Development of High-density DNA Microarray Membrane for Profiling Smoke- and Hydrogen Peroxide-induced Genes in a Human Bronchial Epithelial Cell Line

Yoneda, Ken Y.; Peck, Konan; Chang, Melinda Y.; Chmiel, K.; Sher, Yuh‐Pyng; Chen, Jeremy; Yang, Pan‐Chyr; Chen, Yin; Wu, Reen

doi:10.1164/ajrccm.164.supplement_2.2106062

Cited by 42 publications

(30 citation statements)

References 29 publications

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“…MKP-1 is constitutively expressed at a very low level and underlies a tight and rapid transcriptional induction by different stimuli. 7 Little is known about the signaling events leading to an MKP-1 induction besides reports describing that reactive oxygen species (ROS) 8,9 and different kinases (MAPK and protein kinase C [PKC]) 10 -13 might be involved. For several decades, ROS were considered toxic byproducts of metabolic processes being able to damage cellular components.…”

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confidence: 99%

Atrial Natriuretic Peptide Induces Mitogen-Activated Protein Kinase Phosphatase-1 in Human Endothelial Cells via Rac1 and NAD(P)H Oxidase/Nox2-Activation

Fürst

Brueckl

Kuebler

et al. 2005

Circulation Research

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Abstract-The cardiovascular hormone atrial natriuretic peptide (ANP) exerts anti-inflammatory effects on tumor necrosis factor-␣-activated endothelial cells by inducing mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1). The underlying mechanisms are as yet unknown. We aimed to elucidate the signaling pathways leading to an induction of MKP-1 by ANP in primary human endothelial cells. By using antioxidants, generation of reactive oxygen species (ROS) was shown to be crucially involved in MKP-1 upregulation. ANP was found to increase ROS formation in cultured cells as well as in the endothelium of intact rat lung vessels. We applied NAD(P)H oxidase (Nox) inhibitors (apocynin and gp91ds-tat) and revealed this enzyme complex to be crucial for superoxide generation and MKP-1 expression. Moreover, by performing Nox2/4 antisense experiments, we identified Nox2 as the critically involved Nox homologue. Pull-down assays and confocal microscopy showed that ANP activates the small Rho-GTPase Rac1. Transfection of a dominant-negative (RacN17) and constitutively active Rac1 mutant (RacV12) indicated that ANP-induced superoxide generation and MKP-1 expression are mediated via Rac1 activation. ANP-evoked production of superoxide was found to activate c-Jun N-terminal kinase (JNK). Using specific inhibitors, we linked ANP-induced JNK activation to MKP-1 expression and excluded an involvement of protein kinase C, extracellular signal-regulated kinase, and p38 MAPK. MKP-1 induction was shown to depend on activation of the transcription factor activator protein-1 (AP-1) by using electrophoretic mobility shift assay and AP-1 decoys. In summary, our work provides insights into the mechanisms by which ANP induces MKP-1 and shows that ANP is a novel endogenous activator of endothelial Rac1 and Nox/Nox2. T he cardiovascular hormone atrial natriuretic peptide (ANP) plays an important and well-investigated role in the cardiovascular system by participation in blood pressure regulation. 1 ANP mainly acts through binding to the guanylyl cyclase-coupled natriuretic peptide receptor-A (NPR-A), leading to generation of the second messenger cGMP. 1 ANP also binds to NPR-C, which generally acts as a clearance receptor 1 but is also able to mediate an inhibition of adenylyl cyclase activity and activation of phospholipase C. 2 A growing number of studies highlight the profound anti-inflammatory and vasoprotective actions exerted by ANP. 3,4 In this context, we revealed recently that ANP is able to induce mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1) in cultured endothelial cells (ECs). 5 MKP-1 inhibits tumor necrosis factor-␣ (TNF-␣)-induced p38 MAPK activation, thus leading to a reduction of stress fiber formation, 5 endothelial permeability, 5 and expression of monocyte chemoattractant protein-1. 6 In summary, ANP protects TNF-␣-activated ECs against structural and functional changes by inducing MKP-1.MAPKs play a crucial role in a great variety of intracellular signaling cascades. Pathways of MAPK activation are qu...

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mentioning

confidence: 99%

Atrial Natriuretic Peptide Induces Mitogen-Activated Protein Kinase Phosphatase-1 in Human Endothelial Cells via Rac1 and NAD(P)H Oxidase/Nox2-Activation

Fürst

Brueckl

Kuebler

et al. 2005

Circulation Research

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“…A similar approach was also carried out in H 2 O 2 -treated cells. Twenty-two genes were viewed as inducible genes (27), and 14 of these genes were further confirmed by Northern blot hybridization; all of them were commonly elevated in smoke-and H 2 O 2 -exposed cells. A time course Northern blot hybridization study was carried out to elucidate the time course induction by smoke (Figure 3).…”

Section: Resultsmentioning

confidence: 90%

“…From this study, three phases of gene induction can be assessed (27). The first one is an immediate phase, in which inducible genes could be seen within 1 h after smoke exposure.…”

Section: Discussionmentioning

confidence: 99%

Application of High-Density DNA Microarray to Study Smoke- and Hydrogen Peroxide–Induced Injury and Repair in Human Bronchial Epithelial Cells

Yoneda

Chang

Chmiel

et al. 2003

Journal of the American Society of Nephrology

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ABSTRACT. Recent advances in high-density DNA microarray technique allow the possibility to analyze thousands of genes simultaneously for their differential gene expression patterns in various biologic processes. Through clustering analysis and pattern recognition, the significance of these differentially expressed genes can be recognized and correlated with the biologic events that may take place inside the cell and tissue. High-density DNA microarray nylon membranes were used to explore gene expression and regulation associated with smoke- and hydrogen peroxide–induced injury and repair in differentiated human bronchial epithelial cells in vitro. At least three phases of change in gene expression could be recognized. The first phase seems to be an immediate event in response to oxidant injury. This phase includes the induction of bcl-2 and mdm2 genes that are involved in the regulation of apoptosis, and the mitogen-activated protein kinase phosphatase 1 that functions as a regulator for various mitogen-activated protein kinase activities. The second phase, usually 5 h later, includes the induction of various stress proteins and ubiquitin, which are important in providing the chaperone mechanism and the turnover of damaged macromolecules. The third phase, which is 5 to 10 h later, includes the induction of genes that seem to be involved in reducing oxidative stress by metabolizing the cellular level of reactive oxygen species. In this phase, enzymes associated with tissue and cell remodeling are also elevated. These results demonstrated a complex gene expression array by bronchial epithelial cells in response to a single insult of oxidants that are relevant to environmental pollutants. E-mail: rwu@ucdavis.edu

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“…In lung epithelial cells, oxidative damage to DNA is caused not only by air pollutants, but also by neutrophil products [19]. Using high-density DNA microarrays, lung epithelial cells exposed to oxidative injury showed early induction of bcl-2 and mdm-2, genes which are known to be are involved in the regulation of apoptosis [20]. Both DNA and mitochondrial events combine to regulate cell cycle progression and apoptosis after treatment with H 2 O 2 .…”

Section: Discussionmentioning

confidence: 99%

Dissociation of DNA damage and mitochondrial injury caused by hydrogen peroxide in SV-40 transformed lung epithelial cells

et al. 2002

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BackgroundSince lung epithelial cells are constantly being exposed to reactive oxygen intermediates (ROIs), the alveolar surface is a major site of oxidative stress, and each cell type may respond differently to oxidative stress. We compared the extent of oxidative DNA damage with that of mitochondrial injury in lung epithelial cells at the single cell level.ResultDNA damage and mitochondrial injury were measured after oxidative stress in the SV-40 transformed lung epithelial cell line challenged with hydrogen peroxide (H2O2). Single cell analysis of DNA damage was determined by assessing the number of 8-oxo-2-deoxyguanosine (8-oxo-dG) positive cells, a marker of DNA modification, and the length of a comet tail. Mitochondrial membrane potential, ΔΨm, was determined using JC-1. A 1 h pulse of H2O2 induced small amounts of apoptosis (3%). 8-oxo-dG-positive cells and the length of the comet tail increased within 1 h of exposure to H2O2. The number of cells with reduced ΔΨm increased after the addition of H2O2 in a concentration-dependent manner. In spite of a continual loss of ΔΨm, DNA fragmentation was reduced 2 h after exposure to H2O2.ConclusionThe data suggest that SV-40 transformed lung epithelial cells are resistant to oxidative stress, showing that DNA damage can be dissociated from mitochondrial injury.

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Development of High-density DNA Microarray Membrane for Profiling Smoke- and Hydrogen Peroxide-induced Genes in a Human Bronchial Epithelial Cell Line

Cited by 42 publications

References 29 publications

Atrial Natriuretic Peptide Induces Mitogen-Activated Protein Kinase Phosphatase-1 in Human Endothelial Cells via Rac1 and NAD(P)H Oxidase/Nox2-Activation

Atrial Natriuretic Peptide Induces Mitogen-Activated Protein Kinase Phosphatase-1 in Human Endothelial Cells via Rac1 and NAD(P)H Oxidase/Nox2-Activation

Application of High-Density DNA Microarray to Study Smoke- and Hydrogen Peroxide–Induced Injury and Repair in Human Bronchial Epithelial Cells

Dissociation of DNA damage and mitochondrial injury caused by hydrogen peroxide in SV-40 transformed lung epithelial cells

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