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dc.contributor.authorMatou-Nasri, S
dc.contributor.authorGaffney, J
dc.contributor.authorKumar, Shant
dc.contributor.authorSlevin, M
dc.date.accessioned2010-08-07T10:46:38Z
dc.date.available2010-08-07T10:46:38Z
dc.date.issued2009-10
dc.identifier.citationOligosaccharides of hyaluronan induce angiogenesis through distinct CD44 and RHAMM-mediated signalling pathways involving Cdc2 and gamma-adducin. 2009, 35 (4):761-73 Int. J. Oncol.en
dc.identifier.issn1791-2423
dc.identifier.pmid19724912
dc.identifier.urihttp://hdl.handle.net/10541/109254
dc.description.abstractWe used short-interfering RNA (siRNA) to knockdown the hyaluronan (HA) receptors CD44 and the receptor for hyaluronan-mediated motility (RHAMM) in vascular endothelial cells to investigate their role in angiogenesis. We showed that CD44 and RHAMM single knockdown inhibited low molecular weight hyaluronan (o-HA)-induced endothelial cell tube formation in Matrigel, but no change in the control, epidermal growth factor-induced tube formation was observed. Using a Kinexus phosphoprotein array and confirmational Western blotting we were able to show a differential effect on HA-induced protein expression after CD44 and RHAMM knockdown. CD44 knockdown abolished o-HA-induced membrane phospho-protein kinase C-alpha (PKC-alpha) and down-stream phospho-gamma-adducin expression. Using the PKC inhibitor Go6976, we demonstrated the involvement of PKC-alpha and gamma-adducin in o-HA-induced tube formation, whilst o-HA-induced enzymatic activity of MMP9 was also reduced. This suggests that endothelial tube formation involves activation of MMP9 via PKC-alpha. Furthermore, the involvement of gamma-adducin in o-HA-induced F-actin cytoskeleton rearrangement was CD44-dependent and the reduction of CD44 expression lead to a change in endothelial cell morphology. Both RHAMM and CD44 knockdown completely inhibited o-HA-induced Cdc2 (Cdk1) phosphorylation suggesting a possible involvement in cell cycle control. Although CD44 and RHAMM are both involved in o-HA-induced endothelial tube formation in Matrigel, they mediate distinct angiogenic signalling pathway and for the first time we demonstrated the specific involvement of gamma-adducin in CD44/o-HA-induced endothelial tube formation. The data presented here extend our understanding of key stages of the processes of o-HA-induced angiogenesis which may have relevance to tumour progression.
dc.language.isoenen
dc.subject.meshActins
dc.subject.meshAnimals
dc.subject.meshAntigens, CD44
dc.subject.meshBlotting, Western
dc.subject.meshCDC2 Protein Kinase
dc.subject.meshCalmodulin-Binding Proteins
dc.subject.meshCarbazoles
dc.subject.meshCattle
dc.subject.meshCell Shape
dc.subject.meshCells, Cultured
dc.subject.meshEndothelial Cells
dc.subject.meshEpidermal Growth Factor
dc.subject.meshExtracellular Matrix Proteins
dc.subject.meshFluorescent Antibody Technique
dc.subject.meshHyaluronic Acid
dc.subject.meshMatrix Metalloproteinase 9
dc.subject.meshNeovascularization, Physiologic
dc.subject.meshOligosaccharides
dc.subject.meshPhosphorylation
dc.subject.meshProtein Array Analysis
dc.subject.meshProtein Kinase C-alpha
dc.subject.meshProtein Kinase Inhibitors
dc.subject.meshRNA Interference
dc.subject.meshRNA, Small Interfering
dc.subject.meshReverse Transcriptase Polymerase Chain Reaction
dc.subject.meshSignal Transduction
dc.titleOligosaccharides of hyaluronan induce angiogenesis through distinct CD44 and RHAMM-mediated signalling pathways involving Cdc2 and gamma-adducin.en
dc.typeArticleen
dc.contributor.departmentSchool of Biology, Chemistry and Health Science, Manchester Metropolitan University, Manchester M1 5GD, UK.en
dc.identifier.journalInternational Journal of Oncologyen
html.description.abstractWe used short-interfering RNA (siRNA) to knockdown the hyaluronan (HA) receptors CD44 and the receptor for hyaluronan-mediated motility (RHAMM) in vascular endothelial cells to investigate their role in angiogenesis. We showed that CD44 and RHAMM single knockdown inhibited low molecular weight hyaluronan (o-HA)-induced endothelial cell tube formation in Matrigel, but no change in the control, epidermal growth factor-induced tube formation was observed. Using a Kinexus phosphoprotein array and confirmational Western blotting we were able to show a differential effect on HA-induced protein expression after CD44 and RHAMM knockdown. CD44 knockdown abolished o-HA-induced membrane phospho-protein kinase C-alpha (PKC-alpha) and down-stream phospho-gamma-adducin expression. Using the PKC inhibitor Go6976, we demonstrated the involvement of PKC-alpha and gamma-adducin in o-HA-induced tube formation, whilst o-HA-induced enzymatic activity of MMP9 was also reduced. This suggests that endothelial tube formation involves activation of MMP9 via PKC-alpha. Furthermore, the involvement of gamma-adducin in o-HA-induced F-actin cytoskeleton rearrangement was CD44-dependent and the reduction of CD44 expression lead to a change in endothelial cell morphology. Both RHAMM and CD44 knockdown completely inhibited o-HA-induced Cdc2 (Cdk1) phosphorylation suggesting a possible involvement in cell cycle control. Although CD44 and RHAMM are both involved in o-HA-induced endothelial tube formation in Matrigel, they mediate distinct angiogenic signalling pathway and for the first time we demonstrated the specific involvement of gamma-adducin in CD44/o-HA-induced endothelial tube formation. The data presented here extend our understanding of key stages of the processes of o-HA-induced angiogenesis which may have relevance to tumour progression.


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