Estradiol-induced vascular endothelial growth factor-A expression in breast tumor cells is biphasic and regulated by estrogen receptor-α dependent pathway

Estrogens have been shown to regulate vascular endothelial growth factor-A (VEGF-A) for physiological and patho-physiological functions. However, estrogen action on VEGF-A mRNA expression has not been completely elucidated. We have identified two phases of activation of VEGF-A mRNA transcription, one early and one late response, induced by 17β-estradiol (17β-E2) in ER+ MCF-7 breast tumor cells, depending upon the length of exposure. VEGF-A mRNA level was significantly higher than control in tumor cells after 2 h of 17β-E2 exposure. Furthermore this induction was not inhibited by cycloheximide, indicating that it was a direct effect of estrogen. In contrast VEGF-A mRNA expression was back at basal level in MCF-7 cells exposed to 17β-E2 for 6 h. However, expression levels were again significantly augmented after 24 h of exposure, and this induction was unaltered by cycloheximide indicating that de novo protein synthesis was not required and like early response, it was a direct effect of estrogen. The antiestrogen ICI 182,780 was a pure antagonist for the early response phase of VEGF-A mRNA induction, but it had partial but significant effect on the late response phase, further suggesting that both early and late phases were ER dependent. In human mammary epithelial cells (HMEC) lacking estrogen receptor (ER-α) the early and late response phase of VEGF-A mRNA induction in response to 17β-E2 was not found, but significant inductions were seen in the early and late phases when ER-α transfected HMEC were exposed to 17β-E2 for 2 or 24 h. Taken together, these studies suggest that VEGF-A is an estrogen responsive gene and modulation of this gene expression by estrogen is biphasic and can be mediated through ER-α dependent pathway.