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- W2609019349 abstract "116 The UDP-glucuronosyltransferase 2B28 (UGT2B28) gene is a member of the UGT2B subfamily whose members actively metabolize androgens and estrogens in steroid target tissue. The expression of UGT2B28 in an established breast cancer cell line suggests a role of this gene in androgen and estrogen metabolism in this tissue. UGT2B28 is composed of three different cDNA types: type I, which encodes the full cDNA, type II, which has a deletion of 308 bp in the cofactor-binding domain; type III, which lacks 351 bp in the putative substrate-binding domain. Numerous half-GRE and AP-1 sites are present in the 5′ flanking region of UGT2B28 that suggest transcriptional control of the gene by epidermal growth factor (EGF) and glucocorticoid receptor (GR). To determine the role of EGF and GR on UGT2B28 regulation, we measured the expression of endogenous UGT2B28 and performed transient transfections of luciferase reporter constructs containing various lengths of the UGT2B28 promoter region; both in LNCaP cells with and without dexamethasone (dex), a glucocorticoid mimicking compound, and EGF. The endogenous expression of UGT2B28 type I, type II and type III in LNCaP cells was measured using RT-PCR after the treatment of the cells with dex (10-7 M), EGF (20 ng/ml) and dex plus EGF. Results observed using gel electrophoresis showed a marked increase in expression for all three UGT2B28 cDNAs as a result of treatment with EGF and EGF plus dex. However, type I UGT2B28 showed stronger expression. Results from transfection with the 3 kb construct show that there is a 2.4-fold increase in transcriptional activity of LNCaP cells treated EGF when compared to controls while treatment with EGF plus dex resulted in a 50% reduction in transcriptional activity. In addition, western blot analysis has shown that LNCaP cells express EGFR, GRα, AR, NF1, and Brg1 proteins. These results suggest that EGF/EGFR mediates an increase in transcription through a pathway that may have cross-reactivity with GR signaling. Changes in UGT2B28 expression in steroid target tissue by increased EGFR signaling decrease cellular steroid concentration through increased metabolism and thereby play a role in the development of hormone-refractory cancers. This research was supported by funding from NIGMS/NIH S06-GM08049, CA92077-02, CA92075-02 and LMC/NIEHS/NIH." @default.
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- W2609019349 date "2006-04-15" @default.
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- W2609019349 title "The RheoSwitch system for inducible up- and down-regulation of gene expression" @default.
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