Generation of Reactive Oxygen Species in a Human Keratinocyte Cell Line: Role of Calcium

Goldman, Rachel; Moshonov, Sandra; Zor, U.

doi:10.1006/abbi.1997.0478

Cited by 71 publications

(44 citation statements)

References 41 publications

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“…We also demonstrated that Duox1 activation in A431 and HaCaT cells is a consequence of an increase in intracellular calcium, elicited by EGF. Our data provide the missing link between agonist-evoked calcium signals and ROS production, which were previously described to be connected in epidermal cell lines (9). EGF-induced H 2 O 2 production by A431 cells was originally described by Bae et al (8) and was subsequently confirmed by several other groups (22), (23).…”

Section: Discussionsupporting

confidence: 69%

“…There are multiple stimuli that can trigger the formation of H 2 O 2 in mammalian cells and growth factor-induced H 2 O 2 production is thought to be a particularly important signaling event (6), (7). EGF-induced H 2 O 2 production was first described in A431 cells and has been subsequently demonstrated in other cell types as well (8), (9). The ROS response elicited by EGF is thought to amplify tyrosine-kinase signaling through the inhibition of protein tyrosine phosphatases that normally antagonize this signaling route (10), (7).…”

Section: Introductionmentioning

confidence: 99%

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Epidermal growth factor-induced hydrogen peroxide production is mediated by dual oxidase 1

Sirokmány

Pató

Zana

et al. 2016

Free Radical Biology and Medicine

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Stimulation of mammalian cells by epidermal growth factor (EGF) elicits complex signaling events, including an increase in hydrogen peroxide (H 2 O 2 ) production. Understanding the significance of this response is limited by the fact that the source of EGF-induced H 2 O 2 production is unknown. Here we show that EGF-induced H 2 O 2 production in epidermal cell lines is dependent on the agonist-induced calcium signal. We analyzed the expression of NADPH oxidase isoforms and found both A431 and HaCaT cells to express the calcium-sensitive NADPH oxidase, Dual oxidase 1 (Duox1) and its protein partner Duox activator 1 (DuoxA1).Inhibition of Duox1 expression by small interfering RNAs eliminated EGF-induced H 2 O 2 production in both cell lines. We also demonstrate that H 2 O 2 production by Duox1 leads to the oxidation of thioredoxin-1 and the cytosolic peroxiredoxins. Our observations provide evidence for a new signaling paradigm in which changes of intracellular calcium concentration are transformed into redox signals through the calcium-dependent activation of Duox1.

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Section: Discussionsupporting

confidence: 69%

Section: Introductionmentioning

confidence: 99%

Epidermal growth factor-induced hydrogen peroxide production is mediated by dual oxidase 1

Sirokmány

Pató

Zana

et al. 2016

Free Radical Biology and Medicine

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“…Figure 1a shows that the intensity of the chemiluminescence signal increased as the concentration of LPA increased and saturated at 5 µM LPA. Although the chemiluminescence signal varied between experiments as observed previously [5], it was quite consistent within the same experiment. In a standard assay using 1i10' cells, 5 µM LPA typically produced a maximal chemiluminescence signal between 4i10' rlu\s and 8i10' rlu\s.…”

Section: Lpa Stimulates H 2 O 2 Release In Hacat Cellssupporting

confidence: 86%

“…Cells were serum-starved for 24 h in serum-free DMEM prior to use for experiments. H # O # was determined by the luminol and HRP-dependent chemiluminescence assay [5]. Serum-starved cells were rinsed with PBS, trypsinized, suspended in Hanks balanced salt solution (HBSS), and were washed twice with HBSS.…”

Section: Cell Culture and H 2 O 2 Detectionmentioning

confidence: 99%

“…For example, plateletderived growth factor (PDGF) increases H # O # production in vascular smooth muscle cells [1], and basic fibroblast growth factor and tumour necrosis factor α increase ROS levels in chondrocytes [2]. Similarly, epidermal growth factor (EGF) transiently induces H # O # production in human A431 epidermoid carcinoma cells, mouse NIH3T3 fibroblasts, and human HaCaT keratinocytes [3][4][5]. Increased amounts of H # O # have also been detected in HeLa cells treated with lysophosphatidic acid (LPA) [6].…”

Section: Introductionmentioning

confidence: 99%

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Involvement of lipoxygenase in lysophosphatidic acid-stimulated hydrogen peroxide release in human HaCaT keratinocytes

Sekharam

Cunnick

2000

Biochemical Journal

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Although it is now recognized that low levels of reactive oxygen species (ROS) are required for the mitogenic response, mitogen-induced signalling pathways that regulate ROS generation in non-phagocytic cells remain largely uncharacterized. Using a real-time assay for measuring hydrogen peroxide (H2O2) formation, we analysed H2O2 release in human HaCaT keratinocytes in response to lysophosphatidic acid (LPA), a mitogen for keratinocytes. LPA rapidly increased H2O2 release in HaCaT cells. Unlike LPA-induced mitogen-activated protein (MAP) kinase activation, LPA-stimulated H2O2 release was independent of the tyrosine kinase activity of the epidermal growth factor (EGF) receptor. Calcium chelators, phospholipase A2 inhibitors, and lipoxygenase inhibitors effectively blocked LPA-stimulated H2O2 release, whereas cyclooxygenase inhibitors were without effect. Addition of 5-lipoxygenase products 5-hydroperoxyeicosatetraenoic acid (5-HPETE) and leukotriene B4, but not 5-hydroxyeicosatetraenoic acid (5-HETE) and leukotriene C4, restored LPA-stimulated H2O2 release in cells treated with the lipoxygenase inhibitors nordihydroguaiaretic acid and Zileuton. These results suggest that the lipoxygenase products 5-HPETE and leukotriene B4 are required for LPA-stimulated H2O2 release in HaCaT cells.

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Biostimulative effects of low-energy lasers and their implications for medicine

Lubart

Breitbart

2000

Drug Dev. Res.

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Generation of Reactive Oxygen Species in a Human Keratinocyte Cell Line: Role of Calcium

Cited by 71 publications

References 41 publications

Epidermal growth factor-induced hydrogen peroxide production is mediated by dual oxidase 1

Epidermal growth factor-induced hydrogen peroxide production is mediated by dual oxidase 1

Involvement of lipoxygenase in lysophosphatidic acid-stimulated hydrogen peroxide release in human HaCaT keratinocytes

Biostimulative effects of low-energy lasers and their implications for medicine

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