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Papers In Press, published online ahead of print October 7, 2002
Department of Medicine, University of California, Irvine, Long Beach, CA 90822
Corresponding Author: ellis.levin{at}med.va.gov
Estrogen binds to receptors that translocate to the plasma membrane and to the nucleus. The rapid, non-genomic actions of this sex steroid are attributed to membrane action, while gene transcription occurs through nuclear receptor function. However, gene transcription can also result from estrogen signaling initiated at the membrane but the relative importance of this mechanism is not known. In vascular endothelial cells (EC), estradiol (E2) activates several kinase cascades including PI3K/AKT, a signaling pathway that impacts EC biology. We determined here by DNA microarray that 40 minutes exposure to E2 significantly increased 250 genes in EC, upregulation that was substantially prevented by the PI3K inhibitor, LY294002. This coincided with maximum E2-induced PI3K activity at 15-30 minutes. An important vascular gene strongly upregulated by E2 in our array produces cyclooxygenase-2 (Cox-2). In cultured EC, E2 induced both Cox-2 gene expression and new Cox-2 protein synthesis by 40 and 60 minutes respectively, and rapidly stimulated the secretion of prostaglandins PGI2 and PGE2. The upregulation of gene expression reflected transcriptional transactivation, shown using Cox-2 promoter/luciferase reporters in the EC. Soluble inhibitors or dominant negative constructs for PI3K and AKT prevented all these actions of E2. Functionally, EC migration was induced by the sex steroid, and this was significantly reversed by NS-398, a Cox-2 inhibitor. Gene transcription and cell biological effects of estrogen emanate from rapid and specific signaling, integrating cell surface and nuclear actions of this steroid.
J. Biol. Chem, 10.1074/jbc.M210106200
Submitted on October 2, 2002
Revised on October 7, 2002
Accepted on October 7, 2002
Integration of the non-genomic and genomic actions of estrogen: Membrane initiated signaling by steroid (MISS) to transcription and cell biology
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