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Cell Biochem Biophys. 2018 Sep;76(3):401-410. doi: 10.1007/s12013-018-0847-4. Epub 2018 Jun 29.

p47phox-Dependent Reactive Oxygen Species Stimulate Nuclear Translocation of the FoxO1 Transcription Factor During Metabolic Inhibition in Cardiomyoblasts.

Author information

1
Department of Cardiology, Amsterdam UMC, location AMC, Amsterdam, The Netherlands. e.terhorst@vumc.nl.
2
Netherlands Heart Institute, Utrecht, The Netherlands. e.terhorst@vumc.nl.
3
Amsterdam Cardiovascular Sciences, Amsterdam UMC, location VUmc, Amsterdam, The Netherlands. e.terhorst@vumc.nl.
4
Department of Pathology, Amsterdam UMC, location VUmc, Amsterdam, The Netherlands. e.terhorst@vumc.nl.
5
Amsterdam Cardiovascular Sciences, Amsterdam UMC, location VUmc, Amsterdam, The Netherlands.
6
Department of Pathology, Amsterdam UMC, location VUmc, Amsterdam, The Netherlands.
7
Department of Medical Biology L2-109, Amsterdam UMC, location AMC, Amsterdam, The Netherlands.
8
Department of Physiology, Amsterdam UMC, location VUmc, Amsterdam, The Netherlands.
9
Department of Cardiac Surgery, Amsterdam UMC, location VUmc, Amsterdam, The Netherlands.
10
Department of Cardiology, Amsterdam UMC, location VUmc, Amsterdam, The Netherlands.
11
Department of Cardiology, Amsterdam UMC, location AMC, Amsterdam, The Netherlands.

Abstract

Reactive oxygen species (ROS) control forkhead box O (FOXO) transcription factor activity by influencing their nuclear translocation. However, knowledge of the ROS cellular source(s) involved herein remains scarce. Recently, we have shown p47phox-dependent activation of ROS-producing NADPH oxidase (NOX) at the nuclear pore in H9c2 rat cardiomyoblasts in response to ischemia. This localizes NOX perfectly to affect protein nuclear translocation, including that of transcription factors. In the current study, involvement of p47phox-dependent production of ROS in the nuclear translocation of FOXO1 was analyzed in H9c2 cells following 4 h of metabolic inhibition (MI), which mimics the effects of ischemia. Nuclear translocation of FOXO1 was determined by quantitative digital-imaging fluorescence and western blot analysis. Subsequently, the effect of inhibiting p47phox-dependent ROS production by short hairpin RNA (shRNA) transfection on FOXO1 translocation was analyzed by digital-imaging microscopy. MI induced a significant translocation of FOXO1 into the nucleus. Transfection with p47phox-shRNA successfully knocked-down p47phox expression, reduced nuclear nitrotyrosine production, an indirect marker for ROS production, and inhibited the nuclear translocation of FOXO1 following MI. With these results, we show for the first time that nuclear import of FOXO1 induced by MI in H9c2 depends critically on p47phox-mediated ROS production.

KEYWORDS:

FOXO1; Ischemia; NADPH oxidase; Reactive oxygen species; p47phox

PMID:
29956081
PMCID:
PMC6097050
DOI:
10.1007/s12013-018-0847-4
[Indexed for MEDLINE]
Free PMC Article

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