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J. Lipid Res.
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Journal of Lipid Research, Vol. 43, 383-391, March 2002
Copyright © 2002 by Lipid Research, Inc.

Estrogen receptor-mediated repression of human hepatic lipase gene transcription

Daniel R. Jonesa, Robert J. Schmidta, Richard T. Pickarda, Patricia S. Foxworthya, and Patrick I. Eachoa
a Lilly Research Laboratories, Cardiovascular Research Division, Eli Lilly and Company, Indianapolis, IN 46285

Correspondence to: Patrick I. Eacho, To whom correspondence should be addressed., eacho{at}lilly.com (E-mail)

Estrogen replacement therapy in women decreases hepatic lipase (HL) activity, which may account for the associated increase in HDL cholesterol. To investigate whether estrogen decreases HL transcription, transient cotransfection assays with HL promoter and estrogen receptor-{alpha} (ER{alpha}) expression constructs were performed in HepG2 cells. 17ß-estradiol (E2) decreased transcription driven by the -1557/+41 human HL promoter by up to 50% at 10-7 M. Mutation of ER{alpha} by deletion of its transactivation domains or ligand-binding domain eliminated E2-induced repression of the promoter, whereas deletion of the DNA-binding domain of ER{alpha} resulted in a 7-fold activation by E2. The E2-induced repression was maintained after mutation of a potential estrogen-response element in the promoter. The region of estrogen responsiveness was localized to -1557/-1175 of the HL promoter by deletion analysis. Mutation of an AP-1 site at -1493 resulted in a partial loss of E2-induced repression, similar to that caused by deletion of nucleotides -1557 to -1366. Gel shift assays with nuclear extracts from E2-treated HepG2 cells stably expressing ER{alpha} demonstrated an increase in binding to an AP-1 consensus oligonucleotide. The AP-1 activator, phorbol 12-myristate 13-acetate, inhibited the HL promoter by greater than 50%.

Collectively, the data suggest that estrogen represses the transcription of the HL gene, possibly through an AP-1 pathway. — Jones, D. R., R. J. Schmidt, R. T. Pickard, P. S. Foxworthy, and P. I. Eacho. Estrogen receptor-mediated repression of human hepatic lipase gene transcription. J. Lipid Res. 2002. 43: 383–391.

Supplementary key words: HepG2 cells, AP-1, phorbol ester, very low density apolipoprotein II, promoter regulation, 17ß-estradiol


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