REDOX REGULATION OF ESTROGEN RECEPTOR ALPHA AND SODIUM HYDROGEN EXCHANGER GENE EXPRESSION BY HYDROGEN PEROXIDE CHUA SUI HUAY BSc (HONOURS) A THESIS SUBMITTED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY NUS GRADUATE SCHOOL FOR INTEGRATIVE SCIENCES AND ENGINEERING NATIONAL UNIVERSITY OF SINGAPORE 2011 ACKNOWLEDGEMENTS I want to express my heartfelt gratitude for my supervisor Associate Professor MarieVeronique Clement, Department of Biochemistry I am thankful that she allowed me to join the lab during my honors year and for continuing my graduate study under her A/P Clement was not just a supervisor, but a mentor who is always willing to listen, to encourage and to share her vast knowledge I would like to thank her for her constant encouragement, patience and faith in my abilities to complete this study I would also like to thank my co-supervisor Dr Alan Kumar for his guidance and support throughout the course of my graduate study in NUS I am grateful for Professor Shazib Pervaiz for being on my thesis advisory committee and for his positive feedback on the direction of my project My warmest thanks to my lab mates in Biochemistry department, for making the long days in lab enjoyable Special thanks to San Min and Jeya who helped me to proofread my thesis I also want to thank Mui Khin for helping me with all the ordering of reagents for my project Thank you NUMI girls for being my great lab mates and friends Thanks for being there to cheer me on and also for being part of my sister gang during my wedding I will certainly miss those days that we out together Finally, my deepest gratitude to my family for their encouragement and support throughout the years, and a special thanks to Zong Hao, who has stood by me through the good and bad times ii SUMMARY There is mounting evidence that implicates reactive oxygen species (ROS) as an important signaling molecule in various physiological processes In contrast to the wealth of information on gene up-regulation by ROS, little attention has been paid to the down-regulation of gene expression by ROS In this thesis, we have studied how two important genes, estrogen receptor alpha (ERα) and sodium hydrogen exchanger (NHE1) were redox-regulated by H2O2 In the first part of the study, the regulation of ERα by oxidative stress induced by the exposure of MCF7 cells to H2O2 was investigated The data supported that ERα protein is down-regulated when exposed to oxidative stress The down-regulation of ERα protein occurs not through proteosomal degradation pathway, but via the decrease in ERα mRNA level We found that Akt, MAPK and caspases were not involved in the down-regulation of ERα by H2O2 Instead, H2O2 inhibition of ERα expression involves an oxidation event that is reversible by the addition of reducing agent, DTT We also demonstrated that 15d-PGJ2 suppressed ERα expression via the production of ROS ERα down-regulation resulted in decreased ERα-responsive gene expression and impaired estrogen signaling These effects could have contributed to the growth arrest observed in H2O2 treated MCF7 cells In the second part of the study, the down-regulation of NHE1 by H2O2 was examined We found that the minimal promoter region required for full transcription activation lies on the 0.15kb of NHE1 promoter This region is also regulated by H2O2 The regulation of NHE1 by H2O2 is similar to the regulation of ERα H2O2 targets NHE1 at the mRNA level in a reversible manner The down-regulation involves an oxidation mechanism which is reversible by reducing agents DTT and BME The drug 15diii PGJ2 was also shown to down-regulate NHE1 promoter activity via the production of ROS Finally, the data suggested that AP2 is not the transcription factor for NHE1 and is not a target in H2O2 mediated down-regulation of NHE1 iv TABLE OF CONTENTS ACKNOWLEDGEMENTS ………………………………………………… ………II SUMMARY………………………………………………………………………….III TABLE OF CONTENTS…………………… …………………………………… V LIST OF TABLES………………………………………………………………… X LIST OF FIGURES……………………………………………… ……………… XI ABBREVIATIONS…………………………………………………………… …XVI PUBLICATIONS AND PRESENTATIONS…………………………………… XIX CHAPTER 1: INTRODUCTION 1.1 FREE RADICALS, REACTIVE SPECIES AND REDOX BALANCE 1.1.1 Reactive oxygen species 1.1.2 The antioxidant system 1.1.3 Hydrogen peroxide as a signaling molecule 1.2 REDOX REGULATION OF GENE EXPRESSION 10 1.2.1 Transcriptional regulation 10 1.2.2 mRNA stability 13 1.2.3 Direct oxidative modification 14 1.2.4 Regulation of protein turnover 16 1.3 ESTROGEN RECEPTOR 17 1.3.1 Estrogen and estrogen receptors: Introduction 17 1.3.2 Estrogen receptor and genomic signaling 20 1.3.3 Membrane and cytoplasmic ER signaling 21 1.3.4 Regulation of estrogen receptors 23 1.3.5 Estrogen and estrogen receptors in human breast cancer 27 v 1.4 THE SODIUM HYDROGEN EXCHANGER 29 1.4.1 pH regulation in the cells 29 1.4.2 Mammalian Na+/H+ exchanger 30 1.4.3 NHE1: Basic structure 31 1.4.4 Physiological and pathological roles of NHE1 33 1.4.5 Regulation of NHE1 37 1.4.6 Regulation of NHE1 gene transcription 39 1.5 AIM OF STUDY 41 CHAPTER 2: MATERIALS AND METHODS 43 2.1 MATERIALS 43 2.1.1 Chemicals and reagents 43 2.1.2 Antibodies 44 2.1.3 Plasmids 44 2.1.4 Cell lines and cell culture 45 2.2 METHODS 47 2.2.1 Treatment of cells with hydrogen peroxide (H2O2) and other compounds………………………………………………………………………47 2.2.2 Morphology studies 47 2.2.3 Crystal violet assay 47 2.2.4 Plasmid transfection 48 2.2.5 Small interfering RNA (siRNA) transfection 48 2.2.6 Protein concentration determination 49 2.2.7 SDS-PAGE and western blot 50 2.2.8 Caspase assay 51 2.2.9 Single and dual luciferase assay 52 2.2.10 Chloramphenicol acetyl transferase (CAT) assay 53 2.2.11 RNA isolation 54 2.2.12 Reverse transcription (RT) and real-time chain polymerase reaction (PCR)……………………………………………………………………………54 2.2.13 Intracellular ROS measurement by CM-H2DCFDA 55 2.2.14 Nuclear-cytoplasmic fractionation 55 2.2.15 Electromobility shift assay (EMSA) 56 vi 2.2.16 Statistical analysis 57 CHAPTER 3A: RESULTS – REDOX REGULATION OF ERα 58 3A.1 H2O2 AND THE EXPRESSION OF ERα PROTEIN 58 3A.1.1 Morphology and growth of MCF7 cells upon exposure to H2O2 58 3A.1.2 Effect of H2O2 on the expression of ER protein 61 3A.1.3 Effects of estrogen on H2O2 down-regulation of ERα 66 3A.1.4 Time-dependent regulation of ERα protein by H2O2 68 3A.1.5 Chronic ROS leads to continuous suppression of ERα 70 3A.1.6 ROS involved in the down-regulation of ERα expression 71 3A.1.7 Effect of peroxynitrite on the expression of ERα protein 73 3A.2 DOWN-REGULATION OF ERα DOES NOT INVOLVE THE PROTEOSOMAL DEGRADATION PATHWAY 74 3A.3 THE DOWN-REGULATION OF ERα BY H2O2 IS VIA DECREASE IN ERα mRNA LEVEL 78 3A.3.1 Effects of H2O2 on the expression of ERα mRNA level 78 3A.3.2 New protein synthesis is not required for H2O2-induced down-regulation of ERα mRNA 81 3A.4 MECHANISM INVOLVED IN H2O2-INDUCED DOWN-REGULATION OF ERα 82 3A.4.1 Role of Akt activation in H2O2 down-regulation of ERα 82 3A.4.2 Role of MAPK activation in H2O2 down-regulation of ERα 84 3A.4.3 Role of caspases in H2O2 down-regulation of ERα 87 3A.4.4 Oxidation is involved in the down-regulation of ERα by H2O2 89 3A.5 EFFECTS OF ERα DOWN-REGULATION ON ER RESPONSE GENES.……………………………………………………………………………94 3A.5.1 Down-regulation of ERα by H2O2 inhibits estrogen response element (ERE) activity 94 3A.5.2 H2O2 down-regulates ERα response genes 96 vii 3A.6 15-DEOXY-DELTA-12,14-PROSTAGLANDIN J2 (15D-PGJ2) DOWNREGULATION OF ERα IS VIA ROS 99 3A.6.1 Morphology and growth of MCF7 cells exposed to 15d-PGJ2 99 3A.6.2 15d-PGJ2 down-regulates ERα at the protein and mRNA level 101 3A.6.3 15d-PGJ2 produces ROS in MCF7 cells 103 3A.6.4 Scavenging of ROS by NAC restores cell growth and ERα expression … ……………………………………………………………… 107 CHAPTER 3B: RESULTS – REDOX REGULATION OF NHE1…………… …110 3B.1 DOWN-REGULATION OF NHE1 BY H2O2 110 3B.1.1 Identification of crucial region in the promoter of NHE1 110 3B.1.2 H2O2 down-regulates 0.15kb proximal NHE1 promoter activity 114 3B.1.3 Oxidation is involved in the down-regulation of 0.15kb proximal NHE1 promoter activity and endogenous mRNA level by H2O2 118 3B.1.4 Peroxynitrite down-regulates 0.15kb NHE1 promoter activity 124 3B.1.5 Role of caspases in H2O2-mediated down-regulation of 0.15kb NHE1 proximal promoter activity 125 3B.1.6 15d-PGJ2 down-regulates 0.15kb NHE1 promoter activity and NHE1 mRNA level via ROS production 129 3B.1.7 H2O2 down-regulates NHE1 expression in MCF7 breast cancer cells ……………………………………………………………………………135 3B.2 TRANSCRIPTION FACTOR INVOLVED IN H2O2 DOWN-REGULATION OF NHE1 EXPRESSION 141 3B.2.1 Analysis of the region between 0.15kb and 0.12kb of NHE1 promoter …………………………………………………………………… 141 3B.2.2 Transcription factor AP2, SP1 and COUP 145 3B.2.3 Investigation of AP2 as transcription factor for NHE1 148 CHAPTER 4: DISCUSSION……………………………………………………….162 4.1 REDOX REGULATION OF ERα 162 4.1.1 Regulation of ERα expression and function by oxidative stress 163 4.1.2 Oxidation as the mechanism mediating H2O2-induced down-regulation of ERα…………………………………………………………………………170 4.1.3 Significances of ERα as a redox regulated gene 172 4.1.4 ROS-induced suppression of ERα gene expression: Possible development of ERα-negative breast cancer phenotype 178 viii 4.2 REDOX REGULATION OF NHE1 181 4.2.1 H2O2 regulates an important region of NHE1 promoter 181 4.2.2 Targeting NHE1 expression through ROS-producing agents ……….184 4.2.3 AP2 is a transcription factor of NHE1: True or false? 186 4.3 ERα AND NHE1: TWO IMPORTANT MEDIATORS OF CANCER CELL SURVIVAL 189 4.3.1 H2O2 regulates ERα and NHE1 in similar ways 189 4.3.2 Relevance of ERα and NHE1 expression as prognosis markers in breast cancer………………………………………………………………………… 191 4.4 FUTURE WORK 192 4.4.1 To determine if H2O2 affect the stability of ERα mRNA 192 4.4.2 To investigate the mechanism of ROS-producing compounds in suppression of ERα expression …193 4.4.3 To identify the transcription factor that regulates NHE1 transcription……………………………………………………………………194 4.5 MODEL AND CONCLUSION 195 APPENDICES 197 REFERENCES 202 ix LIST OF TABLES Table I: Characteristics of H2O2 as second messenger Table 1: Software used in the analysis of the 40 bp of 0.15kb NHE1 promoter 143 Table 2: Transcription factors predicted to bind to the 40 bp nucleotide sequence of 0.15kb NHE1 promoter 144 Table 3: List of compounds and growth factors found to down-regulate ERα and produce ROS 177 Table 4: Similarities between the down-regulation of ERα and NHE1 expression by H2O2……………… 190 x nitrosylated PKB alpha/Akt1 from rat soleus muscle using CapLC-Q-TOF(micro) mass spectrometry J Mass Spectrom 40, 1140-1148 Ma, S., Ochi, H., Cui, L., Zhang, J., and He, W (2003) Hydrogen peroxide induced downregulation of CD28 expression of Jurkat cells is associated with a change of site alphaspecific nuclear factor binding activity and the activation of caspase-3 Exp Gerontol 38, 1109-1118 Mahadev, K., Zilbering, A., Zhu, L., and Goldstein, B (2001) Insulin-stimulated hydrogen peroxide reversibly inhibits protein-tyrosine phosphatase 1b in vivo and enhances the early insulin action cascade J Biol Chem 276, 21938-21942 Mailänder, J., Müller-Esterl, W., and Dedio, J (2001) Human homolog of mouse tescalcin associates with Na(+)/H(+) exchanger type-1 FEBS Lett 507, 331-335 Malakooti, J., Dahdal, R., Schmidt, L., Layden, T., Dudeja, P., and Ramaswamy, K (1999) Molecular cloning, tissue distribution, and functional expression of the human Na(+)/H(+) exchanger NHE2 Am J Physiol 277, G383-G390 Mallis, R J., Buss, J E., and Thomas, J A (2001) Oxidative modification of H-ras Sthiolation and S-nitrosylation of reactive cysteines Biochem J 355, 145-153 Malo, M., and Fliegel, L (2006) Physiological role and regulation of the Na+/H+ exchanger Can J Physiol Pharmacol 84, 1081-1095 Mangelsdorf, D., Thummel, C., Beato, M., Herrlich, P., Schütz, G., Umesono, K., Blumberg, B., Kastner, P., Mark, M., and Chambon, P., et al (1995) The nuclear receptor superfamily: the second decade Cell 83, 835-839 Mannick, J., Hausladen, A., Liu, L., Hess, D., Zeng, M., Miao, Q., Kane, L., Gow, A., and Stamler, J (1999) Fas-induced caspase denitrosylation Science 284, 651-654 Marshall, H., and Stamler, J (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40, 1688-1693 Martin, D., Salinas, M., Fujita, N., Tsuruo, T., and Cuadrado, A (2002) Ceramide and reactive oxygen species generated by H2O2 induce caspase-3-independent degradation of Akt/protein kinase B J Biol Chem 277, 42943-42952 Martin, M., Franke, T., Stoica, G., Chambon, P., Katzenellenbogen, B., Stoica, B., McLemore, M., Olivo, S., and Stoica, A (2000) A role for Akt in mediating the estrogenic functions of epidermal growth factor and insulin-like growth factor I Endocrinology 141, 4503-4511 McClelland, R., Barrow, D., Madden, T., Dutkowski, C., Pamment, J., Knowlden, J., Gee, J., and Nicholson, R (2001) Enhanced epidermal growth factor receptor signaling in MCF7 breast cancer cells after long-term culture in the presence of the pure antiestrogen ICI 182,780 (Faslodex) Endocrinology 142, 2776-2788 McInerney, E., Weis, K., Sun, J., Mosselman, S., and Katzenellenbogen, B (1998) Transcription activation by the human estrogen receptor subtype beta (ER beta) studied with ER beta and ER alpha receptor chimeras Endocrinology 139, 4513-4522 McLean, L., Roscoe, J., Jorgensen, N., Gorin, F., and Cala, P (2000) Malignant gliomas display altered pH regulation by NHE1 compared with nontransformed astrocytes Am J Physiol Cell Physiol 278, C676-C688 220 McPherson, L., Baichwal, V., and Weigel, R (1997) Identification of ERF-1 as a member of the AP2 transcription factor family Proc Natl Acad Sci U S A 94, 4342-4347 McPherson, L., and Weigel, R (1999) AP2alpha and AP2gamma: a comparison of binding site specificity and trans-activation of the estrogen receptor promoter and single site promoter constructs Nucleic Acids Res 27, 4040-4049 Meshnick, S R., Yang, Y Z., Lima, V., Kuypers, F., Kamchonwongpaisan, S., and Yuthavong, Y (1993) Iron-dependent free radical generation from the antimalarial agent artemisinin qinghaosu Antimicrob Agents Chemother 37, 1108-1114 Miller, R., Counillon, L., Pages, G., Lifton, R., Sardet, C., and Pouysségur, J (1991) Structure of the 5'-flanking regulatory region and gene for the human growth factoractivatable Na/H exchanger NHE-1 J Biol Chem 266, 10813-10819 Minicis, S D., and Brenner, D A (2007) NOX in liver fibrosis Arch Biochem Biophys 462, 266-272 Misik, A., Perreault, K., Holmes, C., and Fliegel, L (2005) Protein phosphatase regulation of Na+/H+ exchanger isoform I Biochemistry 44, 5842-5852 Mitchell, P., Wang, C., and Tjian, R (1987) Positive and negative regulation of transcription in vitro: enhancer-binding protein AP-2 is inhibited by SV40 T antigen Cell 50, 847-861 Mitomo, K., Nakayama, K., Fujimoto, K., Sun, X., Seki, S., and Yamamoto, K (1994) Two different cellular redox systems regulate the DNA-binding activity of the p50 subunit of NFkappa B in vitro Gene 145, 197-203 Mize, A., Shapiro, R., and Dorsa, D (2003) Estrogen receptor-mediated neuroprotection from oxidative stress requires activation of the mitogen-activated protein kinase pathway Endocrinology 144, 306-312 Mohibullah, N., Donner, A., Ippolito, J., and Williams, T (1999) SELEX and missing phosphate contact analyses reveal flexibility within the AP-2[alpha] protein: DNA binding complex Nucleic Acids Res 27, 2760-2769 Montano, M., Müller, V., Trobaugh, A., and Katzenellenbogen, B (1995) The carboxyterminal F domain of the human estrogen receptor: role in the transcriptional activity of the receptor and the effectiveness of antiestrogens as estrogen antagonists Mol Endocrinol 9, 814-825 Morel, Y., and Barouki, R (1998) Down-regulation of cytochrome P450 1A1 gene promoter by oxidative stress Critical contribution of nuclear factor J Biol Chem 273, 26969-26976 Morel, Y., and Barouki, R (2000) The repression of nuclear factor ICCAAT transcription factor NFICTF transactivating domain by oxidative stress is mediated by a critical cysteine Cys-427 Biochem J 348, 235-240 Mosselman, S., Polman, J., and Dijkema, R (1996) ER beta: identification and characterization of a novel human estrogen receptor FEBS Lett 392, 49-53 Motohashi, H., Katsuoka, F., Engel, J., and Yamamoto, M (2004) Small Maf proteins serve as transcriptional cofactors for keratinocyte differentiation in the Keap1-Nrf2 regulatory pathway Proc Natl Acad Sci U S A 101, 6379-6384 221 Murphy, G., and Holder, J (2000) PPAR-gamma agonists: therapeutic role in diabetes, inflammation and cancer Trends Pharmacol Sci 21, 469-474 Nadal, A., Díaz, M., and Valverde, M (2001) The estrogen trinity: membrane, cytosolic, and nuclear effects News Physiol Sci 16, 251-255 Nakamura, K., Hori, T., Sato, N., Sugie, K., Kawakami, T., and Yodoi, J (1993) Redox regulation of a src family protein tyrosine kinase p56lck in T cells Oncogene 8, 3133-3139 Nakamura, N., Tanaka, S., Teko, Y., Mitsui, K., and Kanazawa, H (2005) Four Na+/H+ exchanger isoforms are distributed to Golgi and post-Golgi compartments and are involved in organelle pH regulation J Biol Chem 280, 1561-1572 Nakazato, T., Ito, K., Miyakawa, Y., Kinjo, K., Yamada, T., Hozumi, N., Ikeda, Y., and Kizaki, M (2005) Catechin, a green tea component, rapidly induces apoptosis of myeloid leukemic cells via modulation of reactive oxygen species production in vitro and inhibits tumor growth in vivo Haematologica 90, 317-325 Nauseef, W (2004) Assembly of the phagocyte NADPH oxidase Histochem Cell Biol 122, 277-291 Nawaz, Z., Lonard, D M., Dennis, A P., Smith, C L., and OMalley, B W (1999) Proteasome-dependent degradation of the human estrogen receptor Proc Natl Acad Sci U S A 96, 1858-1862 Newman, S., Bates, N., Vernimmen, D., Parker, M., and Hurst, H (2000) Cofactor competition between the ligand-bound oestrogen receptor and an intron enhancer leads to oestrogen repression of ERBB2 expression in breast cancer Oncogene 19, 490-497 Nikinmaa, M., Pursiheimo, S., and Soitamo, A (2004) Redox state regulates HIF-1alpha and its DNA binding and phosphorylation in salmonid cells J Cell Sci 117, 3201-3206 Nikitakis, N G., Siavash, H., Hebert, C., Reynolds, M A., Hamburger, A W., and Sauk, J J (2002) 15-PGJ2 but not thiazolidinediones inhibits cell growth induces apoptosis and causes downregulation of Stat3 in human oral SCCa cells Br J Cancer 87, 1396-1403 Nilsson, S., Mäkelä, S., Treuter, E., Tujague, M., Thomsen, J., Andersson, G., Enmark, E., Pettersson, K., Warner, M., and Gustafsson, J (2001) Mechanisms of estrogen action Physiol Rev 81, 1535-1565 Nirmala, P., and Thampan, R (1995) Ubiquitination of the rat uterine estrogen receptor: dependence on estradiol Biochem Biophys Res Commun 213, 24-31 Nutter, L., Wu, Y., Ngo, E., Sierra, E., Gutierrez, P., and Abul-Hajj, Y (1994) An o-quinone form of estrogen produces free radicals in human breast cancer cells: correlation with DNA damage Chem Res Toxicol 7, 23-28 O'Brian, C., and Chu, F (2005) Post-translational disulfide modifications in cell signaling-role of inter-protein, intra-protein, S-glutathionyl, and S-cysteaminyl disulfide modifications in signal transmission Free Radic Res 39, 471-480 Ober, S., and Pardee, A (1987) Intracellular pH is increased after transformation of Chinese hamster embryo fibroblasts Proc Natl Acad Sci U S A 84, 2766-2770 Ogawa, S., Inoue, S., Watanabe, T., Hiroi, H., Orimo, A., Hosoi, T., Ouchi, Y., and 222 Muramatsu, M (1998) The complete primary structure of human estrogen receptor beta (hER beta) and its heterodimerization with ER alpha in vivo and in vitro Biochem Biophys Res Commun 243, 122-126 Oh, A., Lorant, L., Holloway, J., Miller, D., Kern, F., and El-Ashry, D (2001) Hyperactivation of MAPK induces loss of ERalpha expression in breast cancer cells Mol Endocrinol 15, 1344-1359 Orlowski, J., and Grinstein, S (2004) Diversity of the mammalian sodium/proton exchanger SLC9 gene family Pflugers Arch 447, 549-565 Orlowski, J., Kandasamy, R., and Shull, G (1992) Molecular cloning of putative members of the Na/H exchanger gene family cDNA cloning, deduced amino acid sequence, and mRNA tissue expression of the rat Na/H exchanger NHE-1 and two structurally related proteins J Biol Chem 267, 9331-9339 Osborne, C., Wilson, P., and Tripathy, D (2004) Oncogenes and tumor suppressor genes in breast cancer: potential diagnostic and therapeutic applications Oncologist 9, 361-377 Ottaviano, Y., Issa, J., Parl, F., Smith, H., Baylin, S., and Davidson, N (1994) Methylation of the estrogen receptor gene CpG island marks loss of estrogen receptor expression in human breast cancer cells Cancer Res 54, 2552-2555 Paech, K., Webb, P., Kuiper, G., Nilsson, S., Gustafsson, J., Kushner, P., and Scanlan, T (1997) Differential ligand activation of estrogen receptors ERalpha and ERbeta at AP1 sites Science 277, 1508-1510 Pakdel, F., Le, G., and Katzenellenbogen, B (1993) An assessment of the role of domain F and PEST sequences in estrogen receptor half-life and bioactivity J Steroid Biochem Mol Biol 46, 663-672 Pang, T., Hisamitsu, T., Mori, H., Shigekawa, M., and Wakabayashi, S (2004) Role of calcineurin B homologous protein in pH regulation by the Na+/H+ exchanger 1: tightly bound Ca2+ ions as important structural elements Biochemistry 43, 3628-3636 Pantano, C., Anathy, V., Ranjan, P., Heintz, N., and Janssen-Heininger, Y (2007) Nonphagocytic oxidase causes death in lung epithelial cells via a TNF-RI-JNK signaling axis Am J Respir Cell Mol Biol 36, 473-479 Paruthiyil, S., Parmar, H., Kerekatte, V., Cunha, G., Firestone, G., and Leitman, D (2004) Estrogen receptor beta inhibits human breast cancer cell proliferation and tumor formation by causing a G2 cell cycle arrest Cancer Res 64, 423-428 Pearce, S., and Jordan, V (2004) The biological role of estrogen receptors alpha and beta in cancer Crit Rev Oncol Hematol 50, 3-22 Pedram, A., Razandi, M., and Levin, E (2006) Nature of functional estrogen receptors at the plasma membrane Mol Endocrinol 20, 1996-2009 Pelicano, H., Feng, L., Zhou, Y., Carew, J., Hileman, E., Plunkett, W., Keating, M., and Huang, P (2003) Inhibition of mitochondrial respiration: a novel strategy to enhance druginduced apoptosis in human leukemia cells by a reactive oxygen species-mediated mechanism J Biol Chem 278, 37832-37839 223 Pervaiz, S., and Clement, M (2004) Tumor intracellular redox status and drug resistance-serendipity or a causal relationship? Curr Pharm Des 10, 1969-1977 Pervaiz, S., and Clément, M (2002) A permissive apoptotic environment: function of a decrease in intracellular superoxide anion and cytosolic acidification Biochem Biophys Res Commun 290, 1145-1150 Petrecca, K., Atanasiu, R., Grinstein, S., Orlowski, J., and Shrier, A (1999) Subcellular localization of the Na+/H+ exchanger NHE1 in rat myocardium Am J Physiol 276, H709H717 Pettersson, K., and Gustafsson, J (2001) Role of estrogen receptor beta in estrogen action Annu Rev Physiol 63, 165-192 Pickart, C (1997) Targeting of substrates to the 26S proteasome FASEB J 11, 1055-1066 Pietras, R., and Szego, C (1977) Specific binding sites for oestrogen at the outer surfaces of isolated endometrial cells Nature 265, 69-72 Pineda-Molina, E., Klatt, P., Vázquez, J., Marina, A., García, D., Pérez-Sala, D., and Lamas, S (2001) Glutathionylation of the p50 subunit of NF-kappaB: a mechanism for redoxinduced inhibition of DNA binding Biochemistry 40, 14134-14142 Pinzone, J J., Stevenson, H., Strobl, J S., and Berg, P E (2004) Molecular and Cellular Determinants of Estrogen Receptor Expression Mol Cell Biol 24, 4605-4612 Poole, L., Karplus, P., and Claiborne, A (2004) Protein sulfenic acids in redox signaling Annu Rev Pharmacol Toxicol 44, 325-347 Poole, L B., and Nelson, K J (2008) Discovering mechanisms of signaling-mediated cysteine oxidation Curr Opin Chem Biol 12, 18-24 Pouyssgur, J., Chambard, J C., Franchi, A., Paris, S., and Obberghen-Schilling, E V (1982) Growth factor activation of an amiloride-sensitive NaH exchange system in quiescent fibroblasts coupling to ribosomal protein S6 phosphorylation Proc Natl Acad Sci U S A 79, 3935-3939 Pouysségur, J., Sardet, C., Franchi, A., L'Allemain, G., and Paris, S (1984) A specific mutation abolishing Na+/H+ antiport activity in hamster fibroblasts precludes growth at neutral and acidic pH Proc Natl Acad Sci U S A 81, 4833-4837 Pozo-Guisado, E., Merino, J., Mulero-Navarro, S., Lorenzo-Benayas, M., Centeno, F., Alvarez-Barrientos, A., and Fernandez-Salguero, P (2005) Resveratrol-induced apoptosis in MCF-7 human breast cancer cells involves a caspase-independent mechanism with downregulation of Bcl-2 and NF-kappaB Int J Cancer 115, 74-84 Prall, O., Sarcevic, B., Musgrove, E., Watts, C., and Sutherland, R (1997) Estrogen-induced activation of Cdk4 and Cdk2 during G1-S phase progression is accompanied by increased cyclin D1 expression and decreased cyclin-dependent kinase inhibitor association with cyclin E-Cdk2 J Biol Chem 272, 10882-10894 Pryzbylkowski, P., Obajimi, O., and Keen, J (2008) Trichostatin A and Aza-2' deoxycytidine decrease estrogen receptor mRNA stability in ER positive MCF7 cells through modulation of HuR Breast Cancer Res Treat 111, 15-25 224 Putney, L., and Barber, D (2003) Na-H exchange-dependent increase in intracellular pH times G2/M entry and transition J Biol Chem 278, 44645-44649 Putney, L., Denker, S., and Barber, D (2002) The changing face of the Na+/H+ exchanger, NHE1: structure, regulation, and cellular actions Annu Rev Pharmacol Toxicol 42, 527-552 Pérez-Mato, I., Castro, C., Ruiz, F., Corrales, F., and Mato, J (1999) Methionine adenosyltransferase S-nitrosylation is regulated by the basic and acidic amino acids surrounding the target thiol J Biol Chem 274, 17075-17079 Qin, C., Burghardt, R., Smith, R., Wormke, M., Stewart, J., and Safe, S (2003) Peroxisome proliferator-activated receptor gamma agonists induce proteasome-dependent degradation of cyclin D1 and estrogen receptor alpha in MCF-7 breast cancer cells Cancer Res 63, 958-964 Rahman, I (2003) Oxidative stress, chromatin remodeling and gene transcription in inflammation and chronic lung diseases J Biochem Mol Biol 36, 95-109 Rahman, I., Gilmour, P., Jimenez, L., and MacNee, W (2002) Oxidative stress and TNFalpha induce histone acetylation and NF-kappaB/AP-1 activation in alveolar epithelial cells: potential mechanism in gene transcription in lung inflammation Mol Cell Biochem 235, 239248 Razandi, M., Alton, G., Pem, A., Ghonshani, S., Webb, P., and Levin, E R (2003) Identification of a Structural Determinant Necessary for the Localization and Function of Estrogen Receptor at the Plasma Membrane Mol Cell Biol 23, 1633-1646 Razandi, M., Pedram, A., Park, S., and Levin, E (2003) Proximal events in signaling by plasma membrane estrogen receptors J Biol Chem 278, 2701-2712 Reshkin, S., Bellizzi, A., Caldeira, S., Albarani, V., Malanchi, I., Poignee, M., AlunniFabbroni, M., Casavola, V., and Tommasino, M (2000) Na+/H+ exchanger-dependent intracellular alkalinization is an early event in malignant transformation and plays an essential role in the development of subsequent transformation-associated phenotypes FASEB J 14, 2185-2197 Rhee, S (2006) Cell signaling H2O2, a necessary evil for cell signaling Science 312, 18821883 Rhee, S., Kang, S., Jeong, W., Chang, T., Yang, K., and Woo, H (2005) Intracellular messenger function of hydrogen peroxide and its regulation by peroxiredoxins Curr Opin Cell Biol 17, 183-189 Rich, I., Worthington-White, D., Garden, O., and Musk, P (2000) Apoptosis of leukemic cells accompanies reduction in intracellular pH after targeted inhibition of the Na(+)/H(+) exchanger Blood 95, 1427-1434 Richard, D., Berra, E., and Pouyssegur, J (2000) Nonhypoxic pathway mediates the induction of hypoxia-inducible factor 1alpha in vascular smooth muscle cells J Biol Chem 275, 26765-26771 Rieder, C., and Fliegel, L (2003) Transcriptional regulation of Na+/H+ exchanger expression in the intact mouse Mol Cell Biochem 243, 87-95 Risse, G., Jooss, K., Neuberg, M., Brller, H J., and Mller, R (1989) Asymmetrical recognition of the palindromic AP1 binding site TRE by Fos protein complexes EMBO J 8, 225 3825-3832 Rogatsky, I., Trowbridge, J., and Garabedian, M (1999) Potentiation of human estrogen receptor alpha transcriptional activation through phosphorylation of serines 104 and 106 by the cyclin A-CDK2 complex J Biol Chem 274, 22296-22302 Rogers, S., Wells, R., and Rechsteiner, M (1986) Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis Science 234, 364-368 Roquemore, E., Chevrier, M., Cotter, R., and Hart, G (1996) Dynamic O-GlcNAcylation of the small heat shock protein alpha B-crystallin Biochemistry 35, 3578-3586 Rosenfeld, C., White, R., Roy, T., and Cox, B (2000) Calcium-activated potassium channels and nitric oxide coregulate estrogen-induced vasodilation Am J Physiol Heart Circ Physiol 279, H319-H328 Ross, J (1995) mRNA stability in mammalian cells Microbiol Rev 59, 423-450 Roveri, A., Coassin, M., Maiorino, M., Zamburlini, A., van, A., Ratti, E., and Ursini, F (1992) Effect of hydrogen peroxide on calcium homeostasis in smooth muscle cells Arch Biochem Biophys 297, 265-270 Russo, J., Hasan, L., Balogh, G., Guo, S., and Russo, I (2003) Estrogen and its metabolites are carcinogenic agents in human breast epithelial cells J Steroid Biochem Mol Biol 87, 1-25 Saceda, M., Lindsey, R K., Solomon, H., Angeloni, S V., and Martin, M B (1998) Estradiol regulates estrogen receptor mRNA stability J Steroid Biochem Mol Biol 66, 113120 Samikkannu, T., Chen, C., Yih, L., Wang, A., Lin, S., Chen, T., and Jan, K (2003) Reactive oxygen species are involved in arsenic trioxide inhibition of pyruvate dehydrogenase activity Chem Res Toxicol 16, 409-414 Sandelin, A., Alkema, W., Engström, P., Wasserman, W., and Lenhard, B (2004) JASPAR: an open-access database for eukaryotic transcription factor binding profiles Nucleic Acids Res 32, D91-D94 Sandelin, A., Wasserman, W., and Lenhard, B (2004) ConSite: web-based prediction of regulatory elements using cross-species comparison Nucleic Acids Res 32, W249-W252 Sardet, C., Franchi, A., and Pouysségur, J (1989) Molecular cloning, primary structure, and expression of the human growth factor-activatable Na+/H+ antiporter Cell 56, 271-280 Sauer, H., Wartenberg, M., and Hescheler, J (2001) Reactive oxygen species as intracellular messengers during cell growth and differentiation Cell Physiol Biochem 11, 173-186 Sauvage, M., Mazière, P., Fathallah, H., and Giraud, F (2000) Insulin stimulates NHE1 activity by sequential activation of phosphatidylinositol 3-kinase and protein kinase C zeta in human erythrocytes Eur J Biochem 267, 955-962 Savitsky, P., and Finkel, T (2002) Redox regulation of Cdc25C J Biol Chem 277, 2053520540 Schieven, G., Kirihara, J., Burg, D., Geahlen, R., and Ledbetter, J (1993) p72syk tyrosine 226 kinase is activated by oxidizing conditions that induce lymphocyte tyrosine phosphorylation and Ca2+ signals J Biol Chem 268, 16688-16692 Schieven, G., Kirihara, J., Myers, D., Ledbetter, J., and Uckun, F (1993) Reactive oxygen intermediates activate NF-kappa B in a tyrosine kinase-dependent mechanism and in combination with vanadate activate the p56lck and p59fyn tyrosine kinases in human lymphocytes Blood 82, 1212-1220 Schmuck, G., Roehrdanz, E., Haynes, R., and Kahl, R (2002) Neurotoxic mode of action of artemisinin Antimicrob Agents Chemother 46, 821-827 Schneider, C., Tallman, K., Porter, N., and Brash, A (2001) Two distinct pathways of formation of 4-hydroxynonenal Mechanisms of nonenzymatic transformation of the 9- and 13-hydroperoxides of linoleic acid to 4-hydroxyalkenals J Biol Chem 276, 20831-20838 Scholz, W., Albus, U., Counillon, L., Gögelein, H., Lang, H., Linz, W., Weichert, A., and Schölkens, B (1995) Protective effects of HOE642, a selective sodium-hydrogen exchange subtype inhibitor, on cardiac ischaemia and reperfusion Cardiovasc Res 29, 260-268 Schug, J (2008) Using TESS to predict transcription factor binding sites in DNA sequence Curr Protoc Bioinformatics Schumacker, P (2006) Reactive oxygen species in cancer cells: live by the sword, die by the sword Cancer Cell 10, 175-176 Schuur, E R., McPherson, L A., Yang, G P., and Weigel, R J (2001) Genomic structure of the promoters of the human estrogen receptor-alpha gene demonstrate changes in chromatin structure induced by AP2gamma J Biol Chem 276, 15519-15526 Scott, G K., Kushner, P., Vigne, J L., and Benz, C C (1991) Truncated forms of DNAbinding estrogen receptors in human breast cancer J Clin Invest 88, 700-706 Seacat, A., Kuppusamy, P., Zweier, J., and Yager, J (1997) ESR identification of free radicals formed from the oxidation of catechol estrogens by Cu2+ Arch Biochem Biophys 347, 45-52 Seargent, J., Yates, E., and Gill, J (2004) GW9662, a potent antagonist of PPARgamma, inhibits growth of breast tumour cells and promotes the anticancer effects of the PPARgamma agonist rosiglitazone, independently of PPARgamma activation Br J Pharmacol 143, 933937 Seargent, J M., Yates, E A., and Gill, J H (2004) GW9662 a potent antagonist of PPAR inhibits growth of breast tumour cells and promotes the anticancer effects of the PPAR agonist rosiglitazone independently of PPAR activation Br J Pharmacol 143, 933-937 Sen, C., and Packer, L (1996) Antioxidant and redox regulation of gene transcription FASEB J 10, 709-720 Senoo-Matsuda, N., Yasuda, K., Tsuda, M., Ohkubo, T., Yoshimura, S., Nakazawa, H., Hartman, P., and Ishii, N (2001) A defect in the cytochrome b large subunit in complex II causes both superoxide anion overproduction and abnormal energy metabolism in Caenorhabditis elegans J Biol Chem 276, 41553-41558 Shaulian, E., and Karin, M (2002) AP-1 as a regulator of cell life and death Nat Cell Biol 4, E131-E136 227 Shi, M., Godleski, J., and Paulauskis, J (1996) Regulation of macrophage inflammatory protein-1alpha mRNA by oxidative stress J Biol Chem 271, 5878-5883 Shin, S W., Seo, C Y., Han, H., Han, J Y., Jeong, J S., Kwak, J Y., and Park, J I (2009) 15d-PGJ2 induces apoptosis by reactive oxygen species-mediated inactivation of Akt in leukemia and colorectal cancer cells and shows in vivo antitumor activity Clin Cancer Res 15, 5414-5425 Siczkowski, M., Davies, J., and Ng, L (1994) Activity and density of the Na+/H+ antiporter in normal and transformed human lymphocytes and fibroblasts Am J Physiol 267, C745C752 Silva, N., Haworth, R., Singh, D., and Fliegel, L (1995) The carboxyl-terminal region of the Na+/H+ exchanger interacts with mammalian heat shock protein Biochemistry 34, 1041210420 Silverman, E., and Drazen, J (2000) Genetic variations in the 5-lipoxygenase core promoter Description and functional implications Am J Respir Crit Care Med 161, S77-S80 Simpson, E., and Davis, S (2001) Minireview: aromatase and the regulation of estrogen biosynthesis some new perspectives Endocrinology 142, 4589-4594 Smith, A., and Humphries, S (2009) Characterization of DNA-binding proteins using multiplexed competitor EMSA J Mol Biol 385, 714-717 Snabaitis, A., D'Mello, R., Dashnyam, S., and Avkiran, M (2006) A novel role for protein phosphatase 2A in receptor-mediated regulation of the cardiac sarcolemmal Na+/H+ exchanger NHE1 J Biol Chem 281, 20252-20262 Song, R., McPherson, R., Adam, L., Bao, Y., Shupnik, M., Kumar, R., and Santen, R (2002) Linkage of rapid estrogen action to MAPK activation by ERalpha-Shc association and Shc pathway activation Mol Endocrinol 16, 116-127 Soule, H., Vazguez, J., Long, A., Albert, S., and Brennan, M (1973) A human cell line from a pleural effusion derived from a breast carcinoma J Natl Cancer Inst 51, 1409-1416 Soum, E., Brazzolotto, X., Goussias, C., Bouton, C., Moulis, J., Mattioli, T., and Drapier, J (2003) Peroxynitrite and nitric oxide differently target the iron-sulfur cluster and amino acid residues of human iron regulatory protein Biochemistry 42, 7648-7654 Stadtman, E., and Berlett, B (1991) Fenton chemistry Amino acid oxidation J Biol Chem 266, 17201-17211 Stefano, G., Prevot, V., Beauvillain, J., Cadet, P., Fimiani, C., Welters, I., Fricchione, G., Breton, C., Lassalle, P., and Salzet, M., et al (2000) Cell-surface estrogen receptors mediate calcium-dependent nitric oxide release in human endothelia Circulation 101, 1594-1597 Stinefelt, B., Leonard, S., Blemings, K., Shi, X., and Klandorf, H (2005) Free radical scavenging, DNA protection, and inhibition of lipid peroxidation mediated by uric acid Ann Clin Lab Sci 35, 37-45 Stoica, A., Saceda, M., Doraiswamy, V., Coleman, C., and Martin, M (2000) Regulation of estrogen receptor-alpha gene expression by epidermal growth factor J Endocrinol 165, 371378 228 Stoner, M., Saville, B., Wormke, M., Dean, D., Burghardt, R., and Safe, S (2001) Hypoxia Induces Proteasome-Dependent Degradation of Estrogen Receptor in ZR-75 Breast Cancer Cells Molecular Endocrinology 16, 2001-0347 Ström, A., Hartman, J., Foster, J., Kietz, S., Wimalasena, J., and Gustafsson, J (2004) Estrogen receptor beta inhibits 17beta-estradiol-stimulated proliferation of the breast cancer cell line T47D Proc Natl Acad Sci U S A 101, 1566-1571 Suc, I., Meilhac, O., Lajoie-Mazenc, I., Vandaele, J., Jürgens, G., Salvayre, R., and NègreSalvayre, A (1998) Activation of EGF receptor by oxidized LDL FASEB J 12, 665-671 Sumimoto, H., Ueno, N., Yamasaki, T., Taura, M., and Takeya, R (2004) Molecular mechanism underlying activation of superoxide-producing NADPH oxidases: roles for their regulatory proteins Jpn J Infect Dis 57, S24-S25 Sun, G., Porter, W., and Safe, S (1998) Estrogen-induced retinoic acid receptor alpha gene expression: role of estrogen receptor-Sp1 complex Mol Endocrinol 12, 882-890 Sun, J., Steenbergen, C., and Murphy, E (2006) S-nitrosylation: NO-related redox signaling to protect against oxidative stress Antioxid Redox Signal 8, 1693-1705 Sundar, S N., Marconett, C N., Doan, V B., Willoughby, J A., Sr, and Firestone, G L (2008) Artemisinin selectively decreases functional levels of estrogen receptor-alpha and ablates estrogen-induced proliferation in human breast cancer cells Carcinogenesis 29, 22522258 Sundaresan, M., Yu, Z., Ferrans, V., Irani, K., and Finkel, T (1995) Requirement for generation of H2O2 for platelet-derived growth factor signal transduction Science 270, 296299 Suske, G., Bruford, E., and Philipsen, S (2005) Mammalian SP/KLF transcription factors: bring in the family Genomics 85, 551-556 Swami, S., Krishnan, A., and Feldman, D (2000) 1alpha,25-Dihydroxyvitamin D3 downregulates estrogen receptor abundance and suppresses estrogen actions in MCF-7 human breast cancer cells Clin Cancer Res 6, 3371-3379 Szego, C., and Davis, J (1967) Adenosine 3',5'-monophosphate in rat uterus: acute elevation by estrogen Proc Natl Acad Sci U S A 58, 1711-1718 Sánchez, Y., Amrán, D., de, B., and Aller, P (2009) Regulation of genistein-induced differentiation in human acute myeloid leukaemia cells (HL60, NB4) Protein kinase modulation and reactive oxygen species generation Biochem Pharmacol 77, 384-396 Sørensen, V., Zhen, Y., Zakrzewska, M., Haugsten, E., Wälchli, S., Nilsen, T., Olsnes, S., and Wiedlocha, A (2008) Phosphorylation of fibroblast growth factor (FGF) receptor at Ser777 by p38 mitogen-activated protein kinase regulates translocation of exogenous FGF1 to the cytosol and nucleus Mol Cell Biol 28, 4129-4141 Takahashi, E., Abe, J., Gallis, B., Aebersold, R., Spring, D., Krebs, E., and Berk, B (1999) p90(RSK) is a serum-stimulated Na+/H+ exchanger isoform-1 kinase Regulatory phosphorylation of serine 703 of Na+/H+ exchanger isoform-1 J Biol Chem 274, 2020620214 229 Takeya, R., Ueno, N., and Sumimoto, H (2006) Regulation of superoxide-producing NADPH oxidases in nonphagocytic cells Methods Enzymol 406, 456-468 Tamir, S., Izrael, S., and Vaya, J (2002) The effect of oxidative stress on ERalpha and ERbeta expression J Steroid Biochem Mol Biol 81, 327-332 Tanito, M., Agbaga, M., and Anderson, R (2007) Upregulation of thioredoxin system via Nrf2-antioxidant responsive element pathway in adaptive-retinal neuroprotection in vivo and in vitro Free Radic Biol Med 42, 1838-1850 Tapryal, N., Mukhopadhyay, C., Das, D., Fox, P., and Mukhopadhyay, C (2009) Reactive oxygen species regulate ceruloplasmin by a novel mRNA decay mechanism involving its 3'untranslated region: implications in neurodegenerative diseases J Biol Chem 284, 1873-1883 Tateishi, Y., Kawabe, Y., Chiba, T., Murata, S., Ichikawa, K., Murayama, A., Tanaka, K., Baba, T., Kato, S., and Yanagisawa, J (2004) Ligand-dependent switching of ubiquitinproteasome pathways for estrogen receptor EMBO J 23, 4813-4823 Teixeira, C., Reed, J., and Pratt, M (1995) Estrogen promotes chemotherapeutic drug resistance by a mechanism involving Bcl-2 proto-oncogene expression in human breast cancer cells Cancer Res 55, 3902-3907 Toledano, M., and Leonard, W (1991) Modulation of transcription factor NF-kappa B binding activity by oxidation-reduction in vitro Proc Natl Acad Sci U S A 88, 4328-4332 Tolhurst, R., Thomas, R., Kyle, F., Patel, H., Periyasamy, M., Photiou, A., Thiruchelvam, P., Lai, C., Al-Sabbagh, M., and Fisher, R., et al (2010) Transient over-expression of estrogen receptor-alpha in breast cancer cells promotes cell survival and estrogen-independent growth Breast Cancer Res Treat 128, 357-368 Tominaga, T., and Barber, D (1998) Na-H exchange acts downstream of RhoA to regulate integrin-induced cell adhesion and spreading Mol Biol Cell 9, 2287-2303 Tominaga, T., Ishizaki, T., Narumiya, S., and Barber, D (1998) p160ROCK mediates RhoA activation of Na-H exchange EMBO J 17, 4712-4722 Tonks, N (2005) Redox redux: revisiting PTPs and the control of cell signaling Cell 121, 667-670 Toualbi, K., Güller, M., Mauriz, J., Labalette, C., Buendia, M., Mauviel, A., and Bernuau, D (2007) Physical and functional cooperation between AP-1 and beta-catenin for the regulation of TCF-dependent genes Oncogene 26, 3492-3502 Trachootham, D., Lu, W., Ogasawara, M A., Valle, N R D., and Huang, P (2008) Redox Regulation of Cell Survival Antioxid Redox Signal 10, 1343-1374 Treeck, O., Lattrich, C., Springwald, A., and Ortmann, O (2010) Estrogen receptor beta exerts growth-inhibitory effects on human mammary epithelial cells Breast Cancer Res Treat 120, 557-565 Tremblay, A., Tremblay, G., Labrie, F., and Giguère, V (1999) Ligand-independent recruitment of SRC-1 to estrogen receptor beta through phosphorylation of activation function AF-1 Mol Cell 3, 513-519 Tremblay, G., Tremblay, A., Copeland, N., Gilbert, D., Jenkins, N., Labrie, F., and Giguère, 230 V (1997) Cloning, chromosomal localization, and functional analysis of the murine estrogen receptor beta Mol Endocrinol 11, 353-365 Trumpower, B (1990) The protonmotive Q cycle Energy transduction by coupling of proton translocation to electron transfer by the cytochrome bc1 complex J Biol Chem 265, 1140911412 Tsai, M., and O'Malley, B (1994) Molecular mechanisms of action of steroid/thyroid receptor superfamily members Annu Rev Biochem 63, 451-486 Tsai, S., and Tsai, M (1997) Chick ovalbumin upstream promoter-transcription factors (COUP-TFs): coming of age Endocr Rev 18, 229-240 Turpaev, K (2002) Reactive oxygen species and regulation of gene expression Biochemistry (Mosc) 67, 281-292 Turrens, J (2003) Mitochondrial formation of reactive oxygen species J Physiol 552, 335344 Turrens, J., Alexandre, A., and Lehninger, A (1985) Ubisemiquinone is the electron donor for superoxide formation by complex III of heart mitochondria Arch Biochem Biophys 237, 408-414 Turrens, J., and Boveris, A (1980) Generation of superoxide anion by the NADH dehydrogenase of bovine heart mitochondria Biochem J 191, 421-427 Tzukerman, M., Esty, A., Santiso-Mere, D., Danielian, P., Parker, M., Stein, R., Pike, J., and McDonnell, D (1994) Human estrogen receptor transactivational capacity is determined by both cellular and promoter context and mediated by two functionally distinct intramolecular regions Mol Endocrinol 8, 21-30 Uchida, K., Shiraishi, M., Naito, Y., Torii, Y., Nakamura, Y., and Osawa, T (1999) Activation of stress signaling pathways by the end product of lipid peroxidation 4-hydroxy-2nonenal is a potential inducer of intracellular peroxide production J Biol Chem 274, 22342242 Urata, Y., Ihara, Y., Murata, H., Goto, S., Koji, T., Yodoi, J., Inoue, S., and Kondo, T (2006) 17Beta-estradiol protects against oxidative stress-induced cell death through the glutathione/glutaredoxin-dependent redox regulation of Akt in myocardiac H9c2 cells J Biol Chem 281, 13092-13102 Ushio-Fukai, M., Alexander, R., Akers, M., and Griendling, K (1998) p38 Mitogen-activated protein kinase is a critical component of the redox-sensitive signaling pathways activated by angiotensin II Role in vascular smooth muscle cell hypertrophy J Biol Chem 273, 1502215029 Vaheri, A., Carpén, O., Heiska, L., Helander, T., Jääskeläinen, J., Majander-Nordenswan, P., Sainio, M., Timonen, T., and Turunen, O (1997) The ezrin protein family: membranecytoskeleton interactions and disease associations Curr Opin Cell Biol 9, 659-666 Valks, D., Kemp, T., and Clerk, A (2003) Regulation of Bcl-xL expression by H2O2 in cardiac myocytes J Biol Chem 278, 25542-25547 Veal, E., Day, A., and Morgan, B (2007) Hydrogen peroxide sensing and signaling Mol Cell 26, 1-14 231 Vollgraf, U., Wegner, M., and Richter-Landsberg, C (1999) Activation of AP-1 and nuclear factor-kappaB transcription factors is involved in hydrogen peroxide-induced apoptotic cell death of oligodendrocytes J Neurochem 73, 2501-2509 Wade, C., and Dorsa, D (2003) Estrogen activation of cyclic adenosine 5'-monophosphate response element-mediated transcription requires the extracellularly regulated kinase/mitogen-activated protein kinase pathway Endocrinology 144, 832-838 Wakabayashi, S., Bertrand, B., Ikeda, T., Pouysségur, J., and Shigekawa, M (1994) Mutation of calmodulin-binding site renders the Na+/H+ exchanger (NHE1) highly H(+)-sensitive and Ca2+ regulation-defective J Biol Chem 269, 13710-13715 Walter, P., Green, S., Greene, G., Krust, A., Bornert, J M., Jeltsch, J M., Staub, A., Jensen, E., Scrace, G., and Waterfield, M (1985) Cloning of the human estrogen receptor cDNA Proc Natl Acad Sci U S A 82, 7889-7893 Wang, C., Fu, M., Angeletti, R., Siconolfi-Baez, L., Reutens, A., Albanese, C., Lisanti, M., Katzenellenbogen, B., Kato, S., and Hopp, T., et al (2001) Direct acetylation of the estrogen receptor alpha hinge region by p300 regulates transactivation and hormone sensitivity J Biol Chem 276, 18375-18383 Wang, G., Jiang, B., and Semenza, G (1995) Effect of altered redox states on expression and DNA-binding activity of hypoxia-inducible factor Biochem Biophys Res Commun 212, 550-556 Wang, H., Singh, D., and Fliegel, L (1997) The Na+/H+ antiporter potentiates growth and retinoic acid-induced differentiation of P19 embryonal carcinoma cells J Biol Chem 272, 26545-26549 Wang, J., and Yi, J (2008) Cancer cell killing via ROS: to increase or decrease, that is the question Cancer Biol Ther 7, 1875-1884 Wang, T., Xu, J., Yu, X., Yang, R., and Han, Z (2006) Peroxisome proliferator-activated receptor gamma in malignant diseases Crit Rev Oncol Hematol 58, 1-14 Wang, Y., and Casero, R (2006) Mammalian polyamine catabolism: a therapeutic target, a pathological problem, or both? J Biochem 139, 17-25 Wang, Y., Zong, H., Chi, Y., Hong, Y., Yang, Y., Zou, W., Yun, X., and Gu, J (2009) Repression of estrogen receptor alpha by CDK11p58 through promoting its ubiquitinproteasome degradation J Biochem 145, 331-343 Wang, Z., Orlowski, J., and Shull, G (1993) Primary structure and functional expression of a novel gastrointestinal isoform of the rat Na/H exchanger J Biol Chem 268, 11925-11928 Ward, N., Pierce, D., Chung, S., Gravitt, K., and O'Brian, C (1998) Irreversible inactivation of protein kinase C by glutathione J Biol Chem 273, 12558-12566 Ward, N., Stewart, J., Ioannides, C., and O'Brian, C (2000) Oxidant-induced Sglutathiolation inactivates protein kinase C-alpha (PKC-alpha): a potential mechanism of PKC isozyme regulation Biochemistry 39, 10319-10329 Washburn, T., Hocutt, A., Brautigan, D., and Korach, K (1991) Uterine estrogen receptor in vivo: phosphorylation of nuclear specific forms on serine residues Mol Endocrinol 5, 235242 232 Watanabe, T., Inoue, S., Ogawa, S., Ishii, Y., Hiroi, H., Ikeda, K., Orimo, A., and Muramatsu, M (1997) Agonistic effect of tamoxifen is dependent on cell type, ERE-promoter context, and estrogen receptor subtype: functional difference between estrogen receptors alpha and beta Biochem Biophys Res Commun 236, 140-145 Webb, P., Lopez, G., Uht, R., and Kushner, P (1995) Tamoxifen activation of the estrogen receptor/AP-1 pathway: potential origin for the cell-specific estrogen-like effects of antiestrogens Mol Endocrinol 9, 443-456 Webb, P., Nguyen, P., Valentine, C., Lopez, G., Kwok, G., McInerney, E., Katzenellenbogen, B., Enmark, E., Gustafsson, J., and Nilsson, S., et al (1999) The estrogen receptor enhances AP-1 activity by two distinct mechanisms with different requirements for receptor transactivation functions Mol Endocrinol 13, 1672-1685 Webster, R., Brockman, D., and Myatt, L (2006) Nitration of p38 MAPK in the placenta: association of nitration with reduced catalytic activity of p38 MAPK in pre-eclampsia Mol Hum Reprod 12, 677-685 Weigel, M., and Dowsett, M (2010) Current and emerging biomarkers in breast cancer: prognosis and prediction Endocr Relat Cancer 17, R245-R262 Weitsman, G., Weebadda, W., Ung, K., and Murphy, L (2009) Reactive oxygen species induce phosphorylation of serine 118 and 167 on estrogen receptor alpha Breast Cancer Res Treat 118, 269-279 Whittal, R., Benz, C., Scott, G., Semyonov, J., Burlingame, A., and Baldwin, M (2000) Preferential oxidation of zinc finger in estrogen receptor DNA-binding domain prevents dimerization and, hence, DNA binding Biochemistry 39, 8406-8417 Wierstra, I (2008) Sp1: emerging roles beyond constitutive activation of TATA-less housekeeping genes Biochem Biophys Res Commun 372, 1-13 Wijayaratne, A., and McDonnell, D (2001) The human estrogen receptor-alpha is a ubiquitinated protein whose stability is affected differentially by agonists, antagonists, and selective estrogen receptor modulators J Biol Chem 276, 35684-35692 Woo, H., Yim, S., Shin, D., Kang, D., Yu, D., and Rhee, S (2010) Inactivation of peroxiredoxin I by phosphorylation allows localized H(2)O(2) accumulation for cell signaling Cell 140, 517-528 Woo, S., Park, I., Park, M., An, S., Lee, H., Jin, H., Park, S., Cho, H., Lee, S., and Gwak, H., et al (2004) Arsenic trioxide sensitizes CD95/Fas-induced apoptosis through ROS-mediated upregulation of CD95/Fas by NF-kappaB activation Int J Cancer 112, 596-606 Woodfield, G., Horan, A., Chen, Y., and Weigel, R (2007) TFAP2C controls hormone response in breast cancer cells through multiple pathways of estrogen signaling Cancer Res 67, 8439-8443 Wyckoff, M., Chambliss, K., Mineo, C., Yuhanna, I., Mendelsohn, M., Mumby, S., and Shaul, P (2001) Plasma membrane estrogen receptors are coupled to endothelial nitric-oxide synthase through Galpha(i) J Biol Chem 276, 27071-27076 Yan, W., Nehrke, K., Choi, J., and Barber, D (2001) The Nck-interacting kinase (NIK) phosphorylates the Na+-H+ exchanger NHE1 and regulates NHE1 activation by plateletderived growth factor J Biol Chem 276, 31349-31356 233 Yang, W., Dyck, J., Wang, H., and Fliegel, L (1996) Regulation of NHE-1 promoter in mammalian myocardium Am J Physiol 270, H259-H266 Yang, W., Wang, H., and Fliegel, L (1996) Regulation of Na+/H+ exchanger gene expression Role of a novel poly(dA.dT) element in regulation of the NHE1 promoter J Biol Chem 271, 20444-20449 Yang, X., Ferguson, A., Nass, S., Phillips, D., Butash, K., Wang, S., Herman, J., and Davidson, N (2000) Transcriptional activation of estrogen receptor alpha in human breast cancer cells by histone deacetylase inhibition Cancer Res 60, 6890-6894 Yudt, M., Vorojeikina, D., Zhong, L., Skafar, D., Sasson, S., Gasiewicz, T., and Notides, A (1999) Function of estrogen receptor tyrosine 537 in hormone binding, DNA binding, and transactivation Biochemistry 38, 14146-14156 Yue, W., Santen, R., Wang, J., Li, Y., Verderame, M., Bocchinfuso, W., Korach, K., Devanesan, P., Todorovic, R., and Rogan, E., et al (2003) Genotoxic metabolites of estradiol in breast: potential mechanism of estradiol induced carcinogenesis J Steroid Biochem Mol Biol 86, 477-486 Zhou, Y., Hileman, E., Plunkett, W., Keating, M., and Huang, P (2003) Free radical stress in chronic lymphocytic leukemia cells and its role in cellular sensitivity to ROS-generating anticancer agents Blood 101, 4098-4104 Zou, A., Marschke, K., Arnold, K., Berger, E., Fitzgerald, P., Mais, D., and Allegretto, E (1999) Estrogen receptor beta activates the human retinoic acid receptor alpha-1 promoter in response to tamoxifen and other estrogen receptor antagonists, but not in response to estrogen Mol Endocrinol 13, 418-430 Zouki, C., Zhang, S., Chan, J., and Filep, J (2001) Peroxynitrite induces integrin-dependent adhesion of human neutrophils to endothelial cells via activation of the Raf-1/MEK/Erk pathway FASEB J 15, 25-27 de, L., and Villegas, I (2007) Resveratrol as an antioxidant and pro-oxidant agent: mechanisms and clinical implications Biochem Soc Trans 35, 1156-1160 deConinck, E C., McPherson, L A., and Weigel, R J (1995) Transcriptional regulation of estrogen receptor in breast carcinomas Mol Cell Biol 15, 2191-2196 deGraffenried, L., Hilsenbeck, S., and Fuqua, S (2002) Sp1 is essential for estrogen receptor alpha gene transcription J Steroid Biochem Mol Biol 82, 7-18 deGraffenried, L., Hopp, T., Valente, A., Clark, R., and Fuqua, S (2004) Regulation of the estrogen receptor alpha minimal promoter by Sp1, USF-1 and ERalpha Breast Cancer Res Treat 85, 111-120 234 ... REGULATION OF NHE1…………… ? ?11 0 3B .1 DOWN -REGULATION OF NHE1 BY H2O2 11 0 3B .1. 1 Identification of crucial region in the promoter of NHE1 11 0 3B .1. 2 H2O2 down-regulates 0 .15 kb proximal NHE1 promoter... signaling 21 1.3.4 Regulation of estrogen receptors 23 1. 3.5 Estrogen and estrogen receptors in human breast cancer 27 v 1. 4 THE SODIUM HYDROGEN EXCHANGER 29 1. 4 .1 pH regulation. .. turnover 16 1. 3 ESTROGEN RECEPTOR 17 1. 3 .1 Estrogen and estrogen receptors: Introduction 17 1. 3.2 Estrogen receptor and genomic signaling 20 1. 3.3 Membrane and cytoplasmic