The depletion of p38alpha kinase upregulates NADPH oxidase 2/NOX2/gp91 expression and the production of superoxide in mouse embryonic stem cells

• Published in: Archives of biochemistry and biophysics 2019-08, Vol.671, p.18-26
• Main Authors: Binó, Lucia, Veselá, Iva, Papežíková, Iva, Procházková, Jiřina, Vašíček, Ondřej, Štefková, Kateřina, Kučera, Jan, Hanáčková, Markéta, Kubala, Lukáš, Pacherník, Jiří
• Format: Article
• Language: English
• Subjects: Animals; Cell Differentiation - physiology; Cells, Cultured; Embryonic stem cell; Gene Knockdown Techniques; Gene Knockout Techniques; Membrane Potential, Mitochondrial - physiology; Mice; Mitochondria - metabolism; Mitogen-Activated Protein Kinase 14 - genetics; Mitogen-Activated Protein Kinase 14 - metabolism; Mouse Embryonic Stem Cells - metabolism; NADPH oxidase; NADPH Oxidase 2 - genetics; NADPH Oxidase 2 - metabolism; p38 kinase; Reactive oxygen species; Superoxides - metabolism
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1016/j.abb.2019.06.001

P38alpha kinase plays an important role in the regulation of both cell stress response and cell fate. In this study, we report that p38alpha kinase-deficient embryonic stem cells exhibit a higher production of reactive oxygen species (ROS) in contrast to their wild-type counterpart. Analysis of the expressions of NADPH oxidases (NOXs) and dual oxidases, crucial enzymes involved in intracellular ROS formation, shows NOX2/gp91phox is over-expressed in p38alpha deficient cells. The particular increase in superoxide formation was confirmed by the specific detection of hydroethidine derivate 2-hydroxyethidium. ROS formation decreased when the level of NOX2 was silenced by siRNA in p38alpha deficient cells. These data suggest the importance of p38alpha kinase in the regulation of ROS metabolism in embryonic stem cells and the significance of the observed phenomena of cancer cell-like phenotypes, which is discussed. •Production of superoxide is increased in p38a-deficient embryonic stem (ES) cells.•P38a-deficient ES cells express high levels of NOX2.•Downregulation of NOX2 by siRNA leads to decreased superoxide levels in p38a-deficient ES cells.•P38 kinase regulates NOX2 expression and superoxide production in ES cells.