y, Sharma R, Sharma A, Awasthi S, Awasthi YC (2003) Lipid peroxidation and cell cycle signaling: 4-hydroxynonenal, a key molecule in stress mediated signaling. Acta Biochim Pol 50: 319-336.
  • Yang YS, Strittmatter SM (2007) The reticulons: a family of proteins with diverse functions. Genome Biol 8: 234.
  • Zylinska L, Kozaczuk A, Szemraj J, Kargas Ch, Kowalska I (2007) Functional importance of PMCA isoforms in growth and development of PC12 Cells. Ann NY Acad Sci 1099: 254-269.
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