Hypoxia-inducible factor-1α and -2α additively promote endothelial vasculogenic properties

Abstract

Objective: While both play a role in the transcriptional response of hypoxic endothelial cells (ECs), hypoxia-inducible factor-1α (HIF-1α) and HIF-2α differ in their transactivation sites, pointing at potentially different target genes. We studied the discrete and common effects of HIF-1α and HIF-2α on the cytokine expression and vasculogenic properties of ECs. Methods and Results: H5V and bovine aortic ECs were transfected to express HIF-1α, HIF-2α or both. Overexpression of HIF-1α or HIF-2α and, to a greater extent, cotransfection of HIF-1α and HIF-2α resulted in EC activation, as revealed by analysis of the adhesion capacities and adhesion molecule surface expression of ECs. From the paracrine aspect, conditioned medium from HIF-expressing ECs was found to promote the migration and tube formation capacity of wild-type ECs, mostly following HIF-1α and HIF-2α coexpression. Antibody arrays revealed altered expression of multiple cytokines, pointing at consistent additive effects of HIF-1α and HIF-2α on angiogenic protein expression. Finally, HIF-1α and HIF-2α additively promoted vessel formation in vivo, as demonstrated by a Matrigel angiogenesis assay. Conclusion: Our results further clarify the functional roles of HIF-1α and HIF-2α in ECs and for the first time demonstrate a common contribution of HIF-1α and HIF-2α to vasculogenesis. © 2009 S. Karger AG, Basel.