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Papers In Press, published online ahead of print June 28, 2006
Cancer Research-UK labs & Section of Cancer Cell Biology, Imperial College London, London, London W12 0NN
Corresponding Author: eric.lam{at}imperial.ac.uk
In this study, we identified the forkhead transcription factor, FoxM1 as a physiological regulator of ER
J. Biol. Chem, 10.1074/jbc.M603906200
Submitted on April 24, 2006
Revised on June 28, 2006
Accepted on June 28, 2006
The Forkhead box M1 protein regulates the transcription of the estrogen receptor alpha in breast cancer cells
expression in breast carcinoma cells. Our survey of a panel of 16 different breast cell lines showed a good correlation (13/16) between FoxM1 expression and expression of ER at both protein and mRNA levels. We also demonstrated that ectopic expression of FoxM1 in two different ER-positive breast cancer cell lines, MCF-7 and ZR-75-30, led to up-regulation of ER expression at protein and transcript levels. Furthermore, treatment of MCF-7 cells with the MEK inhibitor U0126, which blocks ERK1/2 dependent-activation of FoxM1, also repressed ER expression. Consistent with this, silencing of FoxM1 expression in MCF-7 cells using siRNA resulted in almost complete abrogation of ER expression. We also went on to show that FoxM1 can activate the transcriptional activity of human ER promoter primarily through two closely located Forkhead Response Elements (FHREs) located at the proximal region of the ERpromoter. Chromatin immunoprecipitation and biotinylated oligonucleotides pull-down assays have allowed us to confirm these FHREs as important for FoxM1 binding. Further co-immunoprecipitation experiments showed that FoxO3a and FoxM1 interact in vivo. Together with the chromatin immunoprecipitation and biotinylated oligonucleotides pull-down data, the co-immunoprecipitation results also suggest the possibility that FoxM1 and FoxO3a cooperate to regulate ER gene transcription.
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