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Tumor Biology |
Laboratoire de Génétique Moléculaire–UPRES JE 2195, Faculté des Sciences Pharmaceutiques et Biologiques de Paris, F-75006 Paris [I. B., B. P., M. O., M. V.]; Laboratoire d’Oncogénétique-Institut National de la Santé et de la Recherche Médicale (INSERM) E0017 [I. B., R. L.] and Laboratoire d’Anatomo-Cytopathologie [V. L. D.], Centre René Huguenin, F-92211 St-Cloud; and Institut de Génétique et de Biologie Moléculaire et Cellulaire-INSERM U184, Université Louis Pasteur, F-67404 Illkirch Cedex [M-C. R.], France
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ABSTRACT |
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 Top ABSTRACT INTRODUCTIONMATERIALS AND METHODSImmunohistochemical StudiesStatistical AnalysisRESULTSDISCUSSIONREFERENCES |
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Here, we identified the well-known CGA gene (coding for
the 
subunit of glycoprotein hormones) as a new ER-responsive
gene in human breast cancer cells. We used a real-time quantitative
reverse transcription-PCR assay to quantify CGA mRNA
copy numbers in a large series of breast tumors. CGA
overexpression (>10 SD above the mean for normal breast
tissues) was observed in 44 of 131 (33.6%) breast tumor RNAs, ranging
from 20 to 16,500 times the level in normal breast tissues; the highest
levels of CGA gene expression were close to those
observed in placenta.
Significant links were observed between CGA gene
overexpression and Scarff-Bloom-Richardson histopathological grade I+II
(P = 0.015), and progesterone
(P = 0.0009) and estrogen
(P < 10-7) receptor
positivity, which suggested that CGA is a marker of low
tumor aggressiveness. We observed CGA mRNA
overexpression in 44 of 90 (48.9%) ER-positive tumors and in none
of the 41 ER-negative tumors. Immunohistochemical studies
demonstrated that human chorionic gonadotropin protein was strictly
limited to ER-positive tumor cells. Overexpression of the
CGA gene was not accompanied by overexpression of the
CGB gene.
Our results also suggest that CGA could be a more
reliable marker than PS2 and PR for ER
functionality and, thus, for endocrine responsiveness. Moreover, the
CGA marker has the added value of dichotomizing
ER-positive patients into two subgroups of similar size. Specific
antibodies directed to secreted human chorionic gonadotropin
protein are commercially available, thus facilitating the future
application of this marker to the clinical management of breast cancer.
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INTRODUCTION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSImmunohistochemical StudiesStatistical AnalysisRESULTSDISCUSSIONREFERENCES |
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We fortuitously identified the well-known CGA gene (coding
for the subunit of glycoprotein hormones) as a new ER-responsive
gene in human breast cancer cells. Indeed, the initial purpose of this
study was to determine whether the emergence of "trophoblastic"
CGB genes, which we have previously shown to be associated
with malignant breast transformation (4)
, was accompanied
by cooverexpression of the CGA gene and the production of
ectopic hCG heterodimeric hormone in breast tumor cells. We thus
quantified CGA and CGB gene expression in the
large series of unilateral invasive primary breast tumor RNAs by means
of real-time quantitative RT-PCR assays based on TaqMan methodology. We
unexpectedly found marked differences in the total amount of
CGA mRNA molecules that were clearly independent of
CGB expression status but fully associated with the
ER-positive phenotype in our breast tumor series. Indeed,
CGA gene overexpression was observed in one-half of the
ER-positive tumors and in none of the ER-negative tumors.
Immunohistochemical studies confirmed the hCG protein overexpression
and showed that it was strictly limited to ER-positive tumor cells.
To further examine the unexpected link between CGA gene
overexpression and ER positivity, we analyzed the expression of the
estrogen-responsive genes PR and PS2. Our results
suggest that CGA could be a more reliable marker than
PR and PS2 for ER functionality and, thus, for
endocrine responsiveness.
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MATERIALS AND METHODS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSImmunohistochemical StudiesStatistical AnalysisRESULTSDISCUSSIONREFERENCES |
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Immunohistochemical study was performed on fixed, paraffin-embedded tissue sections from 20 of the 131 breast tumor specimen.
The patients (mean age, 58.2 years; range, 34–91) met the following
criteria: primary unilateral nonmetastatic breast carcinoma on which
complete clinical, histological, and biological data were available;
and no radiotherapy or chemotherapy before surgery. The main prognostic
factors are presented in Table 1
. The median follow-up was 8.1 years (range, 1.0–15.9). Forty-seven
patients relapsed (the distribution of first relapse events was as
follows: 13 local and/or regional recurrences, 30 metastases, and 4
both).
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Evaluation of "Classical" Prognostic Factors
The histological type and steroid-hormone receptor status of
each tumor, and the number of positive axillary nodes, were established
at the time of surgery. The malignancy of infiltrating carcinomas was
scored according to Bloom and Richardson’s histoprognostic system
(5)
. ER and PR status was assayed by biochemical method
(EIA) as described by the European Organization for Research and
Treatment of Cancer (6)
, with a detection limit of 10
fmol/mg cytosolic protein.
Real-Time RT-PCR
Theoretical Basis.
Quantitative values are obtained from the threshold cycle number at
which the increase in the signal associated with exponential growth of
PCR products begins to be detected using PE Biosystems analysis
software, according to the manufacturer’s manuals.
The precise amount of total RNA added to each reaction mix (based on absorbance) and its quality (i.e., lack of extensive degradation) are both difficult to assess. We, therefore, also quantified transcripts of the RPLP0 gene (also known as 36B4) encoding human acidic ribosomal phosphoprotein P0 as the endogenous RNA control, and each sample was normalized on the basis of its RPLP0 content.
The relative target gene expression level was also normalized to a calibrator, or 1x sample, consisting of a normal breast tissue sample (quantitation of CGA, CGB, and RPLP0 genes) or of the breast tumor tissue sample that contained the smallest amount of target gene mRNA (quantitation of other target genes).
Final results, expressed as N-fold differences in target
gene expression relative to the RPLP0 gene and the
calibrator, termed Ntarget, were determined as
follows:
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Ct values of the sample and
calibrator are determined by subtracting the average Ct
value of the target gene from the average Ct value of the
RPLP0 gene.
Primers, Probes, and PCR Consumables.
Primers and probes for the RPLP0 and target genes were
chosen with the assistance of the computer programs Oligo 4.0 (National
Biosciences, Plymouth, MN) and Primer Express (Perkin-Elmer Applied
Biosystems, Foster City, CA). We conducted BLASTN searches against
dbEST, htgs, and nr (the nonredundant set of the
GenBank, European Molecular Biology Laboratory, and DNA
Data Bank of Japan database sequences) to confirm the total gene
specificity of the nucleotide sequences chosen for the primers and
probes, and the absence of DNA polymorphisms. The nucleotide sequences
of the oligonucleotide hybridization probes and primers are shown in
Table 2
. To avoid amplification of contaminating genomic DNA, one of the two
primers was placed in a different exon. For example, the upper primer
and the probe of CGA were placed in exon 3, and the lower
primer was placed at the junction between exons 3 and 4.
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cDNA Synthesis.
RNA was reverse transcribed in a final volume of 20 µl containing 1x
RT buffer [500 mM each dNTP, 3 mM MgCl2, 75
mM KCl, and 50 mM Tris-HCl (pH 8.3)], 10 units
of RNasinTM RNase inhibitor (Promega, Madison, WI), 10 mM
DTT, 50 units of Superscript II RNase H-reverse transcriptase (Life
Technologies, Inc., Gaithersburg, MD), 1.5 mM random
hexamers (Pharmacia, Uppsala, Sweden), and 1 µg of total RNA. The
samples were incubated at 20°C for 10 min and 42°C for 30 min, and
reverse transcriptase was inactivated by heating at 99°C for 5 min
and cooling at 5°C for 5 min.
PCR Amplification.
All of the PCR reactions were performed using an ABI Prism 7700
Sequence Detection system (Perkin-Elmer Applied Biosystems). PCR was
performed using either the TaqMan PCR Core Reagents kit or the SYBR
Green PCR Core Reagents kit (Perkin-Elmer Applied Biosystems). The
thermal cycling conditions comprised an initial denaturation step at
95°C for 10 min and 50 cycles at 95°C for 15 s and 65°C for
1 min. Experiments were performed with duplicates for each data point.
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Immunohistochemical Studies |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSImmunohistochemical StudiesStatistical AnalysisRESULTSDISCUSSIONREFERENCES |
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subunit of glycoprotein hormones [a kind gift from Dr Jean-Michel
Bidart (Service de Biologie Oncologique, Institut Gustave Roussy, 94805
Villejuif Cedex, France); Ref. 7
] and monoclonal antibody
ID5 raised against human ER (Dako SA, Trappes, France). The immunohistochemical procedure was performed on paraffin-embedded tissue sections. A microwave antigen-retrieval technique was used in all of the cases. Sections were mounted on precoated slides (triethylenethiophosphoramide-coated slides) and allowed to dry at 50°C overnight. The sections were then dewaxed in xylene and hydrated through graded concentrations of alcohol. Endogenous activity was blocked with 1% hydrogen peroxide for 15 min. Sections were then immersed in a thermoresistant plastic box containing 10 ml of citrate buffer (pH 6.0) and processed in the microwave oven four times for 5 min each at 750 W. Sections were then allowed to cool at room temperature for 30 min before rinsing in TRIS-buffered saline. The blocking reagent was topped off, and the primary antibodies were left for 1 h. A standard avidin-biotin-peroxidase complex was used to reveal the antibody-antigen reaction.
Localization and intensity of staining were assessed by two pathologists, blinded to the real-time RT-PCR results.
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Statistical Analysis |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSImmunohistochemical StudiesStatistical AnalysisRESULTSDISCUSSIONREFERENCES |
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Clinical, histological, and biological parameters were compared using
the 
2 test. Differences between the two
populations were judged significant at confidence levels greater than
95% (P < 0.05). Survival distributions were
estimated by the Kaplan-Meier method (8)
, and the
significance of differences between survival rates was determined using
the log-rank test.
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RESULTS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSImmunohistochemical StudiesStatistical AnalysisRESULTSDISCUSSIONREFERENCES |
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CGA mRNA Level in Tumor Breast Tissues.
Among the 131 breast tumor RNA samples tested, 44 (33.6%) showed
CGA gene overexpression. Very large differences in the
amount of CGA mRNA were observed (NCGA, from 20.6
to 16500): 10 tumors had an expression level of 20–40 times; 11 tumors
41–100 times; 13 tumors 101–1000 times; and 10 tumors >1000 times
that of normal breast tissue RNA. The highest levels of CGA
gene expression in breast tumors were close to those observed in
placenta (data not shown). CGA gene expression was also
investigated in nine patients from whom both primary breast tumors and
matched normal breast tissue were available: four tumors showed clearly
higher CGA expression in the tumor (NCGA: 2260.1,
70.4, 66.9, and 36.9, respectively) than in the normal tissue
(NCGA: 0.9, 3.4, 2.2, and 1.3, respectively).
Correlation between CGA mRNA Levels and Clinical and
Pathological Parameters.
We then sought links between quantitative CGA mRNA status
and standard clinical, pathological, and biological factors in breast
cancer (Table 3)
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Patients with tumors overexpressing CGA did not relapse more
(or less) frequently (Table 3)
and did not have significantly shorter
(or longer) disease-free survival after surgery compared with patients
whose tumors normally expressed CGA (log-rank test, not
shown).
Localization of hCG Subunit in Tumor Cell Cytoplasm.
Of the 20 tumors studied by immunohistochemistry, we detected
specific immunoreactivity in the 10 tumors that overexpressed
CGA mRNA and in none of the tumors that showed no
CGA overexpression. We thus observed a perfect match between
CGA mRNA overexpression and immunohistochemical
positivity (Fig. 1)
. hCG immunoreactivity was exclusively found inside ER-positive
tumor cells; infiltrating lymphocytes and normal glandular cells in the
tumor were negative. Tumor cell staining was found exclusively in the
cytoplasm, and the staining pattern consisted mainly of isolated
positive cells (Fig. 1)
. The positive cells were chromogranin
A-negative (data not shown).
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. Neither were links observed between CGB gene
overexpression and PR and ER status (data not shown).
Relationship between CGA mRNA Levels and
CCND1, MYC, and ERBB2
Expression Status.
The 131 tumors studied for CGA expression had previously
been tested for mRNA expression of the CCND1,
MYC, and ERBB2 genes (9
, 10)
.4
We found a significant link between CGA overexpression and
CCND1 overexpression (P = 0.00003)
but no link between CGA overexpression and altered mRNA
expression of the other two genes (Table 3)
.
Relationship between CGA mRNA Levels and
PS2 and ERß Expression Status.
To study the link between CGA gene overexpression and ER
positivity, we also quantified, by means of a real-time quantitative
RT-PCR assay, the expression of the ER-responsive gene
PS2, as well as the ER ß (ERß) gene. We also
analyzed mRNA levels of the PR and ER genes to
confirm PR and ER protein status determined by a biochemical (EIA)
method, to obtain quantitative PR and ER
values and to avoid a possible discrepancy due to tumor heterogeneity.
As regards the ER, PS2, ERß, and
PR markers, patients were subdivided into three equal groups
of tumors with low (n = 43), intermediate
(n = 44), and high (n = 44) mRNA levels. A total correlation was observed between
ER gene status determined with biochemical and real-time
RT-PCR. Indeed, all of the 41 ER-negative tumors in the biochemical
(EIA) method had low ER mRNA levels. Then, breast tumors
were subdivided into ER-negative tumors (low
ER mRNA; n = 43) and
ER-positive tumors (intermediate/high ER
mRNA; n = 88).
The results (summarized in Tables 4
and 5
) showed a stronger association between ER gene status and
CGA gene status (total link) than between ER
gene status and PS2, PR or ERß gene
status (subtotal links). Indeed, several ER-negative (low
ER mRNA expressed) tumors overexpressed PS2
and/or PR, whereas none of the
"ER-negative" tumors overexpressed the CGA
gene. Interestingly, we observed a negative correlation between
ER and ERß mRNA expression levels
(P = 0.0013).
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-positive subgroup
(n = 88), we found a significant link between
CGA overexpression and CCND1 overexpression
(P = 0.0024), but no link between
CGA mRNA expression status and PS2,
PR, or ERß mRNA expression levels (Table 6)
.
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DISCUSSION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSImmunohistochemical StudiesStatistical AnalysisRESULTSDISCUSSIONREFERENCES |
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protein overexpression and showed that it was
strictly limited to breast tumor cells. Our results are in keeping with
a report of hCG protein production by breast tumor cells, as shown
by immunohistochemistry analysis (12)
.
We sought to determine whether the observed overexpression of hCG
protein in breast cancer has to aim to associate with the hCGß
subunit to produce the heterodimeric hormone hCG, which could mediate
biological activities through the luteinizing hormone/hCG
receptor, which was recently shown to be expressed in breast cancer
(13
, 14)
. Although we observed large differences in the
amount of CGB mRNA molecules (as high as 192 times the level
in normal breast tissues), we found no link between CGA mRNA
overexpression and CGB mRNA overexpression in our series of
breast tumors (Table 3)
. We also observed no link between
CGA overexpression and mRNA expression of the
LHB, FSHB, and TSHB genes, which code
for the other three ß subunits capable of associating with the hCG
subunit to produce glycoprotein hormones (data not shown). The highest
levels of CGA gene expression in our breast tumor series,
which were close to those observed in placenta and were much higher
than the amount of CGB mRNA in breast tumors
(>103
-fold lower that placental levels) suggest
that different genetic (or epigenetic) mechanisms are responsible for
CGA and CGB gene overexpression in breast tumors.
Taken together, these results suggest that if the free hCG subunit
(and not the heterodimeric hormone hCG) is involved in breast
tumorigenesis, it would act via a novel pathway not involving the
luteinizing hormone/hCG receptor.
Little is known of the involvement of the hCG subunit itself in
carcinogenic processes. Rivera et al. (15)
showed that an antisense sequence to the CGA gene inhibited
the growth and reversed the transformed phenotype of human lung tumor
cells. One possibility, in breast tissue, is that the hCG subunit
controls PRL (prolactin) gene expression, as in myometrial,
decidual, and pituitary gland cells (16, 17, 18)
. Because
PRL (prolactin) gene expression is altered in breast cancer
(19)
, we tested this hypothesis. We found no link between
CGA mRNA overexpression and PRL mRNA
overexpression, ruling out a role of the CGA gene in
PRL pathway dysregulation in breast tumors (data not shown).
The most important result of our study is that CGA gene
overexpression is perfectly linked to ER positivity in breast tumors.
Indeed, CGA gene overexpression was observed in one-half of
the ER-positive tumors and in none of the ER-negative tumors. We
also found a weaker association between CGA gene
overexpression and PR positivity and SBR histopathological grade I+II,
which suggests that CGA is a marker of tumors with low
aggressiveness. No relationship between ER positivity and
CGB overexpression was observed in the same series. Our
immunohistochemical data confirm that hCG protein overexpression is
strictly limited to ER-positive tumor cells. This makes CGA
status an attractive candidate as a molecular marker, and one that may
prove far more reliable than ER status alone, to predict endocrine
responsiveness. Indeed, only one-half of all of the ER-positive
patients respond favorably to antiestrogen treatment (1)
,
and we observed CGA gene overexpression in one-half of the
ER-positive tumors in our breast tumor series. Moreover,
we observed no link between CGA mRNA expression status and
PS2 or PR mRNA expression levels in the
ER-positive subgroup, which suggests that CGA activation
requires ER but involves a mechanism different from that of
PS2 and PR. Moreover, our results (summarized in
Tables 4
and 5
) suggest that the CGA gene may be a more
reliable marker than PS2 and PR for ER
functionality and, thus, for endocrine responsiveness. Indeed, several
"ER-negative" (low ER mRNA expressed)
tumors overexpressed PS2 and/or PR, whereas none
of the ER-negative tumors overexpressed the
CGA gene. Taken together, these findings suggest that the
CGA marker provides new, independent information that, in
conjunction with ER status, may help to determine the
likelihood that a given tumor will respond to endocrine therapy. We
infer that the subpopulation of CGA-positive,
ER-positive tumors would correspond to those responsive
to hormone therapy. Because endocrine therapy was rarely used 20 years
ago, which is when our samples were obtained, we were unable to study
the relationship between CGA gene overexpression and the
response to endocrine therapy. However, among the 44 ER-positive
patients who received postoperative hormone therapy, 71.4% (15 of 21)
of those who had CGA overexpression are relapse-free,
compared with 43.5% (10 of 23) of those who had normal CGA
expression (P = 0.06).
In addition to their clinical application, our results raise an
important question concerning the mechanisms that lead to
CGA overexpression in ER-positive breast tumors. The
structure of the CGA promoter has been extensively studied
(reviewed in Ref. 20
). Most studies have focused on
CGA gene expression in placenta and pituitary tissues and
suggest that the tissue-specific expression of the CGA gene
is determined by combinations of several elements, some of which are
common to all of the tissues, whereas others are specific to a given
tissue. These studies identified a remarkable series of composite
regulatory elements that interact with families of transcription
factors that are still being characterized. These elements include a
trophoblast-specific element (TSE) that binds TSEB and two CREs that
bind CREBs. However, no consensus ERE was found in the 1638 bp
of the cloned promoter region of the human CGA gene (GenBank
accession no. AF109152). It is noteworthy that we observed no link
between overexpression of the CGA gene and the
PS2 or PR genes, two ER-inducible genes with
ERE elements, which suggests that ER activates CGA gene
expression in breast tumors via a mechanism that does not involve an
ERE element.
The most plausible hypothesis is that CGA gene activation by
ER protein occurs via the CRE element. Indeed, estrogen activates
its receptor by inducing a conformational change in the hormone-binding
domain (21)
, thereby creating a structural motif that
binds to a number of transcriptional coactivators such as SRC-1
(steroid receptor coactivator), p300/CBP (CREB-binding protein), and
P/CAF (p300/CBP-associated factor). Interestingly, p300/CBP can bind to
CREB, which is known to bind to the CRE element, whereas P/CAF is an
enzyme known to promote transcription by acetylating histones, thereby
modifying the chromatin structure and facilitating gene expression.
In this respect it is noteworthy that the CCND1 gene, the
overexpression of which is also strongly linked to ER positivity in
breast tumors, contains a CRE element but not an ERE element in its
promoter. Sabbah et al. (22)
recently showed
that it is this CRE element that is involved in the activation of the
CCND1 gene by ER. Interestingly, we observed a strong
link between CGA mRNA overexpression and CCND1
mRNA overexpression in our breast tumor series (Table 3)
and especially
in the ER-positive subgroup (Table 6)
.
Keri et al. (23)
, using 1500 bp of the proximal
5' flanking sequence of the human CGA gene in in vitro
(cultured cells) or in vivo (transgenic mice) models, did
not observe up-regulation by estrogen (or ER) of reporter gene
expression. We observed no CGA overexpression in the ER-positive
breast cell lines tested (MCF7, T-47D, ZR-75–1) either before or after
estradiol treatment. Taken together, these results suggest that the
mechanism that leads in vivo to CGA overexpression in human
ER-positive breast tumors involves several factors, including ER
and several uncharacterized transcriptional coactivators that are not
all present in the classical in vitro and in vivo
models used to date.
In conclusion, we have identified CGA as a major new
ER-responsive gene that should greatly contribute to our
understanding of the role of the ER in breast cancer. Our results also
make the CGA gene an attractive alternative molecular marker
to PR and PS2 as a refined predictor of endocrine
responsiveness in breast cancer patients. We are currently conducting a
large study of specific breast cancer patient phenotypes to test the
practical value of CGA as a predictor of the response to
hormone therapy. The CGA gene encodes a well-characterized
secreted protein for which specific antibodies are already commercially
available, thus facilitating the future application of this marker to
the clinical management of breast cancer.
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ACKNOWLEDGMENTS |
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e Huguenin, 92211 St-Cloud, France) for helpful contributions.
We are also indebted to Drs. Marie-France Pichon (Service de
Médecine Nucléaire, Centre Ree Huguenin, 92211
St-Cloud, France) and Jean-Michel Bidart (Service de Biologie
Oncologique, Institut Gustave Roussy, 94805 Villejuif Cedex, France)
for kindly providing a number of tissue specimens and the monoclonal
antibody HT13, respectively. We thank Odile Champy for excellent
technical assistance. |
FOOTNOTES |
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1 This work was supported by the Association pour
la Recherche sur le Cancer and the Ministère de l’Enseignement
Supérieur et de la Recherche. 
2 To whom requests for reprints should be
addressed, at Laboratoire de Génétique
Moléculáire-VPRES JE 2195, Faculté des Sciences
Pharmaceutiques et Biologiques de Paris, 4, avenue de
l’Observatoire, 75006 Paris, France. Phone: 33-1-53-73-97-25; Fax:
33-1-44-07-17-54; E-mail: mvidaud{at}teaser.fr
3 The abbreviations used are: ER, estrogen
receptor; PR, progesterone receptor; hCG, human chorionic gonadotropin;
RT-PCR, reverse transcription-PCR; EIA, enzyme immunoassay;
SBR, Scarff-Bloom-Richardson; ERE, estrogen-response element; CRE, cAMP
response element; CREB, CRE binding protein; CBP, CREB binding
protein.
4 I. Bièche, M. Olivi, C. Noguès, M.
Vidaud, and R. Lidereau. Prognostic value of CCND1 gene
status in sporadic breast tumors, as determined by real-time
quantitative PCR assays, manuscript in preparation.
Received 6/ 6/00. Accepted 12/14/00.
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REFERENCES |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSImmunohistochemical StudiesStatistical AnalysisRESULTSDISCUSSIONREFERENCES |
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-subunit. I. Characterization of topographic sites recognized by monoclonal antibodies. J. Biol. Chem., 263: 10364-10369, 1988.-subunit HCG demonstrated in breast carcinomas by the immunoperoxidase technique.. J. Clin. Pathol., 31: 245-249, 1978. subunit of human chorionic gonadotropin.. J. Cell Biol., 108: 2423-2434, 1989. subunit.. Science (Washington DC), 226: 566-568, 1984.-subunit in vitro.. Fertil. Steril., 64: 972-976, 1995.[Medline]
molecules from pregnancy stimulate secretion of prolactin from human decidual cells: a novel function for free in pregnancy.. Endocrinology, 129: 2257-2259, 1991.-subunit gene despite the absence of a high affinity binding site for estrogen receptor.. Mol. Endocrinol., 5: 725-733, 1991.This article has been cited by other articles:
|
|
 |
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  A. Duces, R. Karaky, D. Martel-Renoir, L. Mir, Y. Hamma-Kourbali, I. Bieche, P. Opolon, J. Delbe, J. Courty, M. Perricaudet, et al. 16-kDa fragment of pleiotrophin acts on endothelial and breast tumor cells and inhibits tumor development Mol. Cancer Ther., September 1, 2008; 7(9): 2817 - 2827. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
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|
|
|
|
 |
|
 F. Zemani, J.-S. Silvestre, F. Fauvel-Lafeve, A. Bruel, J. Vilar, I. Bieche, I. Laurendeau, I. Galy-Fauroux, A. M. Fischer, and C. Boisson-Vidal Ex Vivo Priming of Endothelial Progenitor Cells With SDF-1 Before Transplantation Could Increase Their Proangiogenic Potential Arterioscler Thromb Vasc Biol, April 1, 2008; 28(4): 644 - 650. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Fritah, C. Saucier, O. De Wever, M. Bracke, I. Bieche, R. Lidereau, C. Gespach, S. Drouot, G. Redeuilh, and M. Sabbah Role of WISP-2/CCN5 in the Maintenance of a Differentiated and Noninvasive Phenotype in Human Breast Cancer Cells Mol. Cell. Biol., February 1, 2008; 28(3): 1114 - 1123. [Abstract] [Full Text] [PDF]
|
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|
 |
|
 C. Le Stunff, A. Dechartres, V. Mariot, C. Lotton, C. Trainor, E. Miraglia Del Giudice, D. Meyre, I. Bieche, I. Laurendeau, P. Froguel, et al. Association Analysis Indicates That a Variant GATA-Binding Site in the PIK3CB Promoter Is a Cis-Acting Expression Quantitative Trait Locus for This Gene and Attenuates Insulin Resistance in Obese Children Diabetes, February 1, 2008; 57(2): 494 - 502. [Abstract] [Full Text] [PDF]
|
|
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|
 |
|
 I. Bieche, C. Chavey, C. Andrieu, M. Busson, S. Vacher, L. Le Corre, J.-M. Guinebretiere, S. Burlinchon, R. Lidereau, and G. Lazennec CXC chemokines located in the 4q21 region are up-regulated in breast cancer Endocr. Relat. Cancer, December 1, 2007; 14(4): 1039 - 1052. [Abstract] [Full Text] [PDF]
|
|
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|
 |
|
 D. C. Lelong, I. Bieche, E. Perez, K. Bigot, J. Leemput, I. Laurendeau, M. Vidaud, J.-P. Jais, M. Menasche, and M. Abitbol Novel Mouse Model of Monocular Amaurosis Fugax Stroke, December 1, 2007; 38(12): 3237 - 3244. [Abstract] [Full Text] [PDF]
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|
 S Tozlu-Kara, V Roux, C Andrieu, J Vendrell, S Vacher, V Lazar, F Spyratos, M Tubiana-Hulin, P Cohen, P Dessen, et al. Oligonucleotide microarray analysis of estrogen receptor {alpha}-positive postmenopausal breast carcinomas: identification of HRPAP20 and TIMELESS as outstanding candidate markers to predict the response to tamoxifen J. Mol. Endocrinol., October 1, 2007; 39(4): 305 - 318. [Abstract] [Full Text] [PDF]
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J. T. Buijs, N. V. Henriquez, P. G.M. van Overveld, G. van der Horst, I. Que, R. Schwaninger, C. Rentsch, P. ten Dijke, A.-M. Cleton-Jansen, K. Driouch, et al. Bone Morphogenetic Protein 7 in the Development and Treatment of Bone Metastases from Breast Cancer Cancer Res., September 15, 2007; 67(18): 8742 - 8751. [Abstract] [Full Text] [PDF]
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D. Marot, I. Bieche, C. Aumas, S. Esselin, C. Bouquet, S. Vacher, G. Lazennec, M. Perricaudet, F. Kuttenn, R. Lidereau, et al. High tumoral levels of Kiss1 and G-protein-coupled receptor 54 expression are correlated with poor prognosis of estrogen receptor-positive breast tumors Endocr. Relat. Cancer, September 1, 2007; 14(3): 691 - 702. [Abstract] [Full Text] [PDF]
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E. Pasmant, I. Laurendeau, D. Heron, M. Vidaud, D. Vidaud, and I. Bieche Characterization of a Germ-Line Deletion, Including the Entire INK4/ARF Locus, in a Melanoma-Neural System Tumor Family: Identification of ANRIL, an Antisense Noncoding RNA Whose Expression Coclusters with ARF Cancer Res., April 15, 2007; 67(8): 3963 - 3969. [Abstract] [Full Text] [PDF]
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 P. Levy, H. Ripoche, I. Laurendeau, V. Lazar, N. Ortonne, B. Parfait, K. Leroy, J. Wechsler, I. Salmon, P. Wolkenstein, et al. Microarray-Based Identification of Tenascin C and Tenascin XB, Genes Possibly Involved in Tumorigenesis Associated with Neurofibromatosis Type 1 Clin. Cancer Res., January 15, 2007; 13(2): 398 - 407. [Abstract] [Full Text] [PDF]
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S Tozlu, I Girault, S Vacher, J Vendrell, C Andrieu, F Spyratos, P Cohen, R Lidereau, and I Bieche Identification of novel genes that co-cluster with estrogen receptor alpha in breast tumor biopsy specimens, using a large-scale real-time reverse transcription-PCR approach Endocr. Relat. Cancer, December 1, 2006; 13(4): 1109 - 1120. [Abstract] [Full Text] [PDF]
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 C. Sevin, A. Benraiss, D. Van Dam, D. Bonnin, G. Nagels, L. Verot, I. Laurendeau, M. Vidaud, V. Gieselmann, M. Vanier, et al. Intracerebral adeno-associated virus-mediated gene transfer in rapidly progressive forms of metachromatic leukodystrophy Hum. Mol. Genet., January 1, 2006; 15(1): 53 - 64. [Abstract] [Full Text] [PDF]
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 I. Girault, N. Rougier, C. Chesne, R. Lidereau, P. Beaune, I. Bieche, and I. de Waziers SIMULTANEOUS MEASUREMENT OF 23 ISOFORMS FROM THE HUMAN CYTOCHROME P450 FAMILIES 1 TO 3 BY QUANTITATIVE REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION Drug Metab. Dispos., December 1, 2005; 33(12): 1803 - 1810. [Abstract] [Full Text] [PDF]
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 J. Arredondo, A. I. Chernyavsky, A. Karaouni, and S. A. Grando Novel Mechanisms of Target Cell Death and Survival and of Therapeutic Action of IVIg in Pemphigus Am. J. Pathol., December 1, 2005; 167(6): 1531 - 1544. [Abstract] [Full Text] [PDF]
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D. M. Smadja, I. Bieche, G. Uzan, H. Bompais, L. Muller, C. Boisson-Vidal, M. Vidaud, M. Aiach, and P. Gaussem PAR-1 Activation on Human Late Endothelial Progenitor Cells Enhances Angiogenesis In Vitro With Upregulation of the SDF-1/CXCR4 System Arterioscler Thromb Vasc Biol, November 1, 2005; 25(11): 2321 - 2327. [Abstract] [Full Text] [PDF]
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M. Asheuer, I. Bieche, I. Laurendeau, A. Moser, B. Hainque, M. Vidaud, and P. Aubourg Decreased expression of ABCD4 and BG1 genes early in the pathogenesis of X-linked adrenoleukodystrophy Hum. Mol. Genet., May 15, 2005; 14(10): 1293 - 1303. [Abstract] [Full Text] [PDF]
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 M.-C. Lacroix, J. Guibourdenche, T. Fournier, I. Laurendeau, A. Igout, V. Goffin, J. Pantel, V. Tsatsaris, and D. Evain-Brion Stimulation of Human Trophoblast Invasion by Placental Growth Hormone Endocrinology, May 1, 2005; 146(5): 2434 - 2444. [Abstract] [Full Text] [PDF]
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J. Arredondo, A. I. Chernyavsky, L. M. Marubio, A. L. Beaudet, D. L. Jolkovsky, K. E. Pinkerton, and S. A. Grando Receptor-Mediated Tobacco Toxicity: Regulation of Gene Expression through {alpha}3{beta}2 Nicotinic Receptor in Oral Epithelial Cells Am. J. Pathol., February 1, 2005; 166(2): 597 - 613. [Abstract] [Full Text] [PDF]
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 S. F. Fernandez, M.-H. Huang, B. A. Davidson, P. R Knight III, and J. L. Izzo Jr Mechanisms of Angiotensin II-Mediated Decreases in Intraneuronal Ca2+ in Calcium-Loaded Stellate Ganglion Neurons Hypertension, February 1, 2005; 45(2): 276 - 282. [Abstract] [Full Text] [PDF]
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I. Bieche, F. Lerebours, S. Tozlu, M. Espie, M. Marty, and R. Lidereau Molecular Profiling of Inflammatory Breast Cancer: Identification of a Poor-Prognosis Gene Expression Signature Clin. Cancer Res., October 15, 2004; 10(20): 6789 - 6795. [Abstract] [Full Text] [PDF]
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P. Levy, I. Bieche, K. Leroy, B. Parfait, J. Wechsler, I. Laurendeau, P. Wolkenstein, M. Vidaud, and D. Vidaud Molecular Profiles of Neurofibromatosis Type 1-Associated Plexiform Neurofibromas: Identification of a Gene Expression Signature of Poor Prognosis Clin. Cancer Res., June 1, 2004; 10(11): 3763 - 3771. [Abstract] [Full Text] [PDF]
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 J. Alberola, A. Rodriguez, O. Francino, X. Roura, L. Rivas, and D. Andreu Safety and Efficacy of Antimicrobial Peptides against Naturally Acquired Leishmaniasis Antimicrob. Agents Chemother., February 1, 2004; 48(2): 641 - 643. [Abstract] [Full Text] [PDF]
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 I. Bieche, A. Laurent, I. Laurendeau, L. Duret, Y. Giovangrandi, J.-L. Frendo, M. Olivi, J.-L. Fausser, D. Evain-Brion, and M. Vidaud Placenta-Specific INSL4 Expression Is Mediated by a Human Endogenous Retrovirus Element Biol Reprod, April 1, 2003; 68(4): 1422 - 1429. [Abstract] [Full Text] [PDF]
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 A. Yoshida, N. Suzuki, Y. Nakano, T. Oho, M. Kawada, and T. Koga Development of a 5' Fluorogenic Nuclease-Based Real-Time PCR Assay for Quantitative Detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis J. Clin. Microbiol., February 1, 2003; 41(2): 863 - 866. [Abstract] [Full Text] [PDF]
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A. Dallol, N. F. Da Silva, P. Viacava, J. D. Minna, I. Bieche, E. R. Maher, and F. Latif SLIT2, a Human Homologue of the Drosophila Slit2 Gene, Has Tumor Suppressor Activity and Is Frequently Inactivated in Lung and Breast Cancers Cancer Res., October 15, 2002; 62(20): 5874 - 5880. [Abstract] [Full Text] [PDF]
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 S. Bretagne, R. Durand, M. Olivi, J.-F. Garin, A. Sulahian, D. Rivollet, M. Vidaud, and M. Deniau Real-Time PCR as a New Tool for Quantifying Leishmania infantum in Liver in Infected Mice Clin. Vaccine Immunol., July 1, 2001; 8(4): 828 - 831. [Abstract] [Full Text] [PDF]
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