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Papers In Press, published online ahead of print May 20, 2002
Department of Medicine, University of Colorado Health Sciences Center, Denver, CO 80262
Corresponding Author: Britta.Jacobsen{at}uchsc.edu
All known progesterone target cells co-express two functionally different progesterone receptor (PR) isoforms: 120 kDa B-receptors (PR-B) and N-terminally truncated, 94 kDa A-receptors (PR-A). Their ratio varies in normal and malignant tissues. In human breast cancer cells, homodimers of progesterone-occupied PR-A or PR-B regulate different gene subsets. To study PR homo- and heterodimers we constructed breast cancer cell lines in which isoform expression is controlled by an inducible system. PR-negative cells, or cells that stably express one or the other isoform, were used to construct five sets of cells: i) PR-negative control cells (Y iNull); ii) inducible PR-A (Y iA); iii) inducible PR-B (Y iB); iv) stable PR-B plus inducible PR-A (B iA); and v) stable PR-A plus inducible PR-B (A iB). Expression levels of each isoform and/or the PR-A:PR-B ratios can be tightly controlled by the dose of inducer as demonstrated by immunoblot and transcription studies. Induced PR undergo normal progestin-dependent phosphorylation and downregulation, and regulate exogenous promoters as well as endogenous gene expression. Transcription of exogenous promoters is dependent on the PR-A:PR-B ratio, while transcription of endogenous genes is more complex. Finally, we describe several genes that are regulated by induced PR-A even in the absence of ligand.
J. Biol. Chem, 10.1074/jbc.M202584200
Submitted on March 18, 2002
Revised on April 30, 2002
Accepted on May 20, 2002
New human breast cancer cells to study progesterone receptor (PR) isoform ratio effects and ligand-independent gene regulation
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