Estrogen regulation of vascular endothelial growth factor gene expression in ZR-75 breast cancer cells through interaction of estrogen receptor alpha and SP proteins

Oncogene. 2004 Feb 5;23(5):1052-63. doi: 10.1038/sj.onc.1207201.


Vascular endothelial growth factor (VEGF) is expressed in multiple hormone-dependent cancer cells/tumors. Treatment of ZR-75 breast cancer cells with 17beta-estradiol (E2) induced a greater than fourfold increase of VEGF mRNA levels. ZR-75 breast cancer cells were transfected with pVEGF1, a construct containing a -2018 to +50 VEGF promoter insert, and E2 induced reporter gene (luciferase) activity. Deletion and mutation analysis of the VEGF gene promoter identified a GC-rich region (-66 to -47) which was required for E2-induced transactivation of pVEGF5, a construct containing the minimal promoter (-66 to +54) that exhibited E2-responsiveness. Interactions of nuclear proteins from ZR-75 cells with the proximal GC-rich region of the VEGF gene promoter were investigated by electrophoretic mobility shift and chromatin immunoprecipitation assays. The results demonstrate that both Sp1 and Sp3 proteins bound the GC-rich motif (-66 to -47), and estrogen receptor alpha (ERalpha) interactions were confirmed by chromatin immunoprecipitation. Moreover, E2-dependent activation of constructs containing proximal and distal GC/GT-rich regions of the VEGF promoter was inhibited in ZR-75 cells transfected with small inhibitory RNAs for Sp1 and Sp3. These results were consistent with a mechanism of hormone activation of VEGF through ERalpha/Sp1 and ERalpha/Sp3 interactions with GC-rich motifs.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Breast Neoplasms / genetics*
  • Breast Neoplasms / metabolism
  • Cell Line, Tumor
  • Estradiol / pharmacology
  • Estrogen Receptor alpha
  • Estrogens / pharmacology*
  • GC Rich Sequence
  • Gene Deletion
  • Gene Expression Regulation, Neoplastic / drug effects*
  • Genes, Reporter
  • Humans
  • Neoplasms, Hormone-Dependent / metabolism
  • Promoter Regions, Genetic
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism
  • Receptors, Estrogen / metabolism*
  • Sp1 Transcription Factor / metabolism*
  • Transcriptional Activation
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / metabolism*


  • Estrogen Receptor alpha
  • Estrogens
  • RNA, Messenger
  • RNA, Small Interfering
  • Receptors, Estrogen
  • Sp1 Transcription Factor
  • Vascular Endothelial Growth Factor A
  • Estradiol