Role of Hypoxia-Inducible Factor (HIF)-1α versus HIF-2α in the Regulation of HIF Target Genes in Response to Hypoxia, Insulin-Like Growth Factor-I, or Loss of von Hippel-Lindau Function: Implications for Targeting the HIF Pathway

Publisher Website
Google Scholar
Abstract
Overexpression of hypoxia-inducible factors (HIF), HIF-1α and HIF-2α, leads to the up-regulation of genes involved in proliferation, angiogenesis, and glucose metabolism and is associated with tumor progression in several cancers. However, the contribution of HIF-1α versus HIF-2α to vascular endothelial growth factor (VEGF) expression and other HIF-regulated target genes under different conditions is unclear. To address this, we used small interfering RNA (siRNA) techniques to knockdown HIF-1α and/or HIF-2α expression in response to hypoxia, insulin-like growth factor (IGF)-I, or renal carcinoma cells expressing constitutively high basal levels of HIF-1α and/or HIF-2α due to loss of von Hippel-Lindau (VHL) function. We found that HIF-1α primarily regulates transcriptional activation of VEGF in response to hypoxia and IGF-I compared with HIF-2α in MCF-7 cells. We also observed a reciprocal relationship between HIF-1α and HIF-2α expression in hypoxia in these cells: HIF-2α siRNA enhanced HIF-1α–mediated VEGF expression in MCF-7 cells in response to hypoxia, which could be completely blocked by cotransfection with HIF-1α siRNA. In contrast, in renal carcinoma cells that constitutively express HIF-1α and HIF-2α due to loss of VHL function, we found that high basal VEGF, glucose transporter-1, urokinase-type plasminogen activator receptor, and plasminogen activator inhibitor-1 expression was predominantly dependent on HIF-2α. Finally, we showed that a newly identified small-molecule inhibitor of HIF-1, NSC-134754, is also able to significantly decrease HIF-2α protein expression and HIF-2α–regulated VEGF levels in renal carcinoma cells. Our data have important implications for how we target the HIF pathway therapeutically. (Cancer Res 2006; 66(12): 6264-70)