Cancer Research Aziza Shad Protein Translation and Cancer
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online
Cancer Research 68, 301, January 1, 2008. doi: 10.1158/0008-5472.CAN-07-1983
© 2008 American Association for Cancer Research
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Supplementary Data
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Frietze, S.
Right arrow Articles by Brown, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Frietze, S.
Right arrow Articles by Brown, M.

Endocrinology

CARM1 Regulates Estrogen-Stimulated Breast Cancer Growth through Up-regulation of E2F1

Seth Frietze1, Mathieu Lupien2,3, Pamela A. Silver1 and Myles Brown2,3

1 Department of Systems Biology, Harvard Medical School; 2 Division of Molecular and Cellular Oncology, Department of Medical Oncology, Dana-Farber Cancer Institute; and 3 Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts

Requests for reprints: Myles Brown, Division of Molecular and Cellular Oncology, Dana-Farber Cancer Institute, 44 Binney Street, Boston, MA 02115. Phone: 617-632-3948; Fax: 617-582-8501; E-mail: myles_brown{at}dfci.harvard.edu.


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
Estrogen receptor {alpha} (ER{alpha}) mediates breast cancer proliferation through transcriptional mechanisms involving the recruitment of specific coregulator complexes to the promoters of cell cycle genes. The coactivator-associated arginine methyltransferase CARM1 is a positive regulator of ER{alpha}-mediated transcriptional activation. Here, we show that CARM1 is essential for estrogen-induced cell cycle progression in the MCF-7 breast cancer cell line. CARM1 is specifically required for the estrogen-induced expression of the critical cell cycle transcriptional regulator E2F1 whereas estrogen stimulation of cyclin D1 is CARM1 independent. Upon estrogen stimulation, the E2F1 promoter is subject to CARM1-dependent dimethylation on histone H3 arginine 17 (H3R17me2) in a process that parallels the recruitment of ER{alpha}. Additionally, we find that the recruitment of CARM1 and subsequent histone arginine dimethylation are dependent on the presence of the oncogenic coactivator AIB1. Thus, CARM1 is a critical factor in the pathway of estrogen-stimulated breast cancer growth downstream of ER{alpha} and AIB1 and upstream of the cell cycle regulatory transcription factor E2F1. These studies identify CARM1 as a potential new target in the treatment of estrogen-dependent breast cancer. [Cancer Res 2008;68(1):301–6]


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Human breast cancer proliferation is a complex process involving pathways that are controlled by hormones, growth factors, and other signaling molecules (1). The steroid hormone 17β-estradiol (E2) regulates gene expression through the activation of the estrogen receptors {alpha} and β (ER{alpha} and ERβ), which function as ligand-regulated transcription factors (2). In breast cancer cells, E2-activated ER{alpha} modulates the expression of key cell cycle regulatory genes, including cyclin D1 (CCND1) and the transcription factor E2F1 (3–6). These factors play important roles in cell cycle progression, directing the phosphorylation and inactivation of the retinoblastoma protein (Rb) and mediating the expression of genes involved in DNA replication and S phase entry (7), respectively. Hence, drugs that block ER{alpha} action such as tamoxifen have been widely used in the prevention and treatment of breast cancer (8).

In the past few years, research interests have been directed toward understanding the roles of ER{alpha} (9, 10) and coregulatory proteins in breast cancer cell proliferation (11). Recently, the coactivator protein AIB1 (also called ACTR/SRC-3 and NCoA3), which belongs to the family of p160 steroid receptor coactivators, was shown to play an important role in breast cancer proliferation (12). AIB1 interacts with ligand-activated ER{alpha} and mediates the assembly of coactivator complexes containing chromatin-modifying enzymes, including the protein arginine methyltransferase PRMT4/CARM1 (13, 14). E2 stimulation induces the formation of a complex including CARM1, ER{alpha}, and AIB1 on the E2-responsive promoter of the TFF1 gene where CARM1 is capable of methylating specific arginines on histone H3 (15). Although CARM1 has a role in E2-mediated TFF1 gene activation, it is not known if CARM1 contributes to E2-induced breast cancer proliferation.

Here, we show that CARM1 is necessary for the E2-induced proliferation of breast cancer cells. We show that CARM1 is specifically required for the E2-mediated expression of E2F1 and not CCND1. Upon E2 signaling, the nucleosomes that are positioned near the transcription start site of the E2F1 gene undergo cell cycle variations in histone arginine dimethylation on arginine 17 of histone H3 (H3R17me2), which correlate with the recruitment of ER{alpha} and E2F1 and increased E2F1 expression. Collectively, our data suggest that CARM1 plays a critical role in breast cancer cell proliferation through the positive regulation of E2F1 expression, and thus may be an important new target in the treatment of estrogen-dependent breast cancer.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Reagents and antibodies. E2 and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma. The following antibodies were used: AIB1 (clone 34) was purchased from BD Biosciences; CARM1 (M-16) and CARM1 from Santa Cruz Biotechnology and Upstate Biotechnology, respectively; H3R17me2 from Upstate Biotechnology; and calnexin from Stressgen.

RNA interference. MCF-7 cells were transfected with small interfering RNA oligonucleotide duplexes (siRNA) at a final concentration of 50 nmol/L using LipofectAMINE 2000 (Invitrogen). Forty-eight hours after transfection, cells were stimulated and harvested for analysis. Cells were hormone deprived for at least 3 days at the moment of stimulation. To silence CARM1 expression, we used the ON-TARGETplus SMARTpool (Dharmacon; siCARM1-1) and Silencer siRNA (Ambion; siCARM1-2), sense 5'-GGAUAGAAAUCCCAUUCAAdTdT-3'. AIB1 was silenced as described in ref. 16.

Chromatin immunoprecipitation assays. Cells were hormone-deprived by culture for 3 to 4 days in phenol red–free medium (Invitrogen) supplemented with 5% charcoal-dextran–treated fetal bovine serum. Cells were stimulated with hormone for 45 min and cross-linked using 1% formaldehyde. Cells were lysed and sonicated three times for 10 s each at 12% amplitude (Fisher Sonic dismembrator, model 500). Immunoprecipitation, reverse cross-linking, and DNA purification were done as described (9). Immunoprecipitated DNA was quantified by real-time PCR.

Real-time PCR. Real-time PCR was done using SYBR Green PCR Master Mix (Applied Biosystems). The presence of a single amplicon was systemically verified by dissociation curve analysis. See Supplementary Table S1 for a list of chromatin immunoprecipitation primers used in this study.

Real-time reverse transcription-PCR. RNA isolation and real-time reverse transcription-PCR (RT-PCR) were done as described in ref. 17. Expression was normalized to RPS28 and glyceraldehyde-3-phosphate dehydrogenase. See Supplementary Table S1 for a list of target primers used in this study.

Western blotting. Western blot assays were done as described in ref. 17.

Cell cycle distribution and growth assays. MCF-7 cell cycle distribution was analyzed 24 h after stimulation with 10 nmol/L E2 or vehicle using propidium iodide staining and flow cytometry as described in ref. 17. For growth analysis, MCF-7 cells transfected with the indicated siRNAs were replated 48 h after transfection (seeding density, 3 x 104 cells per well) in phenol red–free DMEM containing 5% charcoal-stripped serum and 10 nmol/L E2. Cells were supplemented with fresh medium containing hormones every 48 h throughout the course of the experiment. The total cell number was quantified with MTT viability assay after 3 days of E2 stimulation and is expressed as percent change relative to control (siCONTROL-transfected cells treated with vehicle).

Data analysis. Data analysis was done using the Prism software. Statistical significance was determined using Student's t test comparison for unpaired data and was indicated as follows: *, P < 0.05; **, P < 0.01.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
CARM1 is required for E2-induced proliferation in ER{alpha}-positive breast cancer cells. CARM1 is thought to play an important role in hormone signaling through coactivation of nuclear receptor–mediated transcription and histone modification (18); however, this has been shown for a small number of target genes. Whereas both ER{alpha} and AIB1 have been shown to be involved in regulating cellular growth properties in both normal and breast cancer cells (19, 20), the mechanisms involving CARM1 in this process and its critical targets are not clearly defined. To explore the role of CARM1 in ER{alpha}-positive breast cancer cells, we examined the consequences of its suppression on the growth stimulatory effect of E2 using MCF-7 cells as a model. Expression of CARM1 was efficiently and specifically silenced in MCF-7 cells at the mRNA (data not shown) and protein levels (Fig. 1A ) with either of two different CARM1 siRNA oligonucleotide duplexes. Upon silencing of CARM1, we observed that the E2-mediated stimulation of MCF-7 cell cycle progression was strongly reduced (Fig. 1B). Furthermore, we observe a decrease in the number of MCF-7 cells following growth stimulation with E2 when CARM1 was silenced compared with a control siRNA (Fig. 1C). We found a similar effect of CARM1 silencing on E2-induced proliferation of T47D cells, another model of ER{alpha}-positive breast cancer (Supplementary Fig. S1). These data point to a critical role for CARM1 in the proliferative response to estrogen in ER{alpha}-positive breast cancer cells.


Figure 1
View larger version (19K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 1. Effect of CARM1 silencing in MCF-7 cells on E2-induced cell cycle progression. A, siRNA-mediated silencing of CARM1 in MCF-7 cells. Two different CARM1-specific siRNAs or a control siRNA were transfected into hormone-depleted MCF-7 cells. Extracts were collected and Western blotting was done with anti-CARM1 antibodies. Anti-calnexin was used as a loading control. B, cell cycle analysis of CARM1-depleted MCF-7 cells. MCF-7 cells were transfected as described in A. Forty-eight hours later, cells were treated with 10 nmol/L E2 or ethanol vehicle for 24 h and harvested to analyze DNA content by propidium iodide staining and flow cytometry. *, P < 0.05, versus siCONTROL-transfected cells treated with E2. C, cell viability assays were done as described in Materials and Methods using MCF-7 cells transfected with the indicated siRNAs. Cell proliferation is expressed as the percentage of the cells compared with the siCONTROL-transfected wells treated with E2. Columns, mean from one representative experiment done in triplicate; bars, SE. **, P < 0.01, versus similarly treated siCONTROL-transfected cells.

 
CARM1 controls breast cancer proliferation by regulating E2F1 expression. CARM1 has been shown to regulate the transcription of a small number of genes in response to distinct stimuli (21–24). To address whether CARM1 regulates the cell cycle by modulating transcription of specific genes that are involved in this process, we examined the effect of CARM1 depletion on the expression of specific cell cycle genes. Previous results show that the key cell cycle regulator E2F1 and, subsequently, its downstream target genes are critical for hormone regulation of the proliferative program of ER{alpha}-positive breast cancer cells (25). We therefore investigated the effect of CARM1 silencing on the expression of E2F1. As a positive control, CARM1 silencing reduced expression of the TFF1 gene (Fig. 2A ). Estrogen stimulation induced a 5- and 7-fold increase in E2F1 mRNA after 3 and 12 h, respectively, and this effect was very significantly reduced by CARM1 silencing (Fig. 2B). Interestingly, whereas CARM1 silencing impaired the transcription of E2F1, the estrogen-induced transcription of the CCND1 gene was unaffected by CARM1 silencing (Fig. 2C). Accordingly, we also observed that E2F1 protein expression levels were strongly reduced by CARM1 silencing (Fig. 2D). E2F1 expression was also diminished when CARM1 was silenced in another breast cancer cell model, the T47D cell line (Supplementary Fig. S2).


Figure 2
View larger version (21K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 2. CARM1 silencing affects E2-induced expression of E2F1. A to C, CARM1-specific or control siRNAs were transfected into hormone-depleted MCF-7 cells. Forty-eight hours after transfection, cells were treated with vehicle or 10 nmol/L E2 for the indicated time period, and RNA was isolated to measure the expression of TFF1 (A), E2F1 (B), and CCND1 (C) genes by real-time RT-PCR. Columns, mean of three independent replicates; bars, SD. Shown are results obtained from siCARM1-1 transfections; similar results were obtained from siCARM1-2 transfections. *, P < 0.05, versus siCONTROL-transfected cells. D, after 48 h of transfection with siCARM1 or control siRNA, MCF-7 cells were stimulated with 10 nmol/L E2 for 0, 24, and 48 h and whole-cell extracts from MCF-7 cells were analyzed by Western blot for E2F1.

 
CARM1 silencing blocks the E2 induction of E2F1-responsive genes. Estrogen regulation of E2F1 is important for the regulation of several cell cycle–associated genes. Having shown that CARM1 is important for the estrogen regulation of E2F1, we examined whether CARM1 silencing resulted in reduced expression of the E2F1-responsive genes including CCNE1, CCNE2, CDC25A, and CCNA1. As predicted in E2-stimulated MCF-7 cells in which CARM1 was silenced, we observed a significant reduction in the levels of CCNE1 and CCNE2 mRNA, as well as CDC25A and CCNA1 mRNA, compared with the control (Fig. 3 ). Hence, these data support an important role for CARM1 in the regulation of E2F1 and E2F1-responsive genes.


Figure 3
View larger version (21K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 3. Effect of CARM1 silencing on the E2-mediated expression of E2F1-responsive genes. siRNA-mediated silencing of CARM1 results in a reduced level of E2F1-responsive mRNAs. As in Fig. 2, CARM1 was silenced by siRNA transfection and the relative levels of CCNE1, CCNE2, CDC25A, and CCNA1 were determined by real-time RT-PCR. *, P < 0.05, versus siCONTROL-transfected cells.

 
CARM1 directly mediates the E2-induced activation of the E2F1 promoter. To examine the mechanism by which CARM1 is involved in the E2-mediated control of E2F1 transcription, we carried out chromatin immunoprecipitation assays. Previous studies have shown that a region up to –220 bp from the transcription start site is sufficient for full promoter activity (26, 27). This region contains two palindromic E2F-binding sites, several putative binding sites for SP1, a putative binding site for nuclear factor {kappa}B, and two canonical CAAT boxes. The E2F1 promoter has neither a TATA motif nor an initiator element. We find that E2 enhances the recruitment of both ER{alpha} and E2F1 to the E2F1 promoter (Fig. 4A ). In addition, we observe constitutive recruitment of SP1 to the E2F1 promoter independent of E2 stimulation (Fig. 4A). Because both AIB1 and CARM1 are thought to function in ER{alpha}- and E2F1-mediated transcriptional activation, we measured the enrichment of AIB1, CARM1, and H3R17me2 at the E2F1 promoter before and after E2 stimulation (Fig. 4B). Following E2 treatment, the coactivators AIB1 and CARM1, as well as the CARM1-catalyzed histone modification H3R17me2, are enriched at the E2F1 promoter (Fig. 4B). These results indicate that E2-mediated expression of E2F1 involves the recruitment of AIB1 and CARM1 to the E2F1 promoter and that this region of chromatin is subject to estrogen-dependent H3R17 dimethylation (H3R17me2). Interestingly, CARM1 was also recruited to the promoter of several E2F1 target genes (CCNE1, CCNE2, CDC25A, and CCNA1) following E2 stimulation (Supplementary Fig. S3). This suggests that in addition to the role of CARM1 in regulating E2F1 expression, it may play a second role in the regulation of E2F1 target genes. These results provide a direct link of CARM1 to cell cycle regulation through the transcriptional activation of E2F1 and E2F1 target genes.


Figure 4
View larger version (11K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 4. E2-mediated E2F1 expression corresponds with arginine methylation of nucleosomal histones located at the E2F1 promoter. A, chromatin immunoprecipitation analysis of the human E2F1 promoter. Chromatin immunoprecipitation was done with primers specific for the proximal portion of the E2F1 promoter with antibodies against Sp1, E2F1, and ER{alpha}. Formaldehyde cross-linked chromatin samples prepared from vehicle- and E2-treated MCF-7 cells were used for chromatin immunoprecipitation analyses with the indicated antibodies and analyzed by real-time PCR for the presence of the E2F1 promoter fragment as diagramed. Columns, mean of three independent replicates; bars, SD. B, chromatin immunoprecipitation analysis of the E2F1 promoter with antibodies against AIB1, CARM1, and H3R17me2.

 
CARM1 recruitment and E2-mediated histone modification at the E2F1 promoter are dependent on AIB1. It is thought that CARM1 is targeted to ER{alpha}-regulated genes via an interaction with SRC/p160 members including AIB1 (28). To test the requirement of AIB1 for CARM1 recruitment and histone arginine dimethylation at the E2F1 promoter, we silenced AIB1 in MCF-7 cells using AIB1-specific siRNAs and carried out CARM1 and H3R17me2 chromatin immunoprecipitation assays. Following efficient silencing of AIB1, we detect a significantly reduced recruitment of CARM1 to the E2F1 promoter compared with cells transfected with control siRNA (Fig. 5A ) both in the absence of hormone and, more significantly, after E2 stimulation. Importantly, the reduction of AIB1 had no measurable effect on the levels of CARM1 protein (Fig. 5A, right). Additionally, silencing AIB1 reduced the levels of H3R17me2 at the E2F1 promoter. The silencing of AIB1 affects histone methylation to a similar degree as when CARM1 is silenced (Fig. 5B). Thus, AIB1 functions in the recruitment of CARM1 and the subsequent H3R17 dimethylation during transcriptional activation of E2F1 and may in part explain the oncogenic functions of AIB1.


Figure 5
View larger version (21K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 5. Depletion of AIB1 inhibits CARM1 recruitment and E2F1 promoter methylation. A, siRNA-mediated depletion of AIB1 affects CARM1 recruitment to the E2F1 promoter. Specific siRNAs targeting AIB1 or control siRNAs were transfected into MCF-7 cells 48 h before E2 stimulation. CARM1 chromatin immunoprecipitation (ChIP) analyses for the E2F1 promoter were done as in Fig. 4. Western blot analysis of AIB1 and CARM1 was done with AIB1- and CARM1-specific antibodies. Calnexin was used as a loading control. *, P < 0.05, versus siCONTROL-transfected cells treated with E2. B, depletion of CARM1 or AIB1 results in decreased levels of H3R17me2 at the promoter of E2F1. siRNAs targeting a nonspecific control, CARM1, or AIB1 were transfected into MCF-7 cells and H3R17me2 chromatin immunoprecipitation was done. *, P < 0.05, versus siCONTROL-transfected cells treated with E2.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
Estrogens elicit proliferative responses in breast cancer cells through ER{alpha}-mediated transcriptional mechanisms (29). CARM1 has been implicated in the positive regulation of ER{alpha}-mediated gene activation in response to estrogen signaling (28). Thus, a presumed but not fully tested model for CARM1 action is that it functions in the control of cell proliferation through the activation of key cell cycle genes. In the current study, we describe a specific role for CARM1 in regulating the cell cycle by inducing the expression of E2F1 in MCF-7 cells following E2 treatment.

We show that CARM1 is required for the E2-mediated activation of E2F1 and for the induction of E2F1 target genes, including CDC25A, CCNA1, CCNE1, and CCNE2. These data are in agreement with a recent report showing that CARM1 functions in E2F1-mediated transactivation through AIB1 to positively regulate the expression of CCNE1 on cell cycle entry (22). Transfection of siRNA oligonucleotide duplexes into MCF-7 cells reduced the mRNA and protein levels of CARM1 by >70%. Reduced levels of CARM1 were sufficient to affect the E2-mediated activation of the TFF1 gene, suggesting that CARM1 was functionally silenced. Significantly, CARM1 silencing does not affect the E2-stimulation of CCND1, showing that CARM1 does not function in the entire E2 response but rather has an important role in targeting a subset of ER{alpha} targets including E2F1. The observed gene-specific effect of CARM1 is consistent with the reported promoter-specific role for CARM1 in the activation of specific nuclear factor {kappa}B–dependent genes (23).

At the E2-responsive TFF1 promoter, CARM1 recruitment was shown to follow ER{alpha} recruitment. Thus, CARM1 is thought to be recruited by ER{alpha} to this promoter (15, 30). However, it is not known whether the recruitment of CARM1 occurs at additional E2-responsive sites including nonclassical ER{alpha} target sites. Here, we show that CARM1 is recruited to the E2F1 promoter, a nonclassical ER{alpha} site (6). Our chromatin immunoprecipitation data confirm the recruitment of ER{alpha} to the promoter of E2F1 and show that E2 induces the recruitment of both AIB1 and CARM1. CARM1 targeting to the E2F1 promoter coincides with enhanced levels of H3R17 dimethylation. Importantly, H3R17me2 at this site is undetectable in CARM1-depleted cells, signifying that CARM1 is the main or sole enzyme responsible for the dimethylation of histone H3 at arginine 17. We also observe a major defect in CARM1 recruitment to the E2F1 promoter in AIB1-knockdown cells and that histone dimethylation at H3R17 is eliminated in AIB1-silenced cells on E2 stimulation. These data are in agreement with a model in which p160 coactivators such as AIB1 serve as scaffold factors to target CARM1 to specific regulated sites where CARM1 can methylate histones and participate in gene activation.

Methylation of histones by PRMTs is increasingly being acknowledged as an important aspect for the dynamic regulation of gene expression. For example, CARM1/PRMT4 and PRMT1, two type I PRMTs, have been proposed to play a role in gene activation and were shown to catalyze monomethylation and asymmetrical NG,NG-arginine dimethylation on histone H3R2, H3R17, H3R26, and H4R3, respectively (31). In contrast, PRMT5, a type II PRMT, was shown to catalyze histone H4 monomethylation and symmetrical NG,N1G-arginine dimethylation, a modification that coincides with gene repression (32, 33). Whereas it is recognized that specific PRMT enzymes are important transcriptional coregulators, it is unclear how these enzymes and their associated histone modifications function to coordinately regulate gene expression in vivo. In this report, we reveal that the arginine methyltransferase CARM1 functions in breast cancer proliferation through specific gene activation.

Aberrant expression of CARM1 has been linked to human prostate and breast cancers (21, 22, 34). Consistent with a study showing that CARM1–/– murine embryonic fibroblasts display an altered capacity to enter the cell cycle upon serum stimulation (22), we show that CARM1 depletion also adversely affects breast cancer cell proliferation when stimulated with estrogen. Our finding that CARM1 is required for E2F1 expression, but not CCND1 expression, following estrogen simulation suggests that the pathway of estrogen-stimulated growth involves the direct transcriptional effect of ER{alpha}, AIB1, and CARM1 on the E2F1 promoter. In this model (Fig. 6 ), the ability of estrogen acting through ER{alpha} and AIB1 to activate CCND1 (35) and subsequently inactivate Rb is independent of CARM1, although the ability of estrogen to stimulate the cell cycle is dependent on CARM1 activation of E2F1 and several of its downstream targets. Taken together, these results suggest that CARM1 may represent an important target for therapeutic intervention in hormone-dependent breast cancer.


Figure 6
View larger version (7K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 6. Model of E2-mediated breast cancer cell proliferation.

 

   Acknowledgments
 
Grant support: National Institutes of Diabetes, Digestive, and Kidney Diseases grant R56DK074967 (M. Brown); NIH (P.A. Silver); National Cancer Institute (Dana-Farber/Harvard Cancer Center Breast Cancer Specialized Program of Research Excellence Grant), National Cancer Institute grant P01CA8011105 (M. Brown), Dana-Farber Cancer Institute Women's Cancers Program, and National Science Foundation predoctoral fellowship (S. Frietze).

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

We thank I. Swinburne and J. Eeckhoute for critical reading of the manuscript and the members of the Brown and Silver laboratory for helpful discussions.


   Footnotes
 
Note: Supplementary data for this article are available at Cancer Research Online (http://cancerres.aacrjournals.org/).

Received 6/ 5/07. Revised 10/ 2/07. Accepted 11/ 1/07.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Doisneau-Sixou SF, Sergio CM, Carroll JS, Hui R, Musgrove EA, Sutherland RL. Estrogen and antiestrogen regulation of cell cycle progression in breast cancer cells. Endocr Relat Cancer 2003;10:179–86.[Abstract]
  2. Klinge CM. Estrogen receptor interaction with estrogen response elements. Nucleic Acids Res 2001;29:2905–19.[Abstract/Free Full Text]
  3. Altucci L, Addeo R, Cicatiello L, et al. 17β-Estradiol induces cyclin D1 gene transcription, p36D1-34cdk4 complex activation and p105Rb phosphorylation during mitogenic stimulation of G(1)-arrested human breast cancer cells. Oncogene 1996;12:2315–24.[Medline]
  4. Eeckhoute J, Carroll JS, Geistlinger TR, Torres-Arzayus MI, Brown M. A cell-type-specific transcriptional network required for estrogen regulation of cyclin D1 and cell cycle progression in breast cancer. Genes Dev 2006;20:2513–26.[Abstract/Free Full Text]
  5. Musgrove EA, Hamilton JA, Lee CS, Sweeney KJ, Watts CK, Sutherland RL. Growth factor, steroid, and steroid antagonist regulation of cyclin gene expression associated with changes in T-47D human breast cancer cell cycle progression. Mol Cell Biol 1993;13:3577–87.[Abstract/Free Full Text]
  6. Wang W, Dong L, Saville B, Safe S. Transcriptional activation of E2F1 gene expression by 17β-estradiol in MCF-7 cells is regulated by NF-Y-Sp1/estrogen receptor interactions. Mol Endocrinol 1999;13:1373–87.[Abstract/Free Full Text]
  7. Dimova DK, Dyson NJ. The E2F transcriptional network: old acquaintances with new faces. Oncogene 2005;24:2810–26.[CrossRef][Medline]
  8. Wickerham L. Tamoxifen—an update on current data and where it can now be used. Breast Cancer Res Treat 2002;75 Suppl 1:S7–12; discussion S33–5.[CrossRef][Medline]
  9. Carroll JS, Liu XS, Brodsky AS, et al. Chromosome-wide mapping of estrogen receptor binding reveals long-range regulation requiring the forkhead protein FoxA1. Cell 2005;122:33–43.[CrossRef][Medline]
  10. Carroll JS, Meyer CA, Song J, et al. Genome-wide analysis of estrogen receptor binding sites. Nat Genet 2006;38:1289–97.[CrossRef][Medline]
  11. Green KA, Carroll JS. Oestrogen-receptor-mediated transcription and the influence of co-factors and chromatin state. Nat Rev 2007;7:713–22.[CrossRef]
  12. Louie MC, Zou JX, Rabinovich A, Chen HW. ACTR/AIB1 functions as an E2F1 coactivator to promote breast cancer cell proliferation and antiestrogen resistance. Mol Cell Biol 2004;24:5157–71.[Abstract/Free Full Text]
  13. Shang Y, Hu X, DiRenzo J, Lazar MA, Brown M. Cofactor dynamics and sufficiency in estrogen receptor-regulated transcription. Cell 2000;103:843–52.[CrossRef][Medline]
  14. Teyssier C, Chen D, Stallcup MR. Requirement for multiple domains of the protein arginine methyltransferase CARM1 in its transcriptional coactivator function. J Biol Chem 2002;277:46066–72.[Abstract/Free Full Text]
  15. Metivier R, Penot G, Hubner MR, et al. Estrogen receptor-{alpha} directs ordered, cyclical, and combinatorial recruitment of cofactors on a natural target promoter. Cell 2003;115:751–63.[CrossRef][Medline]
  16. Shao W, Keeton EK, McDonnell DP, Brown M. Coactivator AIB1 links estrogen receptor transcriptional activity and stability. Proc Natl Acad Sci U S A 2004;101:11599–604.[Abstract/Free Full Text]
  17. Keeton EK, Brown M. Cell cycle progression stimulated by tamoxifen-bound estrogen receptor-{alpha} and promoter-specific effects in breast cancer cells deficient in N-CoR and SMRT. Mol Endocrinol 2005;19:1543–54.[Abstract/Free Full Text]
  18. Stallcup MR. Role of protein methylation in chromatin remodeling and transcriptional regulation. Oncogene 2001;20:3014–20.[CrossRef][Medline]
  19. Katzenellenbogen BS, Montano MM, Ekena K, Herman ME, McInerney EM, William L. McGuire Memorial Lecture. Antiestrogens: mechanisms of action and resistance in breast cancer. Breast Cancer Res Treat 1997;44:23–38.[Medline]
  20. Yan J, Tsai SY, Tsai MJ. SRC-3/AIB1: transcriptional coactivator in oncogenesis. Acta Pharmacol Sin 2006;27:387–94.[CrossRef][Medline]
  21. Majumder S, Liu Y, Ford OH III, Mohler JL, Whang YE. Involvement of arginine methyltransferase CARM1 in androgen receptor function and prostate cancer cell viability. Prostate 2006;66:1292–301.[CrossRef][Medline]
  22. El Messaoudi S, Fabbrizio E, Rodriguez C, et al. Coactivator-associated arginine methyltransferase 1 (CARM1) is a positive regulator of the cyclin E1 gene. Proc Natl Acad Sci U S A 2006;103:13351–6.[Abstract/Free Full Text]
  23. Covic M, Hassa PO, Saccani S, et al. Arginine methyltransferase CARM1 is a promoter-specific regulator of NF-{kappa}B-dependent gene expression. EMBO J 2005;24:85–96.[CrossRef][Medline]
  24. Ma H, Baumann CT, Li H, et al. Hormone-dependent, CARM1-directed, arginine-specific methylation of histone H3 on a steroid-regulated promoter. Curr Biol 2001;11:1981–5.[CrossRef][Medline]
  25. Stender JD, Frasor J, Komm B, Chang KC, Kraus WL, Katzenellenbogen BS. Estrogen-regulated gene networks in human breast cancer cells: involvement of E2F1 in the regulation of cell proliferation. Mol Endocrinol 2007;21:2112–23.[Abstract/Free Full Text]
  26. Neuman E, Flemington EK, Sellers WR, Kaelin WG, Jr. Transcription of the E2F-1 gene is rendered cell cycle dependent by E2F DNA-binding sites within its promoter. Mol Cell Biol 1994;14:6607–15.[Abstract/Free Full Text]
  27. Johnson DG, Ohtani K, Nevins JR. Autoregulatory control of E2F1 expression in response to positive and negative regulators of cell cycle progression. Genes Dev 1994;8:1514–25.[Abstract/Free Full Text]
  28. Stallcup MR, Kim JH, Teyssier C, Lee YH, Ma H, Chen D. The roles of protein-protein interactions and protein methylation in transcriptional activation by nuclear receptors and their coactivators. J Steroid Biochem Mol Biol 2003;85:139–45.[CrossRef][Medline]
  29. Katzenellenbogen BS, Montano MM, Ediger TR, et al. Estrogen receptors: selective ligands, partners, and distinctive pharmacology. Recent Prog Horm Res 2000;55:163–93; discussion 94–5.[Medline]
  30. Daujat S, Bauer UM, Shah V, Turner B, Berger S, Kouzarides T. Crosstalk between CARM1 methylation and CBP acetylation on histone H3. Curr Biol 2002;12:2090–7.[CrossRef][Medline]
  31. Bedford MT, Richard S. Arginine methylation an emerging regulator of protein function. Mol Cell 2005;18:263–72.[CrossRef][Medline]
  32. Fabbrizio E, El Messaoudi S, Polanowska J, et al. Negative regulation of transcription by the type II arginine methyltransferase PRMT5. EMBO Rep 2002;3:641–5.[CrossRef][Medline]
  33. Pal S, Vishwanath SN, Erdjument-Bromage H, Tempst P, Sif S. Human SWI/SNF-associated PRMT5 methylates histone H3 arginine 8 and negatively regulates expression of ST7 and NM23 tumor suppressor genes. Mol Cell Biol 2004;24:9630–45.[Abstract/Free Full Text]
  34. Hong H, Kao C, Jeng MH, et al. Aberrant expression of CARM1, a transcriptional coactivator of androgen receptor, in the development of prostate carcinoma and androgen-independent status. Cancer 2004;101:83–9.[CrossRef][Medline]
  35. Planas-Silva MD, Shang Y, Donaher JL, Brown M, Weinberg RA. AIB1 enhances estrogen-dependent induction of cyclin D1 expression. Cancer Res 2001;61:3858–62.[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 Mol. Endocrinol.Home page
S. C. Wu and Y. Zhang
Minireview: Role of Protein Methylation and Demethylation in Nuclear Hormone Signaling
Mol. Endocrinol., September 1, 2009; 23(9): 1323 - 1334.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
A. Scoumanne, J. Zhang, and X. Chen
PRMT5 is required for cell-cycle progression and p53 tumor suppressor function
Nucleic Acids Res., August 1, 2009; 37(15): 4965 - 4976.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
M. Lupien, J. Eeckhoute, C. A. Meyer, S. A. Krum, D. R. Rhodes, X. S. Liu, and M. Brown
Coactivator Function Defines the Active Estrogen Receptor Alpha Cistrome
Mol. Cell. Biol., June 15, 2009; 29(12): 3413 - 3423.
[Abstract] [Full Text] [PDF]

Home page
 Endocr Relat CancerHome page
M. Lupien and M. Brown
Cistromics of hormone-dependent cancer
Endocr. Relat. Cancer, June 1, 2009; 16(2): 381 - 389.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
Z. Yu, T. Chen, J. Hebert, E. Li, and S. Richard
A Mouse PRMT1 Null Allele Defines an Essential Role for Arginine Methylation in Genome Maintenance and Cell Proliferation
Mol. Cell. Biol., June 1, 2009; 29(11): 2982 - 2996.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
Y.-H. Lee and M. R. Stallcup
Minireview: Protein Arginine Methylation of Nonhistone Proteins in Transcriptional Regulation
Mol. Endocrinol., April 1, 2009; 23(4): 425 - 433.
[Abstract] [Full Text] [PDF]

Home page
 Genome ResHome page
J. Eeckhoute, M. Lupien, C. A. Meyer, M. P. Verzi, R. A. Shivdasani, X. S. Liu, and M. Brown
Cell-type selective chromatin remodeling defines the active subset of FOXA1-bound enhancers
Genome Res., March 1, 2009; 19(3): 372 - 380.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
C. S. Chan and J. S. Song
CCCTC-Binding Factor Confines the Distal Action of Estrogen Receptor
Cancer Res., November 1, 2008; 68(21): 9041 - 9049.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
A. K. L. Cheung, H. L. Lung, S. C. Hung, E. W. L. Law, Y. Cheng, W. L. Yau, D. K. Bangarusamy, L. D. Miller, E. T.-B. Liu, J.-Y. Shao, et al.
Functional Analysis of a Cell Cycle-Associated, Tumor-Suppressive Gene, Protein Tyrosine Phosphatase Receptor Type G, in Nasopharyngeal Carcinoma
Cancer Res., October 1, 2008; 68(19): 8137 - 8145.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Supplementary Data
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Frietze, S.
Right arrow Articles by Brown, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Frietze, S.
Right arrow Articles by Brown, M.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online