Up-regulation of hypoxia-inducible factors HIF-1 alpha and HIF-2 alpha under normoxic conditions in renal carcinoma cells by von Hippel-Lindau tumor suppressor gene loss of function

Hypoxia induces transcription of a range of physiologically important genes including erythropoietin and vascular endothelial growth factor. The trans criptional activation is mediated by the hypoxia-inducible factor-1 (HIF-1) , a heterodimeric member of the basic helix-loop-helix PAS family, composed of alpha and beta subunits, HLF-1 alpha shares 48 per cent identity with t he recently identified HIF-2 alpha protein that is also stimulated by hypox ia, In a previous study of hemangioblastomas, the most frequent manifestati on of hereditary von Hippel-Lindau disease (VHL), we found elevated levels of vascular endothelial growth factor and HIF-2 alpha. mRNA in stromal cell s of the tumors. Mutations of the VHL tumor suppressor gene are associated with a variety of tumors such as renal clear cell carcinomas (RCC), In this study, we analysed the expression of the hypoxia-inducible factors HIF-1 a lpha and HIF-2 alpha in a range of VHL wildtype and VHL deficient RCC cell lines. In the presence of functional VHL protein, HIF-1 alpha mRNA levels a re elevated, whereas HIF-2 alpha mRNA expression is increased only in cells lacking a functional VHL gene product. On the protein levels, however, in VHL deficient cell lines, both HIF-alpha subunits are constitutively expres sed, whereas re-introduction of a functional VHL gene restores the instabil ity of HIF-1 alpha and HIF-2 alpha proteins under normoxic conditions. More over, immunohistochemical analyses of RCCs and hemangioblastomas demonstrat e up-regulation of HIF-1 alpha and HIF-2 alpha in the tumor cells. The data presented here provide evidence for a role of the VHL protein in regulatio n of angiogenesis and erythropoiesis mediated by the HIF-1 alpha and HIF-2 alpha proteins.