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2‐oxoglutarate downregulates expression of vascular endothelial growth factor and erythropoietin through decreasing hypoxia‐inducible factor‐1α and inhibits angiogenesis
Author(s) -
Matsumoto Ken,
Imagawa Shigehiko,
Obara Naoshi,
Suzuki Norio,
Takahashi Satoru,
Nagasawa Toshiro,
Yamamoto Masayuki
Publication year - 2006
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.20733
Subject(s) - mg132 , erythropoietin , angiogenesis , vascular endothelial growth factor , hypoxia inducible factors , biology , hif1a , western blot , microbiology and biotechnology , ubiquitin ligase , chemistry , ubiquitin , biochemistry , proteasome , cancer research , proteasome inhibitor , vegf receptors , endocrinology , gene
In oxygenated cells, hypoxia‐inducible factor‐1 (HIF‐1) α subunits are rapidly degraded by a mechanism that involves ubiquitination by the von Hippel–Lindau tumor suppressor E3 ligase complex using 2‐oxoglutarate as a substrate. We examined the effect of 2‐oxoglutarate on the production of erythropoietin and vascular endothelial growth factor (VEGF). The expression of erythropoietin and VEGF protein were dose‐dependently downregulated in Hep3B cells by the addition of 2‐oxoglutarate. The promoter activity of VEGF‐luciferase was dose‐dependently downregulated by the addition of 2‐oxoglutarate. Gel mobility shift assays revealed that the addition of 2‐oxoglutarate dose‐dependently inhibited HIF‐1 binding activity, but did not affect GATA binding activity. Western blot analysis revealed that 2‐oxoglutarate dose‐dependently inhibited the HIF‐1α protein level in Hep3B cells in hypoxic conditions. However, MG132 (the proteasome inhibitor) rescued the inhibition of HIF‐1α protein expression by 2‐oxoglutarate. Furthermore, under hypoxic conditions, 2‐oxoglutarate dose‐dependently inhibited tube formation in in vitro angiogenesis assays. These results indicate that 2‐oxoglutarate treatment may be useful for the inhibition of angiogenesis. J. Cell. Physiol. 209: 333–340, 2006. © 2006 Wiley‐Liss, Inc.