Hypoxia induces vascular endothelial growth factor gene transcription in human osteoblast-like cells through the hypoxia-inducible factor-2alpha.

Abstract:

:VEGF is produced by osteoblasts and has been postulated to function as an angiogenic stimulus during normal skeletal development and in fracture repair. In this study, we characterized the molecular mechanisms by which experimental hypoxia increases VEGF mRNA in human MG63 osteoblast-like cells. Exposure of MG63 cells to 1% O(2) for 24 h resulted in a four-fold increase in VEGF mRNA. Immunoblotting of nuclear extracts demonstrated a time-dependent increase in the level of the Hif-2alpha protein, which preceded the rise in VEGF mRNA. To determine the effect of hypoxia on VEGF gene transcription, MG63 cells were transiently transfected with a segment of the VEGF promoter construct fused to luciferase and then exposed to 1% O(2). Hypoxia induced VEGF promoter activity five-fold by 24 h. Forced expression of Hif-2alpha, but not Hif-1alpha, increased both basal and hypoxia induced VEGF promoter activity. By contrast, the ability of the VEGF reporter to respond to hypoxia or recombinant Hif-2alpha was abolished in cells transfected with a VEGF promoter construct containing a mutation in the hypoxia response element. In summary, exposure of osteoblast-like cells to hypoxia induces VEGF expression via induction of Hif-2alpha and transcriptional activation of the VEGF promoter.

journal_name

Endocrinology

journal_title

Endocrinology

authors

Akeno N,Czyzyk-Krzeska MF,Gross TS,Clemens TL

doi

10.1210/endo.142.2.8112

subject

Has Abstract

pub_date

2001-02-01 00:00:00

pages

959-62

issue

2

eissn

0013-7227

issn

1945-7170

journal_volume

142

pub_type

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