December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Molecular Evidence of 17beta-Hydroxysteroid Dehydrogenases, type 2 and 7, Gene Expression in the Human Neuroendocrine Ciliary Epithelium: Synthesis and Oxidative Metabolism of Estrogens in Normal- and in Glaucoma-derived Cell Lines
Author Affiliations & Notes
  • M Coca-Prados
    Ophthalmology & Visual Sci Yale University Med School New Haven CT
  • Y Wang
    Ophthalmology and Visual Science Yale University School of Medicine New Haven CT
  • A Herrala
    Biocenter Oulu and Research Center for Molecular Endocrinology University of Oulu Oulu Finland
  • PT Vihko
    Biocenter Oulu and Research Center for Molecular Endocrinology University of Oulu Oulu Finland
  • S Ghosh
    Ophthalmology and Visual Science Yale University School of Medicine New Haven CT
  • Footnotes
    Commercial Relationships   M. Coca-Prados, None; Y. Wang, None; A. Herrala, None; P.T. Vihko, None; S. Ghosh, None. Grant Identification: EY04873
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 4065. doi:
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      M Coca-Prados, Y Wang, A Herrala, PT Vihko, S Ghosh; Molecular Evidence of 17beta-Hydroxysteroid Dehydrogenases, type 2 and 7, Gene Expression in the Human Neuroendocrine Ciliary Epithelium: Synthesis and Oxidative Metabolism of Estrogens in Normal- and in Glaucoma-derived Cell Lines . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4065.

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Abstract

Abstract: : Purpose:To identify biologically significant genes in the human nonpigmented (NPE) ciliary epithelium where their expression may be up or down regulated in glaucoma Methods:Suppression subtractive hybridization (SSH) was applied between two populations of mRNA prepared from two NPE cell lines (ODM-C4 and GCE) established in vitro from two eye donors: one normal and another with chronic open glaucoma, respectively. Approximately 100 SHH clones were isolated and identified. One clone, SSH-30, exhibited 100% identity with the human 17ß-hydroxysteroid dehydrogenase type 7 (17HSD7) cDNA and studied further. Reductive and oxidative reactions mediated by 17HSD7 and 17HSD2 respectively were determined in cultured ODM-C4 and GCE cells. Results: 17HDS7 catalyzes the interconvertion of estrone (E1) to estradiol (E2); and 17HSD2 catalyzes the interconvertion between E2 and E1. Cultured ODM-C4 cells co-expressed 17HDS7 and 17HSD2 mRNAs and their expression was up-regulated by 17ß-estradiol (10-7M) treatment in a time-dependent manner. In contrast, cultured GCE cells did not alter 17HSD2 expression in response to estradiol treatment. ODM-C4 cells but not GCE cells, exhibited a strong oxidative activity. The ODM-C4 cells were able to interconvert E2 into E1; testosterone (T) to androstendione (A-dione); and dihydrotestosterone (DHT) into 5α-androstanedione (5α-Androstane or epiandrosterone (Epiand). A low reductive activity, was detected in GCE cells. These cells were able to interconvert dihydrotestosterone (DHT) into 5α-androstanedione (5α-Androstane or epiandrosterone (Epiand). Using RT-PCR we further demonstrated that cultured ODM-C4 cells and intact human ciliary epithelium expressed estrogen receptors ER1 and ER2, cytochrome P450 CYP1B1 and CYP19, and the sex hormone-binding globulin. Conclusion:These results support the finding that the human ciliary NPE epithelial cells are a target of sex hormones, and demonstrate that they display intracrine steroidogenic activity. These studies provide new evidence of the possible role of 17HSD enzymes as key factors regulating highly active ligands for sex-steroid receptors in the endocrine functions of the human NPE cells, in normal and in glaucoma cells. CR: N Support: NIH/NEI grant EY04873.

Keywords: 417 gene/expression • 348 ciliary body • 399 enzymes/enzyme inhibitors 
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