title: Bidirectional cross-regulation between the endothelial nitric oxide synthase and β-catenin signalling pathways.
abstract: OBJECTIVE
  β-catenin has been shown to be regulated by inducible nitric oxide synthase (NOS) in endothelial cells. We investigated here whether β-catenin interacts with and regulates endothelial NOS (eNOS) and whether eNOS activation promotes β-catenin signalling.
  
  
  RESULTS
  We identified β-catenin as a novel eNOS binding protein in human umbilical vein endothelial cells (HUVECs) by mass spectroscopy and western blot analyses of β-catenin and eNOS immunoprecipitates. This was confirmed by in situ proximity ligation assay. eNOS activity, assessed by cGMP production and eNOS phosphorylation (Ser1177), was enhanced in β-catenin(-/-) mouse pulmonary endothelial cells (MPECs) relative to wild-type MPECs. eNOS activation (using adenosine, salbutamol, thrombin, or histamine), or application of an NO donor (spermine NONOate) or cGMP-analogue (8-bromo-cGMP) caused nuclear translocation of β-catenin in HUVEC as shown by western blotting of nuclear extracts. Exposure to spermine NONOate, 8-bromo-cGMP, or sildenafil (a phosphodiesterase type 5 inhibitor) also increased the expression of β-catenin-dependent transcripts, IL-8, and cyclin D1. Stimulation of wild-type MPECs with basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), spermine NONOate, 8-bromo-cGMP, or sildenafil increased tube length relative to controls in an angiogenesis assay. These responses were abrogated in β-catenin(-/-) MPECs, with the exception of that to bFGF which is NO-independent. In C57BL/6 mice, subcutaneous VEGF-supplemented Matrigel plugs containing β-catenin(-/-) MPECs exhibited reduced angiogenesis compared with plugs containing wild-type MPECs. Angiogenesis was not altered in bFGF-supplemented Matrigel.
  
  
  CONCLUSIONS
  These data reveal bidirectional cross-talk and regulation between the NO-cGMP and β-catenin signalling pathways.
authors: Cardiovascular Division, Department of Clinical Pharmacology, British Heart Foundation Centre of Research Excellence, King's College London, 3.07 Franklin-Wilkins Building, 150 Stamford Street, London, UK.
authors: Christina M
authors: CM
authors: Warboys
authors: Cardiovascular Division, Department of Clinical Pharmacology, British Heart Foundation Centre of Research Excellence, King's College London, 3.07 Franklin-Wilkins Building, 150 Stamford Street, London, UK.
authors: Nan
authors: N
authors: Chen
authors: Cardiovascular Division, Department of Clinical Pharmacology, British Heart Foundation Centre of Research Excellence, King's College London, 3.07 Franklin-Wilkins Building, 150 Stamford Street, London, UK.
authors: Qiuping
authors: Q
authors: Zhang
authors: Division of Asthma, Allergy, and Lung Biology, King's College London, London, UK.
authors: Yasin
authors: Y
authors: Shaifta
authors: Cardiovascular Division, Department of Clinical Pharmacology, British Heart Foundation Centre of Research Excellence, King's College London, 3.07 Franklin-Wilkins Building, 150 Stamford Street, London, UK.
authors: Genevieve
authors: G
authors: Vanderslott
authors: Cardiovascular Division, Department of Clinical Pharmacology, British Heart Foundation Centre of Research Excellence, King's College London, 3.07 Franklin-Wilkins Building, 150 Stamford Street, London, UK.
authors: Gabriella
authors: G
authors: Passacquale
authors: Cardiovascular Division, Department of Clinical Pharmacology, British Heart Foundation Centre of Research Excellence, King's College London, 3.07 Franklin-Wilkins Building, 150 Stamford Street, London, UK.
authors: Yanhua
authors: Y
authors: Hu
authors: Cardiovascular Division, Department of Clinical Pharmacology, British Heart Foundation Centre of Research Excellence, King's College London, 3.07 Franklin-Wilkins Building, 150 Stamford Street, London, UK.
authors: Qingbo
authors: Q
authors: Xu
authors: Division of Asthma, Allergy, and Lung Biology, King's College London, London, UK.
authors: Jeremy P T
authors: JP
authors: Ward
authors: Cardiovascular Division, Department of Clinical Pharmacology, British Heart Foundation Centre of Research Excellence, King's College London, 3.07 Franklin-Wilkins Building, 150 Stamford Street, London, UK albert.ferro@kcl.ac.uk.
authors: Albert
authors: A
authors: Ferro
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D004789:Enzyme Activation; D020536:Enzyme Activators / Q000494:pharmacology; D038362:Glycogen Synthase Kinase 3; D000071679:Glycogen Synthase Kinase 3 beta; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D008810:Mice, Inbred C57BL; D018919:Neovascularization, Physiologic / Q000187:drug effects; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D058986:Phosphodiesterase 5 Inhibitors / Q000494:pharmacology; D010766:Phosphorylation; D011485:Protein Binding; D012694:Serine; D015398:Signal Transduction; D051176:beta Catenin / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
chemical_list: C495270:CTNNB1 protein, human; C495267:CTNNB1 protein, mouse; D020536:Enzyme Activators; D020030:Nitric Oxide Donors; D058986:Phosphodiesterase 5 Inhibitors; D051176:beta Catenin; D009569:Nitric Oxide; D012694:Serine; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse; D000071679:Glycogen Synthase Kinase 3 beta; D017869:Cyclic GMP-Dependent Protein Kinases; D038362:Glycogen Synthase Kinase 3; D006152:Cyclic GMP
keywords: Angiogenesis; Beta catenin; Gene transcription; Nitric oxide