Journalartikel

Jahr der Veröffentlichung: 2016

Seiten: 159-174

Zeitschrift: Cell and Tissue Research

Bandnummer: 364

Heftnummer: 1

ISSN: 0302-766X

eISSN: 1432-0878

DOI Link: https://doi.org/10.1007/s00441-015-2303-8

Verlag: Springer

Abstract: 
The impact of reactive oxygen species and phosphoinositide 3-kinase (PI3K) in differentiating embryonic stem (ES) cells is largely unknown. Here, we show that the silencing of the PI3K catalytic subunit p110 alpha and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 1 (NOX1) by short hairpin RNA or pharmacological inhibition of NOX and ras-related C3 botulinum toxin substrate 1 (Rac1) abolishes superoxide production by vascular endothelial growth factor (VEGF) in mouse ES cells and in ES-cell-derived fetal liver kinase-1(+) (Flk-1(+)) vascular progenitor cells, whereas the mitochondrial complex I inhibitor rotenone does not have an effect. Silencing p110 alpha or inhibiting Rac1 arrests vasculogenesis at initial stages in embryoid bodies, even under VEGF treatment, as indicated by platelet endothelial cell adhesion molecule-1 (PECAM-1)-positive areas and branching points. In the absence of p110 alpha, tube-like structure formation on matrigel and cell migration of Flk-1(+) cells in scratch migration assays are totally impaired. Silencing NOX1 causes a reduction in PECAM-1-positive areas, branching points, cell migration and tube length upon VEGF treatment, despite the expression of vascular differentiation markers. Interestingly, silencing p110 alpha but not NOX1 inhibits the activation of Rac1, Ras homologue gene family member A (RhoA) and Akt leading to the abrogation of VEGF-induced lamellipodia structure formation. Thus, our data demonstrate that the PI3K p110 alpha-Akt/Rac1 and NOX1 signalling pathways play a pivotal role in VEGF-induced vascular differentiation and cell migration. Rac1, RhoA and Akt phosphorylation occur downstream of PI3K and upstream of NOX1 underscoring a role of PI3K p110 alpha in the regulation of cell polarity and migration.

Harvard-Zitierstil: Bekhite, M., Mueller, V., Troeger, S., Mueller, J., Figulla, H., Sauer, H., et al. (2016) Involvement of phosphoinositide 3-kinase class IA (PI3K 110α) and NADPH oxidase 1 (NOX1) in regulation of vascular differentiation induced by vascular endothelial growth factor (VEGF) in mouse embryonic stem cells, Cell and Tissue Research, 364(1), pp. 159-174. https://doi.org/10.1007/s00441-015-2303-8

APA-Zitierstil: Bekhite, M., Mueller, V., Troeger, S., Mueller, J., Figulla, H., Sauer, H., & Wartenberg, M. (2016). Involvement of phosphoinositide 3-kinase class IA (PI3K 110α) and NADPH oxidase 1 (NOX1) in regulation of vascular differentiation induced by vascular endothelial growth factor (VEGF) in mouse embryonic stem cells. Cell and Tissue Research. 364(1), 159-174. https://doi.org/10.1007/s00441-015-2303-8