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Endometrial Endothelial Cell Steroid Receptor Expression and Steroid Effects on Gene Expression

Department of Obstetrics, Gynecology, and Reproductive Sciences (G.K., F.S., J.H., C.J.L.), Yale University School of Medicine, New Haven, Connecticut 06520-8063; Center for Reproductive Sciences (R.T.), University of California, San Francisco, California 94143; Reproductive and Developmental Sciences (H.O.D.C.), Centre for Reproductive Biology, University of Edinburgh, Edinburgh EH16 4SB, United Kingdom; and Centre for Women’s Health Research (P.A.W.R.), Monash University Department of Obstetrics and Gynaecology, Monash Medical Centre, Clayton, Victoria 3168, Australia

Address all correspondence and requests for reprints to: Dr. Graciela Krikun, Department of Obstetrics, Gynecology, and Reproductive Sciences, Yale University School of Medicine, 333 Cedar Street, P.O. Box 208063, New Haven, Connecticut 06520-8063. E-mail: graciela.krikun{at}yale.edu.

Controversy exists regarding the expression of specific steroid receptor proteins and mRNA in human microvascular endometrial endothelial cells (HEECs). Thus, we studied steroid receptor expression in early passaged HEEC cultures and freshly isolated HEECs. Analysis of estrogen receptor (ER) and progesterone receptor (PR) mRNA levels was carried out with real-time quantitative RT-PCR, and the repertoire of genes activated by their respective steroid ligands was assessed by mRNA microarray analyses of 18,400 genes and expressed sequence tags. We observed that cultured and freshly isolated HEECs each express ER-ß mRNA but not ER-. In addition, PR mRNA was also detectable in both HEEC sources. Microarray analysis demonstrated that treatment of HEEC cultures with either estradiol or medroxyprogesterone acetate produced differential effects on a wide variety of genes, and cluster analysis demonstrated that many of the genes are involved in intracellular signaling and enzymatic pathways. Thus, quantitative RT-PCR and microarray analyses demonstrate that HEECs express ER-ß and PR mRNA and that gene expression by HEECs is differentially regulated by treatment with estrogen or progestin.

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