Multiple roles of microsomal glutathione transferase 1 in cellular protection: a mechanistic study.

The aim of this study was to investigate the involvement of membrane-bound microsomal glutathione transferase 1 (MGST1) in cellular resistance against oxidative stress as well as its mechanism of protection. MGST1 is ubiquitously expressed and predominantly located in the endoplasmic reticulum and outer mitochondrial membrane. Utilizing MCF7 cells overexpressing MGST1 we show significant protection against agents that are known to induce lipid peroxidation (e.g., cumene hydroperoxide and tert-butylhydroperoxide) and an end-product of lipid peroxidation (e.g., 4-hydroxy-2-nonenal). Furthermore, our results demonstrate that MGST1 protection can be enhanced by vitamin E when toxicity depends on oxidative stress, but not when direct alkylation is the dominant mechanism. Mitochondria in MGST1-overexpressing cells were shown to be protected from oxidative insult as measured by calcium loading capacity and respiration. MGST1 induces cellular resistance against cisplatin. Here we used vitamin E to elucidate whether oxidative stress caused by cisplatin is significant for cell toxicity. The results indicate that oxidative stress and induction of lipid peroxidation are not the most prominent toxic mechanism of cisplatin in our cell system. We thus conclude that MGST1 protects cells (and mitochondria) by both conjugation and glutathione peroxidase functions. A new protective mechanism against cisplatin is also indicated.

[1]  R. Burk,et al.  Influence of vitamin E and selenium on glutathione-dependent protection against microsomal lipid peroxidation. , 1984, Biochemical pharmacology.

[2]  Nikola Kasabov,et al.  Gene Expression Proé ling of Metastatic and Nonmetastatic Colorectal Cancer Cell Lines , 2002 .

[3]  J. Hayes,et al.  Potential contribution of the glutathione S-transferase supergene family to resistance to oxidative stress. , 1995, Free radical research.

[4]  R. Moorehead,et al.  Use of a transgenic mouse model to identify markers of human lung tumors , 2005, International journal of cancer.

[5]  H. H. Draper,et al.  Malondialdehyde determination as index of lipid peroxidation. , 1990, Methods in enzymology.

[6]  H. Barnes,et al.  Heterologous expression of rat liver microsomal glutathione transferase in simian COS cells and Escherichia coli. , 1995, The Biochemical journal.

[7]  K Arvidson,et al.  Cytotoxic and genotoxic effects of areca nut-related compounds in cultured human buccal epithelial cells. , 1989, Cancer research.

[8]  Ralf Morgenstern,et al.  Protection of cells from oxidative stress by microsomal glutathione transferase 1. , 2007, Biochemical and biophysical research communications.

[9]  R. Morgenstern,et al.  Microsomal glutathione transferase 1 in anticancer drug resistance. , 2006, Carcinogenesis.

[10]  M. Saito,et al.  Early embryonic lethality caused by targeted disruption of the mouse PHGPx gene. , 2003, Biochemical and biophysical research communications.

[11]  W B Jakoby,et al.  Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. , 1974, The Journal of biological chemistry.

[12]  R. Morgenstern,et al.  The distribution of microsomal glutathione transferase among different organelles, different organs, and different organisms. , 1984, Biochemical pharmacology.

[13]  C. C. Reddy,et al.  Vitamin E dependent reduced glutathione inhibition of rat liver microsomal lipid peroxidation. , 1982, Life sciences.

[14]  R. Morgenstern,et al.  Evidence that rat liver microsomal glutathione transferase is responsible for glutathione-dependent protection against lipid peroxidation. , 1993, Biochemical pharmacology.

[15]  K. Palczewski,et al.  Microsomal glutathione S-transferase 1 in the retinal pigment epithelium: protection against oxidative stress and a potential role in aging. , 2005, Biochemistry.

[16]  A. Dafre,et al.  Protein S-thiolation and redox regulation of membrane-bound glutathione transferase. , 1998, Chemico-biological interactions.

[17]  E. Ajandouz,et al.  Alpha-tocopheryl acetate is absorbed and hydrolyzed by Caco-2 cells comparative studies with alpha-tocopherol. , 2008, Chemistry and physics of lipids.

[18]  Aniya Yoko,et al.  Oxidative stress-induced activation of microsomal glutathione S-transferase in isolated rat liver , 1993 .

[19]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[20]  P. van Bladeren,et al.  Microsomal glutathione transferase: lipid-derived substrates and lipid dependence. , 1995, Archives of biochemistry and biophysics.

[21]  M. Matzuk,et al.  Neurodegeneration, myocardial injury, and perinatal death in mitochondrial superoxide dismutase-deficient mice. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[22]  M Crompton,et al.  The mitochondrial permeability transition pore and its role in cell death. , 1999, The Biochemical journal.

[23]  A. Nègre-Salvayre,et al.  Advanced lipid peroxidation end products in oxidative damage to proteins. Potential role in diseases and therapeutic prospects for the inhibitors , 2008, British journal of pharmacology.

[24]  Y. Tsujimoto,et al.  Mitochondrial membrane permeability transition and cell death. , 2006, Biochimica et biophysica acta.

[25]  R. Morgenstern,et al.  Activation of microsomal glutathione S-transferase activity by sulfhydryl reagents. , 1979, Biochemical and biophysical research communications.

[26]  Y. Soini,et al.  Reactive oxygen species and antioxidant mechanisms in human tissues and their relation to malignancies , 2007, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[27]  B. Mannervik,et al.  The amount and nature of glutathione transferases in rat liver microsomes determined by immunochemical methods , 1983, FEBS letters.

[28]  E. Maser,et al.  Peroxide-induced cell death and lipid peroxidation in C6 glioma cells. , 2008, Toxicology in vitro : an international journal published in association with BIBRA.

[29]  Mirko Francesconi,et al.  Overcoming resistance to conventional drugs in Ewing sarcoma and identification of molecular predictors of outcome. , 2009, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[30]  E. Doran,et al.  Mitochondria and cell death. , 2000, Biochemical Society transactions.

[31]  Shaoyu Zhou,et al.  Cumulative and irreversible cardiac mitochondrial dysfunction induced by doxorubicin. , 2001, Cancer research.

[32]  M. Leng,et al.  Interstrand cross-links of cisplatin induce striking distortions in DNA. , 1999, Journal of inorganic biochemistry.

[33]  J. Turrens,et al.  Mitochondrial formation of reactive oxygen species , 2003, The Journal of physiology.

[34]  K. Tew,et al.  Oxidative stress induced in pathologies: the role of antioxidants. , 1999, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[35]  Y. Awasthi,et al.  Self-regulatory role of 4-hydroxynonenal in signaling for stress-induced programmed cell death. , 2008, Free radical biology & medicine.

[36]  N. Saijo,et al.  Differential gene expression profiles and identification of the genes relevant to clinicopathologic factors in colorectal cancer selected by cDNA array method in combination with principal component analysis. , 2003, International journal of oncology.

[37]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[38]  Hideyuki Yamamoto,et al.  Contribution of liver mitochondrial membrane-bound glutathione transferase to mitochondrial permeability transition pores. , 2009, Toxicology and applied pharmacology.

[39]  W. Shepard,et al.  Crystal structure of a double-stranded DNA containing a cisplatin interstrand cross-link at 1.63 A resolution: hydration at the platinated site. , 1999, Nucleic acids research.

[40]  Hua Liu,et al.  Endoplasmic reticulum stress-associated caspase 12 mediates cisplatin-induced LLC-PK1 cell apoptosis. , 2005, Journal of the American Society of Nephrology : JASN.

[41]  R. Armstrong,et al.  Structure, catalytic mechanism, and evolution of the glutathione transferases. , 1997, Chemical research in toxicology.

[42]  S. Srivastava,et al.  Membrane Association of Glutathione S-Transferase mGSTA4-4, an Enzyme That Metabolizes Lipid Peroxidation Products* , 2002, The Journal of Biological Chemistry.

[43]  R. Morgenstern,et al.  Microsomal glutathione S-transferase. Purification, initial characterization and demonstration that it is not identical to the cytosolic glutathione S-transferases A, B and C. , 2005, European journal of biochemistry.

[44]  Shaoyu Zhou,et al.  Interference with calcium-dependent mitochondrial bioenergetics in cardiac myocytes isolated from doxorubicin-treated rats. , 2001, Toxicology and applied pharmacology.

[45]  J. Custódio,et al.  Cisplatin impairs rat liver mitochondrial functions by inducing changes on membrane ion permeability: prevention by thiol group protecting agents. , 2009, Toxicology.

[46]  M. Rubin,et al.  Profiling and verification of gene expression patterns in normal and malignant human prostate tissues by cDNA microarray analysis. , 2001, Neoplasia.

[47]  Johan Hansson,et al.  Cisplatin Induces Endoplasmic Reticulum Stress and Nucleus-independent Apoptotic Signaling* , 2003, The Journal of Biological Chemistry.

[48]  R. Schlapbach,et al.  Oxidants in mitochondria: from physiology to diseases. , 1995, Biochimica et biophysica acta.

[49]  Y. Aniya,et al.  Novel function of glutathione transferase in rat liver mitochondrial membrane: role for cytochrome c release from mitochondria. , 2008, Toxicology and applied pharmacology.