Download Motility-related Protein-1 (MRP-1/CD9) Reduction as a Factor of

ICANCER RESEARCH 56. 1244-1249,
March 15. 1996
Advances in Brief
Motility-related Protein-1 (MRP-1/CD9) Reduction as a Factor of Poor Prognosis in
Breast Cancer1
Masayuki Miyake,2 Keiji Nakano, Shin-ichi Itoi, Takashi Koh, and Toshihiko Taki
Department of Thoracic Surgery and Department V of Oncology. Kitano Hospital, Tazitke Koftikai Medical Research Insiliate. 13-3. Katni\tiittacho.
IM. M.. K. N.. S-i. !.. T. T.j. and Department of Pathology and Tumor Biology. Faculty of Medicine. Kyoto University, Kyoto 606 ¡T.KJ, Japan
Abstract
The application of reliable markers is of major importance for predict
ing the prognosis of and instituting the appropriate postsurgical treatment
of patients with breast cancer. Previously we showed that motility-related
[unfein-1 iMKI'-l i. which is identical to CD9, regulates cell motility, and
that cultured tumor cells transfected with MRP-1/CD9 cDNA have low
motility and low mctastatic potential. In addition, MKP-1/CD9 immunoblotting and immunohistochemical study with breast cancer revealed that
MRP-1/CD9 expression diminished as the clinical stage of a given breast
cancer advanced and that the MRP-1/CD9 gene and protein expression in
the metastatic lymph nodes was strikingly lower than in the primary
breast cancers. In this study, we also investigated the expression of MRP1/CD9 by immunoblotting and immunohistochemical
analysis in 143
freshly resected invasive ductal carcinomas of the breast: 52 tumors were
stage I, 61 were stage II, and 30 were stage III. Tumors were classified as
MRP-1/CD9 positive when a band intensity of >30% compared with
positive control cells, ZR-75-30 were evaluated with the antibody M31-15,
and those with intensities <30% as negative. Moreover, these results were
ascertained by immunostaining. Tumor specimens classified as MRP-1/
CD9 positive using Western blotting had >50% of the cancer cells immunostained with M31-15, and those classified as MRP-1/CD9 reduced
had <50% of the cancer cells immunostained with M31-15. There were 97
patients with MRP-I/CD9 positive tumors and 46 patients whose tumors
had reduced MRP-I/CD9 levels. The disease-free rate of the former group
of patients was strikingly higher than that of the latter (84.7% versus
51.4%, P < 0.001). Similarly, the overall survival rate was also signifi
cantly different between the two groups (93.6% versus 69.6%, P = 0.004).
Multivariate analysis with the Cox regression model indicated that MRP1/CD9 positivity correlated better with disease-free survival (P < 0.001)
than
that
ated
may
Kilaku, Osaka 530
ical grade, and estrogen receptor status. Recently, the focus pertaining
to factors of potential prognostic utility has turned to the expression of
molecular markers. A few of these have been reported to be associated
with poor prognosis, and their presence or levels may be an indication
for adjuvant therapy. For example, the c-erbB-2 oncogene overexpression is found to be associated with shorter survival, particularly in
breast cancer patients with involved lymph nodes (3-5). Mutations of
the tumor suppressor gene f>53 also may be of prognostic importance,
especially for patients with node-negative disease (6. 7). On the other
hand, it has been reported that nm23 protein expression in breast
cancer was associated with good prognosis and absence of lymph
node métastases,but that low nm23 mRNA levels correlated with the
presence of lymph node métastasesat surgery (8, 9).
We have previously generated a murine MAb\ M3I-I5. that in
hibits cell motility and established that the antibody recognizes the
MRP-1.' a transmembrane glycoprotein (10). The amino acid se
quence of MRP-1 is identical to that of CD9, a WBC differentiation
antigen, reported in the same year (11). MRP-1/CD9 is expressed not
only by hematopoietic cells, but also by most established cell lines
derived from solid human tumors. However, its role in nonhematopoietic cells remains unknown. In efforts to elucidate the functions of
MRP-1/CD9 in such cells, we transfected various types of cultured
cells with plasmid constructs containing human MRP-I/CD9 cDNA
(12). These experiments revealed that cell motility and growth were
suppressed in the MRP-l/CD9-expressing
cells. In addition, investi
gations with mouse melanoma BL6 cells and the BALB/c-/i(i//i»
mouse system disclosed that the metastatic potential of all transfor
mants expressing MRP-1/CD9 was lower than that of the parent BL6
cells (12). This set of observations indicated that MRP-1/CD9 regu
estrogen receptor, tumor, and lymph node status. Our data suggest
low MRP-1/CD9 expression by tumors of the breast may be associ
with poor prognosis. It is conceivable that testing for MRP-1/CD9
identify node-negative breast cancer patients who are at high risk for
early disease recurrence.
Introduction
Several randomized clinical trials have shown that adjuvant che
motherapy improves the survival of patients with breast cancer (1-3).
However, because it is not clear which clinical characteristics deter
mine the need for adjuvant chemotherapy, it is very difficult to
distinguish patients for whom surgery alone is adequate from those
who may benefit from both surgery and additional treatment. Hence,
the search for sensitive and reliable prognostic tests is of cardinal
importance since they would be valuable for identifying patients for
whom intensive adjuvant therapy is warranted (3). The prognostic
factors that may help predict recurrences of breast cancer include the
number of involved lymph nodes, the size of the tumor, its histologReceived 12/19/95; accepted 2/6/96.
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 (his fact.
1 Supported in part by Grants-in-Aid from the Ministry of Education in Japan and The
lates cell motility, and that it is a receptor for negative signal ligands.
More recently, to investigate whether the levels of MRP-1/CD9 in
tumor tissues are valuable for predicting the clinical behavior of actual
human cancer, we tested MRP-1/CD9 expression in invasive ductal
carcinomas of the breast with immunohistochemical study and immu
noblotting. Thus, we found that MRP-1/CD9 expression diminished
as the clinical stage of a given breast cancer advanced (13). Besides,
primary tumors in almost half of the cases had higher MRP-1/CD9
protein levels than their respective metastatic lymph nodes. These
findings, obtained with immunohistochemical and immunoblotting
methods, were corroborated by determining MRP-1/CD9 gene expres
sion with a quantitative reverse transcription-PCR-based assay. Gene
overexpression was not observed in any of the samples studied (13).
Moreover, we have applied quantitative reverse transcription-PCR
analysis to determine MRP-I/CD9 gene expression in non-small cell
lung cancer and found that low MRP-1/CD9 gene expression by
tumors of the lung may be associated with poor prognosis ( 14). Based
on these results, in this report we present the findings of the prospec
tive evaluation of MRP-1/CD9 protein expression in tumor tissues
3 The abbreviations used are: MAb. monoclonal antibody: MRP-1. motility-related
protein-1; TM4SF. transmembrane 4 supertamily.
Sagawa Foundation for Promotion of Cancer Research (M. M.).
2 To whom requests for reprints should be addressed.
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MRP-I/CD9 AS A PROGNOSTIC FACTOR OF BREAST CANCER
Table 1 Disease-free sun'ival and overall sumval
rale of 143 patients with breas! cancer according to clinicopathological
characteristics
and MRP-Ì/CD9status
MRP-1/CD9
rateMRP-1
survival
rateMRP-1
survival
statusCharacteristicsAge
-'ci77.659.558.285.284.474.546.771.371.750.033.393.373.
-%)48.155.036.070.875.050.020.080.032.150.0086.752.921.451.4P0.005<0.001•C0.0010.0180.0110.0310.1670.010<0.
+(<90.397.788.8100.096.792.476.2100.089.985.780.0100.093.378.393.6
+(77.993.077.895.090.380.855.692.979.362.580.090.983.766.784.7MRP-1
+
of patients)2620252120161020212315161546Disease-free
(no.53445641612794440g537451597MRP-1
(yr)<50g50Estrogen
at surgery
status—+Nodal
receptor
statusNONIN2Tumor
statusT,T2TT.Pathological
stageStage
IStage
IIStage
IIITotal
no. of patientsMRP-1
from 143 breast cancer patients for whom adequate clinicopatholog
ical data were available.
Immunoblot Analysis. Tissue samples containing only cancer cells were
solubilized with 1% CHAPS. An aliquot of the soluble fraction was subjected
to slab gel electrophoresis, followed by Western blotting and probing with the
anti-MRP-1/CD9 MAb, rabbit antimouse IgG (y chain specific; Cappel,
Malvern, PA) and I25l-labeled protein A (DuPont, Boston, MA); Refs. 16 and
Materials and Methods
Clinical Characteristics of the Patients. This study was carried out on
143 patients with invasive ductal carcinoma of the breast. All of them under
went mastectomy or quadrantectomy at the Department of Thoracic Surgery of
the Kitano Hospital Medical Research Institute (Osaka, Japan) between May
1990 and December 1992. Complete clinical records of all of them were
available, and the histopathological diagnoses were fully documented. The
postsurgical pathological stage of each tumor was classified according to the
TNM system (15). The salient clinical features of the patients are presented in
Table 1. The median postoperative follow-up for the patients was 45.7, (range,
30-61) months. This report includes follow-up data as of July 1, 1995. Ten
patients with intraductal carcinoma and nine with two or more forms of cancer
were not included in the study. In addition, four women with stage IV disease
were also excluded because even if their primary tumors are resected, such
patients usually have cancer cells at the metastatic sites. This would make the
disease-free survival analysis of these patients meaningless in the context of
the present study.
Eleven patients had quadrantectomy followed by immediate radiotherapy,
and the other patients had no radiotherapy before recurrence. Adjuvant sys
temic chemotherapy was given according to estrogen receptor status, meno
pausa! status, and lymph node involvement. Node-positive or premenopausal
patients («= 108) underwent treatment by oral 5-fluorouracil (200 mg/day) for
2 years, and, furthermore, 19 patients with N, disease were treated with six
cycles of cyclophosphamide/Adriamycin.
Sixty-two estrogen receptor-positive
patients were treated with tamoxifen (20 mg/day) for 2 years or before
recurrence. Twenty-two patients (postmenopausal node negative and receptor
negative) did not have any further adjuvant treatment. We detected distant
métastasesin 25 patients during the observation period, and among these 25
patients, 3 patients also had local recurrences. Twelve patients had locore-
17). The total protein content applied to the lanes was adjusted to the same
concentration. Soluble extracts of the established human breast cancer cell line
ZR-75-30 were used as positive controls. Band intensity was evaluated with
densitometry.
Immunohistochemical
Assay. Since the MRP-1/CD9 antigen is not pre
served in formalin-fixed paraffin-embedded tissues, we used frozen sections
for these assays. They were performed using an antimouse IgG ABC kit
(Vector Laboratories, Inc., Burlingame, CA) as described previously (13).
Sections incubated with mouse myeloma SP2 supernatant served as negative
reaction controls.
Statistical Analysis. Overall survival curves and disease-free survival
curves were constructed according to the Kaplan-Meier method (18). Com
parisons were made with the log rank test (19). The SAS statistical package
(SAS Institute, Cary, NC) was used for the Cox proportional hazards model;
five factors (age at surgery, T stage, N stage, estrogen receptor status; and
tumor MRP-1/CD9 status) were studied, and scores were assigned to each
variable for the regression analysis (20). The
MRP-1/CD9 expression and T stage, N stage,
investigated using a frequency table. All P
statistical analysis; a P value of <0.05 was
significance.
association between reduction in
and estrogen receptor status was
values are based on two-tailed
considered to indicate statistical
Results
immediately embedded in optimum cutting temperature compound (Miles,
Kankakee, IL) and frozen at -80°C. Frozen sections were cut on the cryostat
Specimen Classification Based on Western Blotting Results.
MRP-1/CD9 expression in breast tumor tissues was analyzed using
Western blotting with MAb 31-15. The major Mr 25,000 MRP-1/CD9
band is usually accompanied by a minor Mr 28,000 band. The major
band was readily evident in the immunoblots of the soluble fractions
of the primary tumors that were classified as positive in the immunohistochemical assays, as described previously (Fig. 1; Ref. 13). By
contrast, the band intensity was weak or entirely absent in the primary
invasive ductal carcinomas that had reduced immunohistochemically
detectable MRP-1/CD9. To classify the specimens on the basis of
to a thickness of 6 ¿un,mounted on poly-D-lysine-coated slides, and either
used immediately or stored at —¿
80°Cfor no more than 2 weeks. To verify the
Western blotting, the M, 25,000 band obtained with the positive
control cells ZR-75-30 was set at 100%. The distribution of the ratio
presence of cancer ceils, a frozen section from each specimen was stained
immediately with H&E. One-half of a given tumor sample containing only
of each specimen to the control is shown in Fig. 2. There was a clear
difference among them with regard to intensity at the point of 3040%. Patient samples with a band intensity of >30% were classified
gional recurrences. After recurrence, locoregional tumors or lymph nodes were
principally resected and then these patients were treated with radiotherapy.
Patients with distant métastaseswere treated with more effective adjuvant
chemotherapy, including cisplatin and pirarubicin.
Surgical Specimens. One-half of each freshly resected tumor tissue was
cancer cells was used for Western blotting.
1245
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MRP-1/CTO AS A PROGNOSTIC
FACTOR OF BREAST CANCER
levels (Figs. 4 and 5). The disease-free survival rates were 84.7% and
51.4% (P < 0.001), respectively (Table 1). Moreover, tumors with
reduced MRP-1/CD9 expression of N0 and N, disease or with T, and
T2 disease were associated with progressively reduced disease-free
survival (Table 1). The overall survival rate of patients with MRP-1/
CD9-positive tumors was 93.6% (Table 1) and that of the individuals
whose tumors had reduced MRP-1/CD9 was 69.6% (P = 0.004).
Prognostic Value of MRP-1/CD9 Status. The variables used in
the Cox regression analysis are shown in Tables 2 and 3. The esti
mated prognostic value of each variable in relation to disease-free
survival and overall survival among the 143 patients is expressed as a
P value. Four variables (MRP-1/CD9 status, estrogen receptor status,
N stage, and T stage) were found to be significant prognostic factors
of survival. MRP-1/CD9 had the most significant P value for diseasefree survival (P < 0.001), and the second with respect to overall
survival (P = 0.0084). Other variables (body weight, serum albumin,
1 2345678
carcinoembryonic antigen, progesterone receptor status, postoperative
hormonal therapy, and chemotherapy) had no relationship with
MRP-1/CD9 status or its prognostic value (data not shown).
Fig. 1. Immunoblot analysis of MRP- 1/CD9 expression in breast tumors. Lane I, breas!
cancer cell line ZR-75-30 (positive control); Lanes 2, 4, and 6-8. primary invasive ductal
carcinomas with reduced MRP-1/CD9 levels; Limes 3 and 5. primary invasive ductal
carcinomas with high MRP-1/CD9 levels.
Discussion
as MRP-1/CD9 positive, and those with intensities <30% as negative.
Of the 143 primary breast cancers studied, 97 (67.8%) were classified
as MRP-1/CD9 positive. There were 46 cases (32.2%) with negative
MRP-1/CD9 expression.
Immunohistochemical
Detection of MRP-1/CD9 in Breast
Cancer Tissues. In primary breast cancers classified as MRP-1/CD9
positive, expression resembled that of normal glands and benign
fibroadenomas (Fig. 3A) since immunostaining was intense and was
seen uniformly at the cell surface membrane (Fig. 3B). There were 46
cases (32.2%) with reduced MRP-1/CD9 expression (Fig. 3C). Im
munostaining of most of these tumors was heterogeneous, and the
staining pattern along cell junctions was not linear, but granular. In
each instance, MRP-1/CD9 expression in metastatic lymph nodes was
lower than that in the corresponding primary tumor (Fig. 3D).
All immunostained sections were examined by two pathologists
who had no knowledge of the patients' clinical status. At least 200
tumor cells were counted per X40 field. Positive tumor cells were
stained equivalent to normal breast glands and benign fibroadenoma
tumor cells. When >50% of the carcinoma cells in a given specimen
were positively stained, the sample was classified as MRP-1/CD9
positive, and when <50% were stained, the sample was classified as
reduced. Overall, the immunoblotting
results agreed well with
those of the imniunohistochemical
assays, and, in seven cases of
discrepancy, the results of Western blotting were used in specimen
classification.
Relationship between Tumor MRP-1/CD9 Immunoreactivity
and Known Prognostic Factors. Analysis of the results obtained
with the 143 breast carcinomas tested revealed no statistically signif
icant relationship (x2) between MRP-1/CD9 expression and the pa
tients' age at surgery or estrogen and progesterone receptor status,
Prevention of metastasis is among the most important problems in
the treatment of patients with malignant tumors (21. 22). Despite
various types of antitumor defense mechanisms, the malignant tumor
cells that invade tissue boundaries and move through the host's
cellular and extracellular matrix barriers resist the shear stresses
arising in the vascular bed, the frictional forces arising between their
peripheries, and the vessel walls. In addition, they traverse capillaries
that are generally rigid and whose diameter is smaller than the tumor
cells. It has been shown that motility of cancer cells is one of the
essential cellular functions closely related to the metastatic process
(21, 22).
We have previously demonstrated that motility of M AC 10 cells,
derived from a human adenocarcinoma of the lung, is inhibited by
MAb M31-15, and that this antibody recognizes MRP-1, a transmem
brane glycoprotein (10). We also determined that the sequence of
MRP-1 is identical to that of CD9. the sequence of which was reported
in the same year by another laboratory (11). Because disruption of cell
to cell contacts induces tumor cell invasion and métastases,it is of
interest in the present context that MAbs to CD9 elicit an enhance
ment of Fc-independent heterotypic adhesion of pre-B cell lines to
bone marrow stromal fibroblasts, but not to bone marrow stroma (23).
Moreover, these antibodies specifically induce homotypic adhesion of
pre-B lymphocytes and augment neutrophil adherence to the endothe-
(N)
30-1
s
u
20-
o.
i—
o
IM
tumor size, or postoperative hormonal therapy and chemotherapy.
However, MRP-1/CD9 expression was associated with lymph node
status (P = 0.045). Thus, whereas 24.7% of the patients with N0 stage
disease had reduced expression, low MRP-1/CD9 was found in 37.2%
of the N, stage patients and 52.6% of those with N2 stage.
Association of Tumor MRP-1/CD9 Status with Disease-free and
Overall Survival. Comparing survival among the 143 patients re
vealed that the disease-free and overall survival rates for the 97
patients with MRP-l/CD9-positive tumors differed significantly from
those of the 46 individuals whose tumors had reduced MRP-I/CD9
25-
15-
o
.o
g
5-
-ñUil
0-10
10-20 20-30 30-40 40-50 50-60 60-70 70-80 80-9090-100
100<
r;.)
Ratio of western blotting value
Fig. 2. Distribution of the ratio of Western blotting values of tumor specimens:the
control cell line ZR 75-30.
1246
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MRP-1/CD9
AS A PROGNOSTIC
FACTOR OF BREAST CANCER
B
•¿
•¿
.
~
•¿
4 .
V
Fig. 3. Immunohistochemical staining of human breast tissues with the MRP-l/CD9-specific
MAb M31-15. Avidin-biotin-peroxidase complex procedure. A, the cell surface in a
benign fibroadenoma was strongly stained, ß.positive immunostaining of an invasive ductal carcinoma of the breast. C. invasive ductal carcinoma with reduced MRP-1/CD9 expression.
D. metaslatic lymph node corresponding to the invasive carcinoma shown in B. Note that staining intensity is lower than in the primary tumor. A-D, X 150.
S
100-ça
-75
c¿ go
80-b
1^~~V,
around 15 members that are variously expressed by leukocytes and a
variety of mammalian tissues, as well as on the surface of two types
of parasites (26). Although their precise functions are still unknown,
current data largely suggest that TM4SF proteins are involved in the
regulation of cell development, proliferation, activation, and motility.
The melanoma-associated antigen ME491/CD63 is an interesting
member of this superfamily. It is preferentially expressed in early
stages of tumor progression, but expression declines significantly and
sometimes disappears entirely in advanced, rapidly growing melano
mas and metastatic melanoma cells (27). In addition, the predicted
~1"1
(n=97)v_1MRP-1/CD9MRP-1/CD9
positive
60-W
50-<B
40-^
30-S
=46)P<0.0010
reduced (n
20-I
10-f*S
O "*"T-^1
10
20
30
40
5060
7
\7v)100-£
97)~^~
Months after Surgery
90-(2
Fig. 4. Disease-free survival in 143 patients with invasive ductal carcinoma of the
breast ¡nrelation to the tumor MRP-I/CD9 status (P < 0.001). P value was determined
with the log rank test.
positive
(n =
^i_^MRP-1/CD9
80-n|60-i
(n=46)P
reduced
Hum probably caused by an adhesion-promoting activation event
within the endothelial cells (24). These findings would suggest a
direct participation of CD9 in signal transduction. Since the MAb to
MRP- 1/CD9 inhibits the motility of various types of cancer cells, it is
conceivable that the glycoprotein affects motility and invasiveness by
modulating the levels of cellular adhesion (10, 13).
MRP-1/CD9 belongs to the TM4SF, which generally has four
highly conserved hydrophobic domains that are assumed to span the
lipid bilayer of the cell membrane (25, 26). The TM4SF consists of
50-w
40-75
30-£20O
= 0.004
10n -MRP-1/CD9
10
20
30
40
50
60
70
Months after Surgery
Fig. 5. Overall survival of the 143 patients with invasive ductal carcinoma of the breast
in relation to the tumor MRP-I/CD9 status (P = 0.004).
1247
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MRP-I/CD9 AS A PROGNOSTIC FACTOR OF BREAST CANCER
Table 2 Value of five variables in predicting disease-free survival of 143 patients with breast cancer
VariablesMRP-1/CD9-+Age
interval-1.4769
ß
score010101123401295%
SE
Wald's X2
Hazard ratio
0.109-0.4790.1456 0.3781
15.2535
0.228
0.560-2.390-1.6012 0.3703
0.1546
1.157
0.086-0.4710.6842 0.433
13.6761
0.202
1.348-2.9160.7155 0.1969
12.0755
1.982
10.7729
2.045
Confidence
(yr)<50g50Estrogen
receptor-+Tumor
statusT,T2TjT4Nodal
statusN„N,N2Assigned
0.218
1.334-3.136P<0.0010.69420.00020.00050.0010
Table 3 Value of five variables in predicting overall xunival of 143 patients with breast cancer
VariablesMRP-1/CD9-+Age
(yr)<50SSOEstrogen
x2-1.4083
score0101011234012ß
SE
Wald's
interval0.245
ratio
6.9566-0.1422
0.5339
0.086-0.6960.867
0.0699-1.3385
0.5381
0.302-2.4900.262
4.28870.8360
0.6463
0.074-0.9312.307
9.02120.7262
0.2784
1.337-3.9812.067
Confidence
receptor-+Tumor
statusTIT,T3T4Nodal
statusNON,N2Assigned
0.3153
5.303795%Hazard
product encoded by KAII, a recently described metastasis suppressor
gene of prostate cancer, is identical to R2 and C33, which also belong to
the TM4SF (28-30). It is thought that decreased KAI1 expression may be
involved in the malignant progression of prostate cancer (28). These
observations on other systems are very consistent with our data that the
level of MRP-1/CD9 expression is inversely related to the clinical stage
of a given cancer of the breast. Thus, MRP-l/CD9-positive tumors were
evident in 73% of the patients whose tumors were in stages I or II, but
only in 50% of patients with stage III tumors.
As the present results indicate, reduction in MRP-1/CD9 expression
by primary invasive ductal carcinomas of the breast was associated
with poor prognosis. The link between diminished MRP-1/CD9 and
poor prognosis was independent of tumor stage, lymph node status,
and estrogen receptor status. Reduction in tumor MRP-1/CD9 expres
sion was strongly associated with an increased risk of recurrence
among the patients with N0 and N, stage disease and with T, and T2
tumors. However, the interpretation of these findings has to be made
with caution in view of the relatively short postoperative follow-up
(30-61 months) of the patients.
Adjuvant endocrine therapy and chemotherapy have signifi
cantly improved the disease-free survival of patients with early
stage breast cancer (1-3). However, the role of adjuvant chemo
therapy after mastectomy, particularly in women whose disease is
confined to the breast, has remained unresolved. It is possible that
patients with MRP-l/CD9-positive
tumors may not require adju
vant chemotherapy after mastectomy. By contrast, patients whose
tumors have reduced MRP-1/CD9 levels, even if confined to the
1.114-3.835P0.00840.79150.03840.00270.0213
breast, have a poor prognosis, and may therefore benefit from
adjuvant treatment. It is conceivable that testing tumors for MRP1/CD9 status, in combination with other molecular and biochem
ical assays, may improve the prognostic evaluation of breast cancer
patients and enhance the clinician's ability to prospectively iden
tify patients who will have early disease recurrence
require adjuvant chemotherapy.
and who will
Acknowledgments
We thank Dr. Masato Vagita for critical editorial help with the manuscript,
Tatsuya Hirai for the technical assistance, Drs. Shinji Sawada and Tadashi
Oobayashi for the histopathological examination of the tumor samples, and Dr.
Isao Horiguchi for statistical analysis.
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1249
Downloaded from cancerres.aacrjournals.org on June 11, 2017. © 1996 American Association for Cancer Research.
Motility-related Protein-1 (MRP-1/CD9) Reduction as a Factor of
Poor Prognosis in Breast Cancer
Masayuki Miyake, Keiji Nakano, Shin-ichi Itoi, et al.
Cancer Res 1996;56:1244-1249.
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