title: Progesterone receptor-cyclin D1 complexes induce cell cycle-dependent transcriptional programs in breast cancer cells.
abstract: The progesterone receptor (PR) and its coactivators are direct targets of activated cyclin-dependent kinases (CDKs) in response to peptide growth factors, progesterone, and deregulation of cell cycle inhibitors. Herein, using the T47D breast cancer model, we probed mechanisms of cell cycle-dependent PR action. In the absence of exogenous progestin, the PR is specifically phosphorylated during the G2/M phase. Accordingly, numerous PR target genes are cell cycle regulated, including HSPB8, a heat-shock protein whose high expression is associated with tamoxifen resistance. Progestin-induced HSPB8 expression required cyclin D1 and was insensitive to antiestrogens but blocked by antiprogestins or inhibition of specificity factor 1 (SP1). HSPB8 expression increased with or without ligand when cells were G2/M synchronized or contained high levels of cyclin D1. Knockdown of PRs abrogated ligand-independent HSPB8 expression in synchronized cells. Notably, PRs and cyclin D1 copurified in whole-cell lysates of transiently transfected COS-1 cells and in PR-positive T47D breast cancer cells expressing endogenous cyclin D1. PRs, cyclin D1, and SP1 were recruited to the HSPB8 promoter in progestin-treated T47D breast cancer cells. Mutation of PR Ser345 to Ala (S345A) or inhibition of CDK2 activity using roscovitine disrupted PR/cyclin D1 interactions with DNA and blocked HSPB8 mRNA expression. Interaction of phosphorylated PRs with SP1 and cyclin D1 provides a mechanism for targeting transcriptionally active PRs to selected gene promoters relevant to breast cancer progression. Understanding the functional linkage between PRs and cell cycle regulatory proteins will provide keys to targeting novel PR/cyclin D1 cross talk in both hormone-responsive disease and HSPB8-high refractory disease with high HSPB8 expression.
authors.affiliation: Departments of Medicine and Pharmacology (G.E.D., T.P.K., A.R.D., C.R.H., C.H.D., C.A.L.), Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota 55455; and Departments of Cancer Biology, Urology, and Radiation Oncology (M.J.S., K.E.K.), Kimmel Cancer Center Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
authors.forename: Gwen E
authors.initials: GE
authors.lastname: Dressing
authors.forename: Todd P
authors.initials: TP
authors.lastname: Knutson
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Schiewer
authors.forename: Andrea R
authors.initials: AR
authors.lastname: Daniel
authors.forename: Christy R
authors.initials: CR
authors.lastname: Hagan
authors.forename: Caroline H
authors.initials: CH
authors.lastname: Diep
authors.forename: Karen E
authors.initials: KE
authors.lastname: Knudsen
authors.forename: Carol A
authors.initials: CA
authors.lastname: Lange
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology*; D019556:COS Cells; D002453:Cell Cycle / Q000235:genetics*; D002457:Cell Extracts; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D019938:Cyclin D1 / Q000378:metabolism*; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006360:Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D018832:Molecular Chaperones; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011980:Receptors, Progesterone / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D002457:Cell Extracts; C413214:HSPB8 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011980:Receptors, Progesterone; D019938:Cyclin D1; D010768:Phosphoserine; D017346:Protein Serine-Threonine Kinases; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2