The Journal of Clinical Investigation http://www.jci.org Volume 122 Number 8 August 2012
PES1 promotes breast cancer by differentially
regulating ERα and ERβ
Long Cheng,1,2 Jieping Li,1,3 Yongjian Han,1 Jing Lin,4 Chang Niu,1 Zhichao Zhou,1 Bin Yuan,1
Ke Huang,5 Jiezhi Li,1 Kai Jiang,1 Hao Zhang,1 Lihua Ding,1 Xiaojie Xu,1 and Qinong Ye1,2
1Department of Medical Molecular Biology, Beijing Institute of Biotechnology, Beijing, People’s Republic of China. 2Institute of Cancer Stem Cell, Cancer Center,
Dalian Medical University, Liaoning, People’s Republic of China. 3Department of Clinical Laboratory, Fuzhou General Hospital of Fujian Corps,
CAPF, Fuzhou, People’s Republic of China. 4Department of Clinical Laboratory, First Affiliated Hospital, and 5Department of Obstetrics and Gynecology,
Chinese PLA General Hospital, Beijing, People’s Republic of China.
The initiation of breast cancer is associated with increased expression of tumor-promoting estrogen receptor α
(ERα) protein and decreased expression of tumor-suppressive ERβ protein. However, the mechanism underly-
ing this process is unknown. Here we show that PES1 (also known as Pescadillo), an estrogen-inducible protein
that is overexpressed in breast cancer, can regulate the balance between ERα and ERβ. We found that PES1
modulated many estrogen-responsive genes by enhancing the transcriptional activity of ERα while inhibiting
transcriptional activity of ERβ. Consistent with this regulation of ERα and ERβ transcriptional activity, PES1
increased the stability of the ERα protein and decreased that of ERβ through the ubiquitin-proteasome path-
way, mediated by the carboxyl terminus of Hsc70-interacting protein (CHIP). Moreover, PES1 transformed
normal human mammary epithelial cells and was required for estrogen-induced breast tumor growth in nude
mice. Further analysis of clinical samples showed that expression of PES1 correlated positively with ERα
expression and negatively with ERβ expression and predicted good clinical outcome in breast cancer. Our data
demonstrate that PES1 contributes to breast tumor growth through regulating the balance between ERα and
ERβ and may be a better target for the development of drugs that selectively regulate ERα and ERβ activities.
The association between estrogen and breast cancer was recognized
over 100 years ago. Estrogen exerts its function through its 2 nucle-
ar receptors, estrogen receptor α (ERα) and ERβ (1, 2). ER belongs
to a superfamily of ligand-activated transcription factors that share
structural similarity characterized by several functional domains.
N-terminal estrogen-independent and C-terminal estrogen-depen-
dent activation function domains (AF1 and AF2, respectively) con-
tribute to the transcriptional activity of the 2 receptors. The DNA-
binding domain of the ERs is centrally located. The ligand-binding
domain, overlapping AF2, shows 58% homology between ERα and
ERβ. The DNA-binding domain is identical between the 2 recep-
tors, except for 3 amino acids. However, the AF1 domain of ERβ
has only 28% homology with that of ERα. The binding of estrogen
to ER leads to ER dimerization and its recruitment to the estrogen-
responsive elements (EREs) on the promoters of ER target genes,
thereby either enhancing or repressing gene activation.
The development of breast cancer is associated with dysregu-
lation of ER expression (3–8). Compared with that in normal
breast tissues, the proportion of cells expressing ERα is increased,
whereas ERβ expression is reduced, in hormone-dependent breast
tumors. The ratio of ERα/ERβ expression is higher in breast
tumors than in normal tissues, and ERα and ERβ are antagonis-
tic to each other. ERα mediates the tumor-promoting effects of
estrogens, whereas ERβ inhibits breast cancer cell growth. ERβ
reduces cell proliferation induced by ERα activation. Although
ERα and ERβ have been shown to have a yin-yang relationship in
breast tumorigenesis, the molecular mechanism underlying this
process remains unclear.
In this study, we show that PES1 (also known as Pescadillo)
plays an essential role in estrogen-induced breast tumor growth
through regulation of the yin-yang balance between ERα and ERβ
and is the first such gene to be identified to our knowledge. PES1,
a breast cancer–associated gene 1 (BRCA1) C-terminal (BRCT)
domain-containing protein, is estrogen inducible, and its expres-
sion gradually increases during breast cancer development and
progression (9–11). Theoretically, in the treatment of patients
with ERα-positive breast cancer, in which ERβ is antagonistic
to ERα, a drug that decreases transcriptional activity of ERα but
increases that of ERβ should be better than the currently used
endocrine drugs tamoxifen or fulvestrant, which decrease both
ERα and ERβ transactivation (12, 13). We show that, through
the ubiquitin-proteasome pathway, PES1 enhances ERα levels
but reduces ERβ protein levels, correlating with their respective
physiological activities in breast cancer. Thus, PES1 may repre-
sent a very promising target for the development of better drugs
for breast cancer endocrine therapy.
PES1 differentially regulates transcriptional activity of ERα and ERβ as
well as their target genes. To define the exact role of PES1 in breast
tumor growth, we investigated whether PES1 regulates estrogen
signaling. PES1 overexpression in ERα- and ERβ-positive MCF7
cells (Figure 1A), ERα-positive and ERβ-negative ZR75-1 and
T47D cells (Supplemental Figure 1, A and B; supplemental mate-
rial available online with this article; doi:10.1172/JCI62676DS1),
and ERα- and ERβ-negative SKBR3 (Figure 1B) breast cancer cells
increased transcription of a luciferase reporter construct con-
taining the ERE in response to the ERα-specific agonist propyl-
pyrazole triol (PPT) but decreased ERE reporter transcription in
response to the ERβ-specific agonist diarylpropionitrile (DPN).
This effect was PES1 specific because expression of the known
Authorship note: Long Cheng and Jieping Li contributed equally to this work.
Conflict of interest: The authors have declared that no conflict of interest exists.
Citation for this article: J Clin Invest. 2012;122(8):2857–2870. doi:10.1172/JCI62676.
Related Commentary, page 2771
2858 The Journal of Clinical Investigation http://www.jci.org Volume 122 Number 8 August 2012
PES1 differentially regulates transcriptional activity of ERα and ERβ and expression of
their target genes. (A and B) Luciferase reporter assays of ERα and ERβ transcriptional
activity in (A) MCF7 or (B) SKBR3 cells transiently transfected with ERE-LUC and PES1,
SRC1, GRIP1, XRCC1, or BARD1 with or without ERα or ERβ and 24-hour treatment
with 10 nM E2, 1 nM PPT, or 1 nM DPN. Results shown are mean ± SD of 3 independent
experiments. ‡P < 0.01, *P < 0.01, #P < 0.01, †P < 0.01 versus empty vector in the (A)
absence or (B) presence of ERα or ERβ with vehicle (–), E2, PPT, and DPN, respectively.
(C) Luciferase reporter assays in MCF7 cells stably transfected with PES1 siRNA or PES1
siRNA plus siRNA-resistant PES1 (PES1-R) and treated as above. Immunoblot analysis of
PES1 expression is shown. Results shown are mean ± SD of 3 independent experiments.
*P < 0.01, #P < 0.01, †P < 0.01 versus control siRNA with E2, PPT, and DPN, respectively.
(D) Real-time RT-PCR analysis of 47 genes identified by cDNA microarray in our study and
4 genes identified in other studies (CCND1, CTSD, E2F1, and C-FOS) in PES1 knockdown
MCF7 cells treated or not treated with E2 (+E2 or –E2, respectively) for 24 hours. Data
shown are mean ± SD of triplicate measurements that have been repeated 3 times with
similar results. *P < 0.05, #P < 0.01 versus control siRNA without E2. ‡P < 0.05, †P < 0.01
versus control siRNA with E2. (E) Immunoblot analysis of estrogen-responsive gene expres-
sion in PES1 knockdown MCF7 cells.
The Journal of Clinical Investigation http://www.jci.org Volume 122 Number 8 August 2012
Program (2011CB504202 and 2012CB945100), National Natu-
ral Science Foundation (81072173, 30625035, and 30530320),
and National Key Technologies R&D Program for New Drugs
Received for publication December 30, 2011, and accepted in
revised form May 31, 2012.
Address correspondence to: Qinong Ye, Department of Medical
Molecular Biology, Beijing Institute of Biotechnology, 27 Tai-
Ping Lu Rd., Beijing 100850, China. Phone: 8610.6818.0809; Fax:
8610.6824.8045; E-mail: email@example.com.
vival curves were determined with the log-rank test. Statistical calculations
were performed using SPSS 13.0. P values of less than 0.05 were considered
Study approval. Animal studies were approved by the Institutional Animal
Care Committee of Beijing Institute of Biotechnology. Breast cancer sam-
ples were obtained from Chinese PLA General Hospital, with the informed
consent of patients and with institutional approval for experiments from
Chinese PLA General Hospital and Beijing Institute of Biotechnology.
We thank Rong Li and Haian Fu for helpful discussions. This
work was supported by Major State Basic Research Development
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