Identification of functional estrogen response elements in the gene coding for the potent angiogenic factor vascular endothelial growth factor

Salman M. Hyder, Zafar Nawaz, Constance Chiappetta, George M. Stancel

Research output: Contribution to journalArticle

240 Citations (Scopus)

Abstract

Vascular endothelial growth factor (VEGF) is a potent stimulator of angiogenesis and a prognostic factor for many tumors including those of endocrine-responsive tissues such as the breast and uterus. We and others have previously shown that VEGF is regulated by estradiol and tamoxifen in the uterus and by estradiol in human breast cancer cells, and pharmacological evidence has suggested that this regulation was mediated by transcriptional activation of the estrogen receptor (ER). This prompted us to investigate whether the VEGF gene contains sequences that bind the ER and confer hormonal inducibility to reporter constructs in the presence of the two ER subtypes. These studies identified two sequences homologous to the consensus estrogen response element, GGTCAnnnTGACC, which bind both ER-α and ER-β. One of these elements is located in the 5'-untranslated region of the VEGF gene (GGGCAaagTGACT), and the other is located in the 3'-untranslated region (GAGCAcccTGCCC). Competition with excess unlabeled oligonucleotides indicates that these two elements bind both ERs specifically, mutations in either half- site of the two elements abolish receptor binding, and ER-α- and ER- βspecific antibodies interact with complexes formed with the corresponding receptor subtypes. In cells containing either ER-α or ER- β, the 3'element behaves as a traditional enhancer that confers hormone inducibility to reporter constructs in an orientation-independent manner, and transcriptional activity is blocked by the pure antiestrogen ICI 182,780. The pattern of transcriptional activity of the element located in the 5'-flanking region is more complex. In the orientation found in the endogenous gene, this element is nonresponsive to ER-β but confers estrogen-dependent inhibition of transcription with ER-α that is blunted by ICI 182,780. In the opposite orientation, the 5'-element confers hormone inducibility with either ER-α or -β and ICI 182,780 blocks activation by ER-α but not by ER-β. These findings support the hypotheses that estrogens directly regulate VEGF transcription in target tissues and tumors, although such regulation appears likely to involve a complex interplay of cis- and trans-acting elements not previously observed for other hormone-responsive genes.

Original languageEnglish
Pages (from-to)3183-3190
Number of pages8
JournalCancer Research
Volume60
Issue number12
StatePublished - Jun 15 2000
Externally publishedYes

Fingerprint

Angiogenesis Inducing Agents
Response Elements
Estrogen Receptors
Vascular Endothelial Growth Factor A
Estrogens
Genes
Hormones
Uterus
Estradiol
Estrogen Receptor Modulators
5' Flanking Region
5' Untranslated Regions
3' Untranslated Regions
Tamoxifen
Sequence Homology
Oligonucleotides
Transcriptional Activation
Neoplasms
Breast

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Identification of functional estrogen response elements in the gene coding for the potent angiogenic factor vascular endothelial growth factor. / Hyder, Salman M.; Nawaz, Zafar; Chiappetta, Constance; Stancel, George M.

In: Cancer Research, Vol. 60, No. 12, 15.06.2000, p. 3183-3190.

Research output: Contribution to journalArticle

@article{bfa3185d34aa41688b617a5274beae30,
title = "Identification of functional estrogen response elements in the gene coding for the potent angiogenic factor vascular endothelial growth factor",
abstract = "Vascular endothelial growth factor (VEGF) is a potent stimulator of angiogenesis and a prognostic factor for many tumors including those of endocrine-responsive tissues such as the breast and uterus. We and others have previously shown that VEGF is regulated by estradiol and tamoxifen in the uterus and by estradiol in human breast cancer cells, and pharmacological evidence has suggested that this regulation was mediated by transcriptional activation of the estrogen receptor (ER). This prompted us to investigate whether the VEGF gene contains sequences that bind the ER and confer hormonal inducibility to reporter constructs in the presence of the two ER subtypes. These studies identified two sequences homologous to the consensus estrogen response element, GGTCAnnnTGACC, which bind both ER-α and ER-β. One of these elements is located in the 5'-untranslated region of the VEGF gene (GGGCAaagTGACT), and the other is located in the 3'-untranslated region (GAGCAcccTGCCC). Competition with excess unlabeled oligonucleotides indicates that these two elements bind both ERs specifically, mutations in either half- site of the two elements abolish receptor binding, and ER-α- and ER- βspecific antibodies interact with complexes formed with the corresponding receptor subtypes. In cells containing either ER-α or ER- β, the 3'element behaves as a traditional enhancer that confers hormone inducibility to reporter constructs in an orientation-independent manner, and transcriptional activity is blocked by the pure antiestrogen ICI 182,780. The pattern of transcriptional activity of the element located in the 5'-flanking region is more complex. In the orientation found in the endogenous gene, this element is nonresponsive to ER-β but confers estrogen-dependent inhibition of transcription with ER-α that is blunted by ICI 182,780. In the opposite orientation, the 5'-element confers hormone inducibility with either ER-α or -β and ICI 182,780 blocks activation by ER-α but not by ER-β. These findings support the hypotheses that estrogens directly regulate VEGF transcription in target tissues and tumors, although such regulation appears likely to involve a complex interplay of cis- and trans-acting elements not previously observed for other hormone-responsive genes.",
author = "Hyder, {Salman M.} and Zafar Nawaz and Constance Chiappetta and Stancel, {George M.}",
year = "2000",
month = "6",
day = "15",
language = "English",
volume = "60",
pages = "3183--3190",
journal = "Journal of Cancer Research",
issn = "0099-7013",
publisher = "American Association for Cancer Research Inc.",
number = "12",

}

TY - JOUR

T1 - Identification of functional estrogen response elements in the gene coding for the potent angiogenic factor vascular endothelial growth factor

AU - Hyder, Salman M.

AU - Nawaz, Zafar

AU - Chiappetta, Constance

AU - Stancel, George M.

PY - 2000/6/15

Y1 - 2000/6/15

N2 - Vascular endothelial growth factor (VEGF) is a potent stimulator of angiogenesis and a prognostic factor for many tumors including those of endocrine-responsive tissues such as the breast and uterus. We and others have previously shown that VEGF is regulated by estradiol and tamoxifen in the uterus and by estradiol in human breast cancer cells, and pharmacological evidence has suggested that this regulation was mediated by transcriptional activation of the estrogen receptor (ER). This prompted us to investigate whether the VEGF gene contains sequences that bind the ER and confer hormonal inducibility to reporter constructs in the presence of the two ER subtypes. These studies identified two sequences homologous to the consensus estrogen response element, GGTCAnnnTGACC, which bind both ER-α and ER-β. One of these elements is located in the 5'-untranslated region of the VEGF gene (GGGCAaagTGACT), and the other is located in the 3'-untranslated region (GAGCAcccTGCCC). Competition with excess unlabeled oligonucleotides indicates that these two elements bind both ERs specifically, mutations in either half- site of the two elements abolish receptor binding, and ER-α- and ER- βspecific antibodies interact with complexes formed with the corresponding receptor subtypes. In cells containing either ER-α or ER- β, the 3'element behaves as a traditional enhancer that confers hormone inducibility to reporter constructs in an orientation-independent manner, and transcriptional activity is blocked by the pure antiestrogen ICI 182,780. The pattern of transcriptional activity of the element located in the 5'-flanking region is more complex. In the orientation found in the endogenous gene, this element is nonresponsive to ER-β but confers estrogen-dependent inhibition of transcription with ER-α that is blunted by ICI 182,780. In the opposite orientation, the 5'-element confers hormone inducibility with either ER-α or -β and ICI 182,780 blocks activation by ER-α but not by ER-β. These findings support the hypotheses that estrogens directly regulate VEGF transcription in target tissues and tumors, although such regulation appears likely to involve a complex interplay of cis- and trans-acting elements not previously observed for other hormone-responsive genes.

AB - Vascular endothelial growth factor (VEGF) is a potent stimulator of angiogenesis and a prognostic factor for many tumors including those of endocrine-responsive tissues such as the breast and uterus. We and others have previously shown that VEGF is regulated by estradiol and tamoxifen in the uterus and by estradiol in human breast cancer cells, and pharmacological evidence has suggested that this regulation was mediated by transcriptional activation of the estrogen receptor (ER). This prompted us to investigate whether the VEGF gene contains sequences that bind the ER and confer hormonal inducibility to reporter constructs in the presence of the two ER subtypes. These studies identified two sequences homologous to the consensus estrogen response element, GGTCAnnnTGACC, which bind both ER-α and ER-β. One of these elements is located in the 5'-untranslated region of the VEGF gene (GGGCAaagTGACT), and the other is located in the 3'-untranslated region (GAGCAcccTGCCC). Competition with excess unlabeled oligonucleotides indicates that these two elements bind both ERs specifically, mutations in either half- site of the two elements abolish receptor binding, and ER-α- and ER- βspecific antibodies interact with complexes formed with the corresponding receptor subtypes. In cells containing either ER-α or ER- β, the 3'element behaves as a traditional enhancer that confers hormone inducibility to reporter constructs in an orientation-independent manner, and transcriptional activity is blocked by the pure antiestrogen ICI 182,780. The pattern of transcriptional activity of the element located in the 5'-flanking region is more complex. In the orientation found in the endogenous gene, this element is nonresponsive to ER-β but confers estrogen-dependent inhibition of transcription with ER-α that is blunted by ICI 182,780. In the opposite orientation, the 5'-element confers hormone inducibility with either ER-α or -β and ICI 182,780 blocks activation by ER-α but not by ER-β. These findings support the hypotheses that estrogens directly regulate VEGF transcription in target tissues and tumors, although such regulation appears likely to involve a complex interplay of cis- and trans-acting elements not previously observed for other hormone-responsive genes.

UR - http://www.scopus.com/inward/record.url?scp=0034086611&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034086611&partnerID=8YFLogxK

M3 - Article

VL - 60

SP - 3183

EP - 3190

JO - Journal of Cancer Research

JF - Journal of Cancer Research

SN - 0099-7013

IS - 12

ER -