Hypoxia activates signal transducers and activators of transcription 5 (STAT5) and increases its binding activity to the GAS element in mammary epithelial cells

Joung, Youn-Hee; Park, Jong‐Hwan; Park, Taekyu; Lee, Chang‐Soo; Kim, Oun Hyun; Ye, Sang‐Kyu; Yang, U. M.; Lee, Kwang Jeon; Yang, Young Mok

doi:10.1038/emm.2003.46

Cited by 34 publications

(37 citation statements)

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“…The expression vectors for mouse STAT5a and STAT5b (pMX/STAT5a and pMX/STAT5b; kindly provided by Dr. Koichi Ikuta, Kyoto University, Japan) were constructed as previously described (Joung et al, 2003). A series of 5' promoter reporter constructions derived from the human cyclin D1 genomic clone were previously described (Albanese et al, 1995).…”

Section: Expression Vectors and Construction Of Plasmid Vectormentioning

confidence: 99%

“…For reporter gene assays, COS-7 cells were transiently co-transfected with the cyclin D1 (GAS1) construct (Albanese et al, 1995) and the STAT5a or STAT5b expression vector (Joung et al, 2003). Transfected cells were washed twice with ice-cold PBS, and 150 l of lysis buffer was added to the wells.…”

Section: Luciferase Assaysmentioning

confidence: 99%

“…The WCE (20 l per each lane) or immunoprecipitated proteins were subjected to SDS-PAGE and electrophoretically transferred onto a nitrocellulose membrane. Western blots were performed as described previously (Joung et al, 2003). After blocking the residual binding sites on the filter, immunoblotting was performed with an appropriate antibody.…”

Section: Immunoprecipitation and Western Blottingmentioning

confidence: 99%

“…The double-stranded DNA probe was end-labeled using T4 polynucleotide kinases and [ -32 P]ATP and used in all EMSAs. EMSAs were performed as described previously (Joung et al, 2003). For supershift assays, 1 g of antibody was incubated with nuclear extracts for 20 min on ice before the addition of radiolabeled probe.…”

Section: Oligonucleotide Labeling and Electrophoretic Mobility Shift mentioning

confidence: 99%

“…For instance, STAT5b is selectively important for growth hormone signaling (Udy et al, 1997), whereas STAT5a is particularly critical for prolactin-induced mammary gland differentiation (Liu et al, 1997;Yamashita et al, 2001). We recently demonstrated that desferrioxamine (DFO) or hypoxia activates STAT5 and enhances its binding to the IFN-activated sequence (GAS) sequence in the -casein gene promoter in mammary epithelial cells (Joung et al, 2003). DFO, that mimics hypoxia and a similar oxygen sensor in the hypoxia regulation, induces a hypoxic condition (Tazuke et al, 1998;Park et al, 2000;Park et al, 2001a).…”

Section: Introductionmentioning

confidence: 99%

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Hypoxia activates the cyclin D1 promoter via the Jak2/STAT5b pathway in breast cancer cells

Joung

Lim²,

Lee³

et al. 2005

Exp Mol Med

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Hypoxia, a common consequence of solid tumor growth in breast cancer or other cancers, serves to propagate a cascade of molecular pathways which include angiogenesis, glycolysis, and various cellcycle control proteins. As we have shown previously, hypoxia activates STAT5 (signal transducer and activator of transcription 5) and increases its binding activity to the GAS element in mammary epithelial cells. In this study we attempted to elucidate the mechanism by which cyclin D1 is regulated by the STAT5 protein under hypoxic conditions. Our data demonstrate that hypoxia (2% O 2 ) or desferrioxamine (DFO) induces tyrosine and serine phosphorylation of STAT5 in human breast cancer cells (MCF-7) and mammary epithelial cells (HC11). Imunoprecipitation and subsequent Western analysis showed that Jak2 leads to the tyrosine phosphorylation and activation of STAT5a or STAT5b under hypoxic conditions. Using a transfected COS-7 cell model system, we demonstrate that the activity of a cyclin D1 promoter-luciferase construct increased under hypoxic conditions or DFO treatment. The activity of the STAT5b/cyclin D1 promoter increased significantly by 12 h of hypoxia, whereas the activity of the STAT5a/cyclin D1 promoter was unaffected under hypoxic conditions. These increases in promoter activity are predominantly mediated by the Jak2/ STAT5b signaling pathway. We have shown by EMSA that hypoxia induces STAT5 to bind to the cyclin D1 promoter (GAS-1) in MCF-7 and HC11 cells. These data suggest that STAT5b may mediate the transcriptional activation of cyclin D1 after hypoxic stimulation.

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Section: Expression Vectors and Construction Of Plasmid Vectormentioning

confidence: 99%

Section: Luciferase Assaysmentioning

confidence: 99%

Section: Immunoprecipitation and Western Blottingmentioning

confidence: 99%

Section: Oligonucleotide Labeling and Electrophoretic Mobility Shift mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Hypoxia activates the cyclin D1 promoter via the Jak2/STAT5b pathway in breast cancer cells

Joung

Lim²,

Lee³

et al. 2005

Exp Mol Med

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Pathophysiological hypoxia affects the redox state and IL‐2 signalling of human CD4⁺ T cells and concomitantly impairs survival and proliferation

et al. 2013

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Inflamed areas are characterized by infiltration of immune cells, local hypoxia and alterations of cellular redox states. We investigated the impact of hypoxia on survival, proliferation, cytokine secretion, intracellular energy and redox state of human CD4+ T cells.We found that pathophysiological hypoxia (<2% O 2 ) significantly decreased CD4 + T-cell survival after mitogenic stimulation. This effect was not due to an increased caspase-3/7-mediated apoptosis or adenosine-5 -triphosphate (ATP) consumption/depletion. However, the ability of stimulated T cells to proliferate was reduced under hypoxic conditions, despite increased expression of CD25. Pathophysiological hypoxia was also found to modify intracellular ROS (iROS) levels in stimulated T cells over time as compared with levels found in normoxia. Physiological hypoxia (5% O 2 ) did not decrease CD4 + T-cell survival and proliferation or modify iROS levels as compared with normoxia. We conclude that pathophysiological hypoxia affects T-cell proliferation and viability via disturbed IL-2R signalling downstream of STAT5a phosphorylation, but not as a result of impaired cellular energy homeostasis. We suggest iROS links early events in T-cell stimulation to the inhibition of the lymphoproliferative response under pathophysiological hypoxic conditions. The level of iROS may therefore act as a mediator of immune functions leading to down-regulation of long-term T-cell activity in inflamed tissues.Keywords: Bioenergetics r CD4 + T cells r Hypoxia r ROS IntroductionLocal pathophysiological hypoxia is associated with the pathogenesis of arthritis [1], sepsis [2], ischemic areas after embolic ischemic stroke [3], fracture haematoma [4,5] and in fast growingCorrespondence: Dr. Timo Gaber e-mail: gaber@drfz.de tumour tissue [6]. In these areas of compromised oxygen supply, immune cells are able to be functionally active [7]. Pathophysiological hypoxia (<2% O 2 ) is known to enhance the proinflammatory potential of myeloid cells such as neutrophils and macrophages [8], but lack of oxygen also acts in a tissue * These authors contributed equally to this work.C 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.eji-journal.eu Eur. J. Immunol. 2013. 43: 1588-1597 Immunomodulation 1589 protective manner and suppresses typical T-cell effector functions, such as production of cytokines and T-cell proliferation [9,10]. Early observations reported that hypoxia stimulates Con A-induced PBMC proliferation and increases IL-2 expression [11]. These data were partially confirmed by Naldini et al. [12,13] using PHA-stimulated PBMCs, although these authors reported a delay of cell cycle progression. Moreover, and observed an increase in PHA/IL-2-driven or TCR/IL-2-driven T-cell proliferation under hypoxia, an inhibition of apoptosis and an increased expression of CD25 per cells. These data on T-cell proliferation were confirmed by Bohnenkamp et al. [17]. However, Loeffler et al. [18] were the first to report that T cells exposed to hypoxia proliferate only weakly or not at all....

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Hypoxia-responsive transcription factors

Cummins

Taylor

2005

Pflugers Arch - Eur J Physiol

406

352

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Hypoxia is a common pathophysiological occurrence with a profound impact on the cellular transcriptome. The consequences of hypoxia-induced or hypoxia-repressed gene expression have important implications in disease processes as diverse as tumour development and chronic inflammation. While the hypoxia-inducible factor (HIF-1) plays a major role in controlling the ubiquitous transcriptional response to hypoxia, it is clear that a number of other transcription factors are also activated either directly or indirectly. In this review, we comprehensively discuss the transcription factors that have been reported to be hypoxia-responsive and the signalling mechanisms leading to their activation. Understanding such events will enhance our understanding of cellular oxygen sensing.

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Hypoxia activates signal transducers and activators of transcription 5 (STAT5) and increases its binding activity to the GAS element in mammary epithelial cells

Abstract: Abbreviations: DFO, desferrioxamine; EMSA, electrophoretic mobility shift assay; EPO, erythropoietin; GAS, interferon-γ activated site; HIF-1, hypoxia-inducible factor-1; PRL, prolactin; ROS, reactive oxygen species; STAT, signal transducers and activators of transcription

Cited by 34 publications

References 39 publications

Hypoxia activates the cyclin D1 promoter via the Jak2/STAT5b pathway in breast cancer cells

Hypoxia activates the cyclin D1 promoter via the Jak2/STAT5b pathway in breast cancer cells

Pathophysiological hypoxia affects the redox state and IL‐2 signalling of human CD4⁺ T cells and concomitantly impairs survival and proliferation

Hypoxia-responsive transcription factors

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Hypoxia activates signal transducers and activators of transcription 5 (STAT5) and increases its binding activity to the GAS element in mammary epithelial cells

Abstract: Abbreviations: DFO, desferrioxamine; EMSA, electrophoretic mobility shift assay; EPO, erythropoietin; GAS, interferon-γ activated site; HIF-1, hypoxia-inducible factor-1; PRL, prolactin; ROS, reactive oxygen species; STAT, signal transducers and activators of transcription

Cited by 34 publications

References 39 publications

Hypoxia activates the cyclin D1 promoter via the Jak2/STAT5b pathway in breast cancer cells

Hypoxia activates the cyclin D1 promoter via the Jak2/STAT5b pathway in breast cancer cells

Pathophysiological hypoxia affects the redox state and IL‐2 signalling of human CD4+ T cells and concomitantly impairs survival and proliferation

Hypoxia-responsive transcription factors

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Product

Resources

About

Pathophysiological hypoxia affects the redox state and IL‐2 signalling of human CD4⁺ T cells and concomitantly impairs survival and proliferation