Regulation of vascular endothelial growth factor (VEGF) gene transcription by estrogen receptors a and b

Abstract

Vascular endothelial growth factor (VEGF) mediates angiogenic activity in a variety of estrogen target tissues. To determine whether estrogen has a direct transcriptional effect on VEGF gene expression, we developed a model system by transiently transfecting human VEGF promoter-luciferase reporter constructs into primary human endometrial cells and into Ishikawa cells, derived from a well-differentiated human endometrial adenocarcinoma. In primary endometrial epithelial cells, treatment with 17b-estradiol (E2) resulted in a 3.8-fold increase in luciferase activity, whereas a 3.2-fold induction was demonstrated for stromal cells. Our Ishikawa cells had less than 100 functional estrogen receptors (ER)y cell and were therefore cotransfected with expression vectors encoding either the aor the b-form of the human ER. In cells cotransfected with ERa, E2 induced 3.2-fold induction in VEGFpromoter luciferase activity. A 2.3-fold increase was observed in cells cotransfected with ERb. Through specific deletions, the E2 response was restricted to a single 385-bp PvuII-SstI fragment in the 5* flanking DNA. Cotransfection of this upstream region with a DNA binding domain ER mutant, or site-directed mutagenesis of a variant ERE within this fragment, resulted in the loss of the E2 response. Electromobility shift assays demonstrated that this same ERE sequence specifically binds estradiol-ER complexes. These studies demonstrate that E2-regulated VEGF gene transcription requires a variant ERE located 1.5 kb upstream from the transcriptional start site. Site-directed mutagenesis of this ERE abrogated E2-induced VEGF gene expression.

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@inproceedings{Mller2000RegulationOV, title={Regulation of vascular endothelial growth factor (VEGF) gene transcription by estrogen receptors a and b}, author={Michael M{\"{u}ller and J Vigne and Alexander G. Minchenko and Dan I. Lebovic and Dale C. Leitman and Robert N. Taylor}, year={2000} }