Data from Regulation of Vascular Endothelial Growth Factor D by Orphan Receptors Hepatocyte Nuclear Factor-4α and Chicken Ovalbumin Upstream Promoter Transcription Factors 1 and 2

Abstract

<div>Abstract<p>Vascular endothelial growth factor D has recently been linked to the control of lymphangiogenesis and lymphatic metastasis. The molecular determinants regulating <i>vegf-D</i> gene transcription, however, have not yet been identified. After isolation of 2 kb of 5′-flanking DNA of the human <i>vegf-D</i> gene, we identified a novel, atypical direct repeat (DR) element consisting of a consensus half-site (AGGTCA) at −125/−119 and a degenerated DR half-site (ATGTTA) at −99/−94 as sufficient and necessary for <i>vegf-D</i> transcription. The <i>vegf-D</i> DR element is bound and activated by the orphan receptors hepatocyte nuclear factor 4α (HNF-4α) and chicken ovalbumin upstream promoter transcription factor (COUP-TF)-1/COUP-TF2. Additionally, chromatin immunoprecipitation assays identified transcriptional coactivators cyclic AMP–responsive element binding protein–binding protein and glucocorticoid receptor interacting protein 1 at the <i>vegf-D</i> DR element and functional assays confirmed their stimulatory effect on the <i>vegf-D</i> promoter. Histone deacetylase inhibition by trichostatin A led to accumulation of acetylated histones H3/H4 at the <i>vegf-D</i> promoter, up-regulation of <i>vegf-D</i> mRNA levels, and transactivation of <i>vegf-D</i> promoter reporter gene constructs in cancer cell lines. This study for the first time describes the molecular determinants in <i>cis</i> and <i>trans</i> controlling <i>vegf-D gene</i> transcription and identifies interaction of HNF-4α and COUP-TF1/COUP-TF2 with a proximal, atypical DR element as indispensable for <i>vegf-D</i> transcription. Moreover, our findings suggest that epigenetic control of histone acetylation represents an important determinant of <i>vegf-D</i> gene expression in cancer cells. These results provide novel insights into the molecular machinery controlling <i>vegf-D</i> gene expression and may add to a better understanding of the regulation of lymphangiogenesis in vascular development and cancer. [Cancer Res 2008;68(2):457–66]</p></div>