Evidence of hypoxia-inducible factor-1 in vascular endothelial and smooth muscle cells

The hypoxia-inducible element (HIE-1), a 50-bp region just 3' to the human erythropoietin gene, has been found to regulate transcription in cells that do not ordinarily synthesize erythropoietin. We hypothesized that the HIE-1 and associated protein factors may have a role in transcriptional regulation in hypoxic vascular tissues. Therefore, tissues of vascular origin were grown in culture and exposed to hypoxia (1% 02, 5% CO2, balance N2) or normoxia (21% O2, 5% CO2). Human microvascular endothelial cells (HMEC-1) studied with electrophoretic mobility shifting demonstrated that HIE-1 was bound to a protein induced by hypoxia in these cells. HMEC-1 and rat aortic smooth muscle cells (RASM) were transfected with the vector pGL2-HIE-1. HMEC-1 reporter gene expression was 3.7 +/- 0.5-fold increased at 12 hours and 3.7 +/- 1.3-fold increased at 24 hours by hypoxia. RASM reporter gene expression was 18.6 +/- 6.5-fold (SEM) increased at 12 hours and 2.0 +/- 0.7-fold increased at 24 hours by hypoxia. These findings provide indirect evidence of a hypoxia-inducible factor in vascular cellular transcriptional regulation.