title: Glucose-activated RUNX2 phosphorylation promotes endothelial cell proliferation and an angiogenic phenotype.
abstract: The runt-related protein-2 (RUNX2) is a DNA-binding transcription factor that regulates bone formation, tumor cell metastasis, endothelial cell (EC) proliferation, and angiogenesis. RUNX2 DNA binding is glucose and cell cycle regulated. We propose that glucose may activate RUNX2 through changes in post-translational phosphorylation that are cell cycle-specific and will regulate EC function. Glucose increased cell cycle progression in EC through both G2/M and G1 phases with entry into S-phase occurring only in subconfluent cells. In the absence of nutrients and growth factors (starvation), subconfluent EC were delayed in G1 when RUNX2 expression was reduced. RUNX2 phosphorylation, activation of DNA binding, and pRb phosphorylation were stimulated by glucose and were necessary to promote cell cycle progression. Glucose increased RUNX2 localization at focal subnuclear sites, which co-incided with RUNX2 occupancy of the cyclin-dependent kinase (cdk) inhibitor p21(Cip1) promoter, a gene normally repressed by RUNX2. Mutation of the RUNX2 cdk phosphorylation site in the C-terminal domain (S451A.RUNX2) reduced RUNX2 phosphorylation and DNA binding. Expression of this cdk site mutant in EC inhibited glucose-stimulated differentiation (in vitro tube formation), monolayer wound healing, and proliferation. These results define a novel relationship between glucose-activated RUNX2 phosphorylation, cell cycle progression, and EC differentiation. These data suggest that inhibition of RUNX2 expression or DNA binding may be a useful strategy to inhibit EC proliferation in tumor angiogenesis.
authors.affiliation: The Graduate Program in Life Sciences, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA.
authors.forename: Adam D
authors.initials: AD
authors.lastname: Pierce
authors.forename: Ian E
authors.initials: IE
authors.lastname: Anglin
authors.forename: Michele I
authors.initials: MI
authors.lastname: Vitolo
authors.forename: Maria T
authors.initials: MT
authors.lastname: Mochin
authors.forename: Karen F
authors.initials: KF
authors.lastname: Underwood
authors.forename: Simeon E
authors.initials: SE
authors.lastname: Goldblum
authors.forename: Sravya
authors.initials: S
authors.lastname: Kommineni
authors.forename: Antonino
authors.initials: A
authors.lastname: Passaniti
mesh_terms: D002453:Cell Cycle / Q000502:physiology; D002455:Cell Division; D049109:Cell Proliferation; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism*; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D042783:Endothelial Cells / Q000502:physiology*; D005786:Gene Expression Regulation; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D009369:Neoplasms / Q000378:metabolism; D018919:Neovascularization, Physiologic; D010766:Phosphorylation; D011401:Promoter Regions, Genetic
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D050759:Cyclin-Dependent Kinase Inhibitor p21; D004268:DNA-Binding Proteins; C493709:RUNX2 protein, human; D005947:Glucose