Biochemical and Biophysical Research Communications 469 (2016) 679e685

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Biochemical and Biophysical Research Communications

journal homepage: www.elsevier.com/locate/ybbrc

Inhibition of BRD4 suppresses tumor growth and enhances iodine

uptake in thyroid cancer
Xuemei Gao a, b, c, 1, Xinchao Wu c, 1, Xiao Zhang a, b, Wenjuan Hua a, b, Yajing Zhang a, b,
Yusufu Maimaiti d, Zairong Gao a, b, *, Yongxue Zhang a, b
a
Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province,
China
b
Hubei Province Key Laboratory of Molecular Imaging, Wuhan 430022, Hubei Province, China
c
Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
d
Department of Thyroid and Breast Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei
Province, China

a r t i c l e i n f o a b s t r a c t

Article history: Thyroid cancer is a common malignancy of the endocrine system. Although radioiodine 131I treatment on
Received 25 November 2015 differentiated thyroid cancer is widely used, many patients still fail to benefit from 131I therapy. There-
Accepted 1 December 2015 fore, exploration of novel targeted therapies to suppress tumor growth and improve radioiodine uptake
Available online 18 December 2015
remains necessary. Bromodomain-containing protein 4 (BRD4) is an important member of the bromo-
domain and extra terminal domain family that influences transcription of downstream genes by binding
Keywords:
to acetylated histones. In the present study, we found that BRD4 was up-regulated in thyroid cancer
BRD4
tissues and cell lines. Inhibition of BRD4 in thyroid cancer cells by JQ1 resulted in cell cycle arrest at G0/
JQ1
Thyroid cancer
G1 phase and enhanced 131I uptake in vitro and suppressed tumor growth in vivo. Moreover, JQ1 treat-
NIS ment suppressed C-MYC but enhanced NIS expression. We further demonstrated that BRD4 was enriched
C-MYC in the promoter region of C-MYC, which could be markedly blocked by JQ1 treatment. In conclusion, our
findings revealed that the aberrant expression of BRD4 in thyroid cancer is possibly involved in tumor
progression, and JQ1 is potentially an effective chemotherapeutic agent against human thyroid cancer.
© 2015 Elsevier Inc. All rights reserved.

1. Introduction uptake, and their conditions worsen dramatically [5]. Therefore,

Thyroid cancer (TC) is the most common malignancy of the
endocrine system, and the incidence of TC has increased steadily for
many years [1]. In America, there has been a threefold increase over
the last four decades [2]. To date, the contribution of gene dysre-
gulation in TC pathogenesis and tumor progression remains
incompletely understood [3]_ENREF_2. Current treatment strate-
gies for differentiated thyroid cancer (DTC) involve surgery,
thyrotropin suppressive therapy, and radioactive iodine (RAI) [4,5]
_ENREF_4. Despite the overall favorable prognosis, a considerable
part of DTC patients fail to benefit from RAI therapy owing to
consecutive tumor progression and decreased ability of radioiodine
* Corresponding author. Address: No. 1277, Jiefang Road, Wuhan 430022, Hubei
Province, China.
E-mail address: gaobonn@163.com (Z. Gao).
1
These authors contributed equally to this work.
exploration of new gene deregulations and new targeted therapies
remain necessary to enhance the survival of patients with DTC.
Bromodomain-containing protein 4 (BRD4) is a member of the
bromodomain and extra terminal domain (BET) family that in-
fluences transcription of downstream genes by binding to acety-
lated histones, which plays an important role in genesis and
development of many diseases including cancer. Deregulation of
BRD4 was frequently reported to influence cancer cell biological
activity and inhibition of BRD4 could effectively delay tumor
growth [6e8]. Currently, JQ1 and I-BET762 are two well-studied
synthetic small molecular anticancer drugs by binding to and
inhibiting the acetyl-lysine recognition site of BRD4 [9]. JQ1 is a
first-in-class inhibitor of BRD4, which engages to recruit tran-
scription elongation factor b (P-TEFb) complex to acetylated chro-
matin region and promotes transcription by interacting with RNA
polymerase II [10,11]. The effect of BRD4 inhibitor, resulting in cell
cycle arrest and/or apoptosis, has been clarified in several diseases,
such as leukemia, osteosarcoma, prostatic cancer and lung cancer

http://dx.doi.org/10.1016/j.bbrc.2015.12.008
0006-291X/© 2015 Elsevier Inc. All rights reserved.
680 X. Gao et al. / Biochemical and Biophysical Research Communications 469 (2016) 679e685
[12e15]_ENREF_8_ENREF_8. However, expression level and bio-
logical function of BRD4 in DTC are still largely unknown, and the
effect of BET inhibitor in treatment of DTC has yet to be reported.
In our study, we evaluated the expression levels of BRD4 in DTC
and assessed the therapeutic effect of pharmacologically sup-
pressing BRD4 protein in DTC in vitro and in vivo. Our original
findings suggest the significant effect of BRD4 inhibitor in DTC
treatment by influencing cellular viability and regulating the
expression of important cell-cycle and survival regulators. Shortly,
our results reveal a potential role for BRD4 in DTC biological activity
and establish a promising approach in therapeutic intervention
against this disease.
2. Materials and methods
2.1. Cell culture and human tissue specimens
The human Nthy Ori3-1 and PTC cell lines, BCPAP and K1, were
purchased from American Type Culture Collection (ATCC, Mana-
ssas, VA, USA). The cells were maintained in DMEM/F-12 (Gibco,
Grand Island, NY, US) supplemented with 10% fetal bovine serum,
penicillin G (100 U/mL), and streptomycin (100 mg/mL). Forty-five
pairs of human PTC lesions and surrounding normal thyroid tis-
sue specimens were obtained from PTC patients who underwent
total/near-total thyroidectomy at Department of Thyroid and Brest
Surgery of our hospital from 2013 to 2014. Histopathological re-
ports were assessed by at least two senior pathologists. Written
informed consent was obtained from the patients so that their
specimens could be further studied, and the research was approved
by the Ethics Committee of Union Hospital, Tongji Medical College,
Huazhong University of Science and Technology ([2013] IEC (376)).
2.2. Reagents and chemicals
JQ1, bromodomain and extraterminal (BET) inhibitor, was ob-
tained from Selleckchem (Catalog NO.S7110, USA), and diluted in
DMSO to a stock concentration of 10 mM at 4
C. Carrier-free
131
IeNaI was provided by Atomic hi-tech co., Ltd., Beijing, China.
2.3. Real-time PCR analysis of mRNA expression
Tissue specimens and cell line samples were stored frozen
at 80
C until use. Total RNA was extracted from specimens and
cell lines with TRIzol reagent (Invitrogen). Concentration and purity
of RNA were measured using Biophotometer (Eppendorf, Hamburg,
Germany). For analysis of BRD4 and NIS mRNA, reverse transcrip-
tion kit PrimeScript™ RT reagent Kit with gDNA Eraser (Takara
Biomedical Technology, Dalian, China) was used to synthesize
complementary DNA. Then the SYBR® Premix Ex Taq™ (Tli RNaseH
Plus) kit (Takara Biomedical Technology) was used for RT-PCR ap-
plications. Genes were amplified using the primers as follows:
BRD4: 50 -GTGGGAGGAAAGAAACAGGGACA-30 (forward) and 50 -
AGGAGGAGGATTCGGCTGAGG-30 (reverse); NIS: 50 -
GACAAACCTCTGAGGACAGGG-30 (forward) and 50 -ATACTGGG-
GACGGTTGAAGC-30 (reverse); C-MYC: 50 -AGGGATCGCGCTGAGTA-
TAA-30 (forward) and 50 -TGCCTCTCGCTGGAATTACT-30 (reverse).
GAPDH: 50 -TCAAGAAGGTGGTGAAGCAG-30 (forward) and 50 -
CGTCAAAGGTGGAGGAGTG-30 (reverse). The primers was synthe-
sized and purchased from Takara Biomedical Technology (Takara
Biomedical Technology, Dalian, China). All analyses were per-
formed by the StepOnePlus Real-Time PCR System (Applied Bio-
systems, Foster City, CA, USA). Results were normalized to GAPDH
and calculated as fold change using the DDCT method compared to
control cells.
2.4. Western blotting analysis
RIPA (Thermo Scientific, Rockford, IL, US) buffer was used to
extract total protein from tissues and cell lines. The BCA Protein
assay kit (Beyotime) was applied to determine protein concentra-
tion of each sample. Equal amounts of lysate protein sample was
subjected to each lane, electrophoresed in 10% SDS-PAGE, and
transferred onto PVDF membranes. And then the membranes were
incubated with primary antibodies (1:1000 dilution) over night at
4
C, washed (  3) with Tris-buffered saline with 0.1% Tween-20
(TBST), and incubated with HRP conjugated secondary antibodies
(1:400 dilution) at 37
C for 1 h, detected using the ECL kit (Beyo-
time). ImageJ software version 1.48 (NIH) was utilized to analysis
the expression levels of different proteins. The antibodies of BRD4
(ab128874), NIS (ab17795),C-MYC(ab39688) and P21(ab7960) were
purchased from Abcam (Cambridge, MA, USA), GAPDH (D16H11)
was purchased from Cell Signaling Technology Inc. (Vebery, MA,
USA). Secondary antibodies, anti-rabbit and anti-mouse IgG con-
jugated with horseradish peroxidase, were obtained from Wuhan
Boster Bio-engineering Limited Company, China.
2.5. Cell viability evaluation by MTT assay
Cell viability after JQ1 treatment was evaluated by the MTT
assay. Cells were seeded in 96-well plates at a density of
5  103 cells/0.1 mL per well. After drug treatment, each well was
added 20 mL MTT stock solutions. After incubation in the dark at
37
C for an additional 4 h, the supernatant was discarded and
100 mL DMSO was added to dissolve the precipitate. The absorbance
of the generated formazan OD at 570 nm was used to determine cell
viability.
2.6. Flow cytometry analysis of the cell cycle
Cells were treated with 0.25 mM, 0.5 mM and 1 mM JQ1. After 72 h
treatment, cells for cell cycle analysis were harvested and stained
with propidium iodine (PI, Sigma, St Louis, MO, USA) according to
the manufacturer's protocol. The ModFit LT software was utilized
for data analysis.
2.7. Iodine uptake assay
Radioiodine uptake was assayed as previously introduced.
Briefly, cells were seeded in 24-well plates (105 cells in 0.5 mL
medium) and treated with JQ1. After drug treatment, radioiodine
uptake studies were carried out. The cells were washed twice with
0.5 mL PBS, and then incubated with 0.5 mL serum-free DMEM/F-
12 containing 0.1 mCi carrier-free Na131I, with or without 10 mM
NaClO4. NaClO4 was used as an NIS-competitive inhibitor for radi-
oiodine uptake. After 1 h incubation at 37
C, the medium con-
taining radioiodine was removed and the cells were rinsed twice
with 1 mL PBS. Then, the cells in each well were lysed with NaOH,
washed twice with PBS, and the cell-associated radioactivity of the
collected lysed cells was measured using a gamma counter.
2.8. Luciferase reporter assay
The MYC promoter reporter vector was obtained from Gen-
echem. For luciferase reporter assays, transient infection of reporter
promoters along with a Renilla luciferase control plasmid was fol-
lowed by luciferase activity detection by the Dual-luciferase re-
porter assay instruction (Promega) after 24 h. The experiments
were carried out in duplicates and data were obtained from three
independent experiments.
 X. Gao et al. / Biochemical and Biophysical Research Communications 469 (2016) 679e685
2.9. Chromatin immunoprecipitation (ChIP) assay
The ChIP assay was performed as previously described. The
DNA-protein complexes were washed, and the cross-links were
subsequently reversed at 65
C overnight. RNase 0.2 mg/mL and
proteinase K 0.2 mg/mL were added at 55
C for 2 h to remove RNA
and unbounded proteins. MYC promoter primers for PCR were
purchased from Sangon. The primer sequences for MYC promoter
were: 50 -CCTTTATATTCCGGGGGTCT-30 (forward) and 50 -
CGCTCACTCCCTCTGTCTCT-30 (reverse).
2.10. Tumor xenografts and in vivo treatment
Cells (5  106) were subcutaneously injected into 3e4 weeks old
nude mice (n ¼ 10 per group) in 200 mL PBS per injection in the
right lower extremity. When the tumor sizes were measureable, the
mice were randomly assigned into two groups for drug treatment
trials: (a) JQ1 (50 mg/kg body weight), (b) equal volume of DMSO
(diluent-specific control). Intraperitoneal injections of DMSO or
DMSO-dissolving JQ1were carried out daily. Tumor volume was
determined by the following formula: V ¼ a (length)  b
(width)  c (height)  0.5236. Experiment was terminated after
three weeks and all mice were sacrificed. The tumors were then
excised, weighed, fixed in 4% paraformaldehyde and preserved in
70% ethanol. The maintenance of mice and all experimental pro-
tocols were reviewed and approved by the Animal Care Committee
of Tongji Medical College.
2.11. Statistical analysis
Statistical Package for the Social Sciences (SPSS) software
(version 13.0, SPSS Inc., Chicago, IL, USA) and GraphPad Prism
(Version 5, La Jolla, CA, USA) were applied for data analyses. Paired
t-tests were conducted to determine the difference between paired
tissue specimens by real-time PCR and western blotting analysis.
All tests were two-sided, and statistical significance was set at
P < 0.05.
3. Results
3.1. BRD4 is over-expressed in human DTC tissues and cell lines
To determine BRD4 expression levels in DTC, 45 pairs of human
papillary DTC specimens and surrounding normal thyroid tissues
were obtained. BRD4 mRNA and protein expression levels were up-
regulated in tumor specimens than that in corresponding normal
thyroid tissues (P < 0.05), as demonstrated in Fig. 1 (1A and 1C).
High expression of BRD4 was also detected in papillary DTC K1 and
BCPAP cell lines as compared with Nthy Ori3-1 cell line (Fig. 1B and
D). All these results indicated that BRD4 protein may play roles in
the progression of DTC.
3.2. Inhibition of BRD4 decreased cell viability, induced cell cycle
arrest and enhanced radioiodine uptake in vitro
To investigate whether inhibition of BRD4 could affect the cell
biological activity, we treated K1 and BCPAP cells with different
concentrations of JQ1. After JQ1 treatment, the cell viability was
decreased in both K1 (Fig. 2A) and BCPAP (Fig. 2B) cell lines using
MTT colorimetric assay. Cell cycle assay after JQ1 treatment showed
that cell cycle was arrested at G0/G1 phase compared with the
corresponding negative control (Fig. 2CeE). The cell iodine uptake
ability was subsequently explored. As shown in Fig. 2F and G,
radioiodine uptake was significantly increased after different con-
centrations of JQ1 treatment (P < 0.05) compared with the
681
corresponding control group in both K1 and BCPAP cell lines. After
treatment of 500 nM JQ1, iodine uptake was elevated rapidly in the
first several minutes and then gradually reached the maximal levels
at about 60 min, which was about 2.9 times (Fig. 2H) and 2.6 times
(Fig. 2I) higher than that in control with the DMSO treatment in K1
and BCPAP, respectively (P < 0.05). Iodine uptake after JQ1 treat-
ment along with NaClO4 block was merely reversed compared with
the control group (P > 0.05). Background values obtained from
NaClO4 block group were subtracted. These results demonstrated
that JQ1 influences thyroid cancer cell viability, cell cycle and iodine
uptake in a dose and time dependent manner.
3.3. JQ1 inhibits transcriptional expression of C-MYC and promotes
expression of NIS and P21
Several previous studies have reported that C-MYC could be
repressed by JQ1 in a subset of cancer types, and the expression of
NIS was closely related to iodine uptake. To evaluate whether in-
hibition of BRD4 affects the expression level of C-MYC and NIS in
thyroid cancer, we respectively treated K1 and BCPAP cells with
different concentrations of JQ1. Relative expression levels of BRD4,
C-MYC, NIS and P21 after different JQ1 concentrations and designed
time intervals were examined. As shown in Fig. 3, both mRNA and
protein expression levels of C-MYC but not BRD4 were significantly
down-regulated, while NIS and P21 were up-regulated (Fig. 3AeH),
when compared versus the corresponding control. To further
investigate the relationship between BRD4 and C-MYC, luciferase
assay and ChIP experiments were carried out subsequently.
Decreased promoter activity of C-MYC induced by JQ1 treatment
was illustrated through luciferase reporter system in K1 and BCPAP
cells (Fig. 3I). It was revealed that BRD4 was enriched in the pro-
moter region of C-MYC, and the enrichment of BRD4 in the pro-
moter region of C-MYC could be markedly blocked by JQ1
treatment (Fig. 3J). Thus, BRD4 probably play an important role in
promoting C-MYC expression by binding to its promoter region,
resulting in cell proliferation. Treatment with JQ1 enhances iodine
uptake probably rely on upregulating the expression of NIS.
3.4. BRD4 inhibition impairs tumor growth in vivo
We further evaluated the potential effect of JQ1 on the growth of
DTC xenograft tumors in vivo. As shown in Fig. 4AeC, JQ1 treatment
significantly reduced the tumor weight and tumor size when
compared with the control group (P < 0.05). Meanwhile, the tumor
inhibitory ratio on day 21 was 69.3%. Moreover, compared with the
control group, relative mRNA and protein expression levels of C-
MYC but not BRD4 were significantly down-regulated, whereas NIS
were up-regulated (Fig. 4D and E), which were proved to be
consistent with the results in vitro. These results further demon-
strated that inhibition of BRD4 is an important treatment strategy
to suppress thyroid tumor growth.
4. Discussion
TC is the most common endocrine malignancy, accounting for
almost 1% of all cancers in human populations [16]_ENREF_16.
Nowadays, the profound effects of gene dysregulation on tumor
genesis and cancer progression are becoming evident and are
presently the major subject of research. In thyroid cancer, gene
dysregulation have been reported to contribute to tumor progres-
sion [17]. In addition to the many classical genetic mutations that
have been proved over the last several decades, recent studies
highlight the significance of other epigenetic regulators such as
histone modifiers that are frequently mutated or deregulated in
DTC [18e21] _ENREF_16. As important conserved epigenome
682 X. Gao et al. / Biochemical and Biophysical Research Communications 469 (2016) 679e685

Fig. 1. Over-expression of BRD4 in DTC tissues and cell lines. Relative expression level of BRD4 mRNA from 45 pairs of human DTC specimens (A) and human follicular epithelial cell
line Nthy Ori3-1 and human papillary DTC cell lines K1 and BCPAP (B) were determined by real time PCR, using GAPDH gene as the internal control. Western blot was used to
examine BRD4 protein expression levels in human DTC samples (C) and cell lines of Nthy Ori3-1, K1 and BCPAP (D), using GAPDH as the internal control. *P < 0.05 versus respective
negative control.

Fig. 2. BRD4 inhibition induced cell viability suppression, cell cycle arrest and iodine uptake enhancement. The cell viability of K1 (A) and BCPAP (B) determined by MTT was
performed at designed time intervals (24 h, 48 h and 72 h) after JQ1 (250 nM, 500 nM, 1000 nM) treatment. G0/G1 arrest induced by JQ1 treatment was performed by flow
cytometry (C). Compared with the DMSO treatment, percentages of G0/G1 cell cycle arrest were analyzed for both K1 and BCPAP cells (D, E). Iodine uptake assays were examined
60 min after JQ1 (250 nM, 500 nM, 1000 nM) treatment for both K1 (F) and BCPAP (G) cell lines. Radioiodine uptake tests after treatments of JQ1, NaClO4 block, or JQ1 along with
NaClO4 block were carried out at various time points of radioiodine incubation (H, I). Background values obtained from NaClO4 block group were subtracted. *P < 0.05 versus
respective negative control. Results are the means ± SD in triplicate.
 X. Gao et al. / Biochemical and Biophysical Research Communications 469 (2016) 679e685 683

Fig. 3. Inhibition of BRD4 suppresses C-MYC transcription and enhances NIS expression in vitro. K1 and BCPAP were treated with DMSO or different concentrations of JQ1 (250, 500
or 1000 nM) for 48 h, and the relative mRNA expression levels of BRD4, C-MYC and NIS were analyzed (AeC) by real time PCR. K1 and BCPAP were treated with JQ1 (500 nM) for
indicated time intervals (0, 12, 24 or 48 h) and the mRNA expression levels versus control group were examined (DeF). Western blot was used to monitor relative protein expression
levels of BRD4, C-MYC, NIS and P21 for different concentrations of JQ1 (250, 500 or 1000 nM) or for indicated time intervals (0, 12, 24 or 48 h) (G, H). Luciferase reporter assay
indicated that BRD4 inhibition influenced the C-MYC promoter activity (I). CHIP experiments demonstrated that BRD4 could be recruited to MYC promoter and JQ1 treatment
markly attenuates the recruitment of BRD4 to MYC promoter (J). *P < 0.05 versus respective negative control. Results are the means ± SD in triplicate.

readers, the BET family proteins link to chromatin remodeling, and
generally act as transcription regulators in a series of cells [22,23].
The important BET member BRD4 is aberrantly regulated in several
cancer cells and tissues with unknown mechanism [8,24]. In this
study, we observed that BRD4 was upregulated in DTC cells and
tissues, indicating a pro-oncogenic function of BRD4 in cancer
progression. Moreover, we demonstrated that inhibition of BRD4
using JQ1 influences cell proliferation, cell radioiodine uptake and
tumor growth in K1 and BCPAP cells and mouse models, suggesting
a potential role of BRD4 in regulating DTC biological activity, which
has not been reported.
Traditionally, many patients with well-differentiated subtypes
of TC are cured with the management of removal of the primary
tumor, adjuvant therapies, and TSH suppressive therapy, while
poorly differentiated types usually have unfavorable prognosis [5].
However, clinical trials about molecular therapy exhibited response
rates ranging from 3 to 50%, which was far from satisfactory. Pre-
vious studies have exhibited that pharmacologic inhibition of BET
proteins induces cell growth arrest or cell apoptosis in several
disease models. Evidence of efficacy and safety of the BET inhibitors
in preclinical testing is promoting the rapid development of these
specific compounds for further clinical evaluation. As a first-in-
class BET inhibitor, JQ1 competitively bind to the acetyl-lysine
recognition area of BRD4, interferes with the ability of BRD4 to
identify the acetylated chromatin marks that facilitate gene tran-
scriptional activation [10,25]. The findings that JQ1 exerts anti-
tumor activity in a series of cancer types through suppressing c-
MYC transcription by diminishing the recruitment of BRD4 to the
promoter of c-MYC supply the rationale to evaluate whether
administration of JQ1 influences thyroid cancer growth [10,26,27].
Our results strongly prove that JQ1 enables to inhibit cell prolifer-
ation and tumor growth in thyroid cancer mainly through represses
c-MYC expression relying on BRD4 release from chromosome
without affecting BRD4 expression, which indicates that BET pro-
tein, BRD4, is a novel epigenetic regulator of thyroid cancer and JQ1
may be an innovative therapeutic approach for thyroid cancer
treatment. In addition, it has been reported that C-MYC could
suppress the expression of P21 protein [28,29], the accumulation of
P21 protein induced by JQ1 treatment could at least partly attribute
to the C-MYC inhibition in thyroid cancer cells.
The NIS protein is closely involved in radioiodine uptake, and it
has been widely used for diagnosis and radioiodine treatment of
DTC [30]. Radioiodine can effectively treat DTC foci or metastasis
foci that harboring 131I uptake ability [31]. By contrast, a poor
prognosis is mainly associated with dedifferentiation of DTC
resulted from deregulation of NIS and/or TPO, which are respon-
sible for iodine uptake and the differentiated thyroid phenotype.
Up-regulation of the NIS expression level could effectively enhance
684 X. Gao et al. / Biochemical and Biophysical Research Communications 469 (2016) 679e685
Fig. 4. JQ1 treatment impairs growth of human papillary thyroid xenorafts in vivo. Tumors were weighed after all mice sacrificed (A). Tumor volume curves of JQ1 treatment and
DMSO control (B) were based on the following formula: V ¼ a (length)  b (width)  c (height)  0.5236 and the tumors were photographed (C). The relative mRNA expression
levels of BRD4, C-MYC and NIS of tumor were analyzed by real time PCR (D). Western blot was used to analyze the tumor protein expression levels of BRD4, C-MYC, NIS and P21 (E).
*P < 0.05 versus respective negative control. Results are the means ± SD in triplicate.
the curative effects of radioiodine. In the present study, iodine
uptake and NIS expression in JQ1 treatment group were strikingly
higher than that in control in K1 and BCPAP cell lines. These results
indicated that the enhanced iodine uptake by JQ1 treatment at least
partly attribute to up-regulation of NIS. Our discovery reinforced
the hope of 131I treatment for further clinical application, particu-
larly for patients with 131I refractory thyroid cancers. Future in-
vestigations about the mechanism how JQ1 regulated NIS
expression in DTC will be carried out. It has been reported that a
series of therapeutic strategies influence the treatment effect of
radioiodine therapy in human thyroid cancer cells, whether treat-
ment with JQ1 and 131I simultaneously exert a synergistic effect in
thyroid cancer therapy needs to be verified in the future.
In conclusion, we demonstrate the aberrant expression of BRD4
in thyroid cancers, and provide the possibility of employing BRD4
inhibitors for thyroid cancer treatment. Our study enriches the
knowledge about the therapeutic targets in thyroid cancer pro-
gression and iodine uptake, which is thus likely to enhance the
survival of patients with DTC.
Appendix A. Supplementary data
Supplementary data related to this article can be found at http://
dx.doi.org/10.1016/j.bbrc.2015.12.008.
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