СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes
Aim: To determine frequency of tumors with immunohistochemical markers of cancer stem cells (CSC) CD44+/CD24− in patients with breast cancer (BC) of different molecular subtype and to evaluate their prognostic value. Object: Surgical material of 132 patients with BC stage I–II, age from 23 to 75 yea...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2015
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| Cite this: | СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes / S.V. Chekhun, T.V. Zadvorny, Yu.O. Tymovska, M.F. Anikusko, O.E. Novak, L.Z. Polishchuk // Experimental Oncology. — 2015. — Т. 37, № 1. — С. 58-63. — Бібліогр.: 38 назв. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1860044795857076224 |
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| author | Chekhun, S.V. Zadvorny, T.V. Tymovska, Yu.O. Anikusko, M.F. Novak, O.E. Polishchuk, L.Z. |
| author_facet | Chekhun, S.V. Zadvorny, T.V. Tymovska, Yu.O. Anikusko, M.F. Novak, O.E. Polishchuk, L.Z. |
| citation_txt | СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes / S.V. Chekhun, T.V. Zadvorny, Yu.O. Tymovska, M.F. Anikusko, O.E. Novak, L.Z. Polishchuk // Experimental Oncology. — 2015. — Т. 37, № 1. — С. 58-63. — Бібліогр.: 38 назв. — англ. |
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| description | Aim: To determine frequency of tumors with immunohistochemical markers of cancer stem cells (CSC) CD44+/CD24− in patients with breast cancer (BC) of different molecular subtype and to evaluate their prognostic value. Object: Surgical material of 132 patients with BC stage I–II, age from 23 to 75 years, mean age — 50.2 ± 3.1 years was studied. Methods: Clinical, immunohistochemical (expression CD44+/CD24−), morphological, statistical. Results: BC is characterized by heterogeneity of molecular subtypes and expression of markers (CD44+/CD24−). Immunohistochemical study of expression of CSC markers in surgical material has detected their expression in 34 (25.4%) patients with BC of different molecular subtypes. The highest frequency of cells with expression of CSC marker was observed in patients with basal molecular subtype (44.8% patients). Most of BC patients with phenotype CD44+/CD24 had stage I of tumor process (34.3%). Statistical processing of data has showen that Yule colligation coefficient equaled 0.28 (р > 0.05) that argues poor correlation between stage of tumor process and number of tumors with positive expression of CSC markers. Statistical processing of data has showen high correlation between presence of cells with expression of CSC markers and metastases of BC in regional lymph nodes (Yule colligation coefficient equals 0.943; р < 0.5). Difference in overall survival of patients with BC of basal molecular subtype depending on expression of CSC CD44+/CD24− markers was detected. Survival of patients with basal BC was reliably higher at lack in tumors of cells with CSC markers CD44+/CD24− and, correspondingly, lower at presence of such cells (р < 0.05). In patients with BC of luminal (A and B), HER-2-positive subtypes, significant change in survival of patients depending on expression of CSC markers was not determined (р > 0.05). Conclusion: Significance of tumor cells with markers CD44+/CD24− within the limits of molecular subtype of BC may be additional criterion for advanced biological characteristic of BC, and in patients with BC of basal molecular subtype — for predictive evaluation of individual potential of tumor to aggressive clinical course. Key Words: breast cancer, cancer stem cells, СD44+/CD24−, molecular subtype.
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58 Experimental Oncology 37, 58–63, 2015 (March)
СD44+/CD24− MARKERS OF CANCER STEM CELLS IN PATIENTS
WITH BREAST CANCER OF DIFFERENT MOLECULAR SUBTYPES
S.V. Chekhun1, *, T.V. Zadvorny1, Yu.O. Tymovska2, M.F. Anikusko2, O.E. Novak2, L.Z. Polishchuk1
1R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine,
Kyiv 03022, Ukraine
2Kyiv City Clinical Oncological Center, Kyiv 03022, Ukraine
Aim: To determine frequency of tumors with immunohistochemical markers of cancer stem cells (CSC) CD44+/CD24− in patients
with breast cancer (BC) of different molecular subtype and to evaluate their prognostic value. Object: Surgical material of 132 pa-
tients with BC stage I–II, age from 23 to 75 years, mean age — 50.2 ± 3.1 years was studied. Methods: Clinical, immunohisto-
chemical (expression CD44+/CD24−), morphological, statistical. Results: BC is characterized by heterogeneity of molecular
subtypes and expression of markers (CD44+/CD24−). Immunohistochemical study of expression of CSC markers in surgical
material has detected their expression in 34 (25.4%) patients with BC of different molecular subtypes. The highest frequency of cells
with expression of CSC marker was observed in patients with basal molecular subtype (44.8% patients). Most of BC patients with
phenotype CD44+/CD24 had stage I of tumor process (34.3%). Statistical processing of data has showen that Yule colligation
coefficient equaled 0.28 (р > 0.05) that argues poor correlation between stage of tumor process and number of tumors with positive
expression of CSC markers. Statistical processing of data has showen high correlation between presence of cells with expression
of CSC markers and metastases of BC in regional lymph nodes (Yule colligation coefficient equals 0.943; р < 0.5). Difference
in overall survival of patients with BC of basal molecular subtype depending on expression of CSC CD44+/CD24− markers was
detected. Survival of patients with basal BC was reliably higher at lack in tumors of cells with CSC markers CD44+/CD24− and,
correspondingly, lower at presence of such cells (р < 0.05). In patients with BC of luminal (A and B), HER-2-positive subtypes,
significant change in survival of patients depending on expression of CSC markers was not determined (р > 0.05). Conclusion:
Significance of tumor cells with markers CD44+/CD24− within the limits of molecular subtype of BC may be additional criterion
for advanced biological characteristic of BC, and in patients with BC of basal molecular subtype — for predictive evaluation of in-
dividual potential of tumor to aggressive clinical course.
Key Words: breast cancer, cancer stem cells, СD44+/CD24−, molecular subtype.
Breast cancer (BC) is one of the most widespread
forms of female cancer pathology in many countries
of the world. Dangerous situation concerning BC is ob-
served in Ukraine as well: according to updated infor-
mation of National Cancer Register of Ukraine (2012)
BC morbidity and mortality (raw indices) are respec-
tively 72.5 and 32.2 per 100 000 of female population.
Only 13.3% patients has received surgical treatment
alone, while combined or complex treatment —
68.5% [1] that evidences that the significant number
of patients had advanced stages of BC. Treatment
results, being not always satisfactory due to tumors’
chemo- and radioresistances and significant variability
of BC clinical course [2–6], serve the basis for more
pronounced study of BC bio logy from positions of its
origin, growth and metastasis.
Intense molecular-biological studies have deter-
mined that BC is complicated multi-stage process
of accumulation of structural and functional effects
of genes, which are associated with proliferation, neo-
angiogenesis, intercellular adhesion disturbances, pe-
culiarities of microenvironment of tumor cells and their
clonal evolution. The main result of these studies was
the identification of BC as heterogeneous disease with
different properties of growth and metastasis [7, 8].
This is confirmed by identification of different BC mo-
lecular subtypes depending on expression of estrogen
receptors (ER), progesterone receptors (PR) and
human epidermal growth factor type 2 (HER2/neu).
Basing on this, the following BC subtypes were de-
termined, namely: luminal A, luminal B, basal or triple-
negative, HER2/neu-positive [9–12]. Mammologists
are guided by these biological subtypes while treat-
ment process and evaluation of disease prognosis.
Despite widespread use of such classification
in clinical practice, it is not always possible to predict
precisely the clinical course of disease and tumor
“response” to the treatment. It is due to the fact that
BC of one histological type or even of one molecular
subtype is characterized by different proliferative
potential of tumor cells, adhesion disturbances, pecu-
liarities of tumor microenvironment, which altogether
determine not only intercellular, but also intracellular
biological heterogeneity [13–15]. Literature data con-
firm the dependence of unfavorable BC clinical course
and prognosis from tumor biological heterogeneity,
genetic and proteomic changes in tumor cells [16–18].
In this aspect, cancer stem cells (CSC) attract atten-
tion. According to the conception of CSC significance
for tumor occurrence and growth, such cells possess
specific features. Like normal stem cells, they have
capability to grow, can self-renew and differentiate into
various types of cells. Their specific feature is: while CSC
division one of the daughter cells preserves properties
Submitted: January 05, 2015
*Corresponding: E-mail: chekhun@yahoo.com
Abbreviations used: BC — breast cancer; CSC — cancer stem
cells; ER — estrogen receptors; HER2/neu — human epidermal
growth factor type 2; PR — progesterone receptors.
Exp Oncol 2015
37, 1, 58–63
Experimental Oncology 37, 58–63, 2015 (March) 59
of parent cell, while cells proliferation is associated with
CD44+/CD24− phenotype [19–21]. Cells with such
phenotype are called tumorogenic or tumor-initiating
cells due to the features of their proliferative potential.
Studies on CSC have determined that biological and
molecular heterogeneity of BC depends on the amount
of such cells [22], their microenvironment and expression
of certain cytokines [23]. Biological properties of these
cells were studied on experimental cells lines in vitro [24].
Earlier in our studies on CSC in cell lines of BC (T47D,
MCF-7, MDA-MB 231, MDA-MB 468) we have deter-
mined some regularities of CSC markers expression
depending on their molecular phenotype [25].
Thus, in modern literature, both the significance
of BC molecular phenotypes for characterization
of BC heterogeneity and the hypothesis of intratumoral
heterogeneity formation, in which CSC may play sig-
nificant role, are actively discussed. But CSC associa-
tion with clinical features of BC of different histological
types, with metastasis and patients survival remains
unclear. For this reason, the study of BC heterogeneity
and cellular peculiarities of tumors using immunohis-
tochemical markers of CSC is relevant problem, which
solution will promote extension of knowledge on bio-
logy of tumor growth in mammary gland and improve
patients’ treatment.
Taking into account above-stated, the aim of this
study was to determine the prevalence of tumors with
immunohistochemical markers of cancer stem cell
CD44+/CD24− in patients with BC of different molecu-
lar subtype and evaluate their prognostic significance.
MATERIALS AND METHODS
Our studies are based on retrospective analysis
of results of examination, treatment and survival
of 132 BC patients with I–II stages, who received
special treatment in Kyiv City Clinical Cancer Center
during 2005–2007. The samples of surgical mate-
rial of BC were studied. These samples were stored
in clinical database of the Department of Antitumor
Therapy Mechanisms of R.E. Kavetsky Institute of Ex-
perimental Pathology, Oncology and Radiobiology,
NAS of Ukraine. The tumor stage was determined
according to the International Tumor Classification
(TNM, Edition 6, 2002). Histological type of tumors
was verified on histological sections of tumors’ paraffin
blocks (staining with hematoxylin and eosin) according
with the WHO International Histological Classification
(2006). Depending on clinical indications, patients
underwent organ-saving surgeries or radical Mad-
den mastectomies, and adjuvant polychemotherapy
(CAF or AC schemes with 21 day interval, number
of courses varied from 4 to 6), according with the
approved in Ukraine Standards of BC Patients’ Treat-
ment. Postoperative radiation therapy was performed
on postoperative scar, axillary, parasternal and supra-
clavicular regions; the single focal dose was 2 Gy, and
the total focal dose — 40 Gy.
For evaluation of BC molecular subtype we per-
formed the immunohistochemical studies of expres-
sion of three molecular markers that are widely used
nowadays for immunohistochemical BC diagnosis: ER,
PR, Her2/neu. We used histological sections (4 micron
thickness) of paraffin blocks. The following monoclonal
antibodies were used: antiER — clone 1D5, antiPR-
clone PgR636, antiHER/2neu — clone c-erbB-2 (Dako
Cytomation, Denmark). The number of cells with posi-
tive markers expression were determined taking into
account the level of immunohistochemical reaction:
“+++” — strong, “++” — moderate, “+” — low, “0” —
lack of expression. On the parallel paraffin sections the
immunohistochemical study of CD24 and CD44 expres-
sion in tumor cells were carried out. As primary antibo-
dies, monoclonal antibodies specific to CD24 (clone
SN3b, ThermoScientific, USA) and to CD44 (CD44/
HCAM, clone 156–3C11, Diagnostic BioSystems, USA)
were used in dilutions according to the manufacturer
guideline. The number of immunopositive tumor cells
more than 10% was taken as positive marker expres-
sion. For visualization of the results of all immunohisto-
chemical reactions EnVision system (Dako LSAB2 sys-
tem, Denmark) was used according to the manufac-
turer guidelines; histological sections were stained with
Mayer’s hematoxylin. Results of immunohistochemical
reactions were analyzed using optic microscope XSP-
137-BP, JNOEC, magnification × 200–400.
Statistic processing of the obtained results was car-
ried out using methods of variation statistics by program
Statistica v. 6.0. For evaluation of correlation between
expression of studied markers and clinical and patho-
logical features of BC, we used the Pearson coefficient
of mutual conjugation (c), Yule coefficient of associa-
tion, Cox regression analysis. Evaluation of survival was
carried out using Kaplan — Meier method starting from
the date of treatment beginning, using log-rank test.
Differences were considered significant at p ≤ 0.05.
RESULTS AND DISCUSSION
General clinical description of 132 BC patients
with stage I–II is presented in the Table 1. The number
of BC patients with stage I was 26.5%, with stage II —
73.5%. Patients’ age was varied from 23 to 75 years,
mean age was 50.2 ± 3.1 years. The majority of pa-
tients were in age interval 41–70 (Fig. 1, Table 1).
0
10
20
30
40
50
60
≤30 30–40 41–50 51–70 71–75
Age, years
BC
p
at
ie
nt
s,
%
Fig. 1. Distribution of BC patients with stage I–II by age
60 Experimental Oncology 37, 58–63, 2015 (March)
Table 1. General clinical description of BC patients with stage I–II
Index Number of patients
n %
Total number of patients 132 100
Patients age (years)
Mean 50.2 ± 3.1
Age variation 23–75
Menstrual function
Preserved 47 35.6
Menopause 85 64.4
BC stage (after TNM)
Stage І 35 26.5
Stage II 97 73.5
Metastases in regional lymph nodes (category N)
N0 95 72.0
N1–3 37 28.0
Distant metastases (category М)
М0 132 100.0
Morphology of BC
Infiltrative ductal carcinoma 92 69.7
Infiltrative lobular carcinoma 40 30.3
Differentiation grade
G1(high) 38 28.8
G2 (moderate) 64 48.5
G3 (low) 30 22.7
Molecular subtype
Luminal А: ER+PR+HER2/neu− 54 40.9
Luminal B: ER+PR+HER2/neu+ 32 24.2
Basal (triple negative): ER−PR−HER2/neu− 29 22.0
Hеr2/neu-positive:
ER−PR−HER2/neu+ 17 12.9
According to the results of complex examination
of patients (X-ray, ultrasound, etc.), metastases in re-
gional lymph nodes (N1–3) were detected in 28.0%
cases, distant metastases were not detected. Mor-
phological study has determined infiltrating ductal BC
(69.7%) more often, than lobular BC (30.3%). More
often moderate differentiation of BC (48.5%) as com-
pared with high and low differentiation (28.8 and
22.7%, correspondingly) was observed.
Analysis of the results of immunohistochemical
study of ER, PR and Her2/neu expressions evidenced
luminal A subtype in 40.9% cases, luminal B subtype —
in 24.2%, basal subtype — in 22.0%, and Her2/neu-
positive subtype — in 12.9% cases (Table 1).
More detailed clinical and pathomorphological
description of tumor process in BC patients depend-
ing on molecular subtype of BC is shown in Table 2.
Distribution of patients by mean age has not detected
significant difference (p > 0.05). Most patients with
luminal A, basal and Her2/neu-positive molecular sub-
types (70.4–72.4%) were menopausal, while among
patients with luminal B subtype such patients were
43.8%. In all molecular subtypes, mostly stage II was
diagnosed, especially in patients with luminal A (83.4%)
and basal (82.8%) subtypes, while in patients with
Her2/neu-positive and luminal B subtypes, stage II was
in 53.0 і 59.4% cases, respectively. Frequency of me-
tastasis in regional lymph nodes was significantly lower
in BC patients with both luminal subtypes.
In patients with basal and Her2/neu-positive sub-
types the number of patients without metastases and
with metastases was almost the same: basal subtype
(51.7 and 48.3%, respectively) and Her2/neu-positive
subtype (52.9 and 47.1%, respectively). Irrespectively
of tumor molecular subtype, ductal carcinoma prevailed
in all patients, varying from 55.5% in luminal A subtype
to 79.3% in basal (triple negative) subtype.
Table 2. Distribution of BC patients with different molecular subtypes ac-
cording to clinical data
Indices
Molecular subtype
Luminal А
n = 54
(100%)
Luminal B
n = 32
(100%)
Basal
n = 29
(100%)
Hеr2/neu-
positive
n = 17
(100%)
n % n % n % n %
Mean age, years 51.3 ± 4.7 46.8 ± 2.5 48.1 ± 1.8 50.6 ± 4.1
Reproductive period, n = 47
Menopausal period, n = 85
16
38
29.6
70.4
18
14
56.2
43.8
8
21
27.6
72.4
5
12
29.4
70.6
Stage
Stage І, n = 35
Stage ІІ, n = 97
9
45
16.6
83.4
13
19
40.6
59.4
5
24
17.2
82.8
8
9
47.0
53.0
Metastases in lymph nodes
No metastases, N0, n = 95 45 83.3 26 81.2 15 51.7 9 52.9
Metastases in regional
lymph nodes, N1–3, n = 37 9 16.7 6 18.8 14 48.3 8 47.1
Histological subtypes
Infiltrative ductal carcino-
ma, n = 92 30 55.5 25 78.1 23 79.3 14 82.3
Infiltrative lobular carcino-
ma, n = 40 24 44.5 7 11.9 6 20.7 3 17.7
Differentiation grade
High, n = 38 18 33.3 10 31.3 7 24.1 3 17.6
Moderate, n = 64 22 40.7 9 28.1 21 72.4 12 70.6
Low, n = 30 14 26.0 13 40.6 1 3.5 2 11.8
The significant priority of moderate grade of tumor
differentiation was determined in luminal A (40.7%),
basal (72.4%) and Her2/neu-positive (70.6%) subtypes,
and only in luminal B subtype the low grade of tumor dif-
ferentiation (40.6%) prevailed. These data evidenced the
intertumoral heterogeneity of BC both by clinical indices
and by pathomorphological tumor features.
Immunohistochemical study of CSC markers ex-
pression in surgical material detected their expression
in 34 (25.4%) BC patients, who had different molecu-
lar subtypes. Distribution of this marker expression
within each molecular subtype (Table 3) showed that
frequency of expression varied: the highest num-
ber of tumors with of CSC markers expression was
among the patients with luminal B and basal subtypes
(28.1 and 44.8%, respectively), while in patients with
luminal A and Her2/neu-positive subtypes it was
lower (16.6 and 17.6%, respectively). The highest
frequency of cells with CSC marker expression was
revealed in BC patients with basal molecular subtype.
The prevalence of cells with CSC markers depend-
ing on cells’ molecular phenotype was confirmed
by Pearson coefficient mutual conjugation (c = 0.54).
This indicates the existing of moderate correlation
of frequency of cells with CSC markers with tumor
molecular subtype (р < 0.05).
Table 3. Prevalence of tumors with CSC markers expression in BC pa-
tients with different molecular subtypes
Molecular subtype of BC
BC patients with ex-
pression of CSC
marker
CD44+/CD24−
n %
Luminal А: ER+PR+Hеr2/neu−, n = 54 (100%) 9 16.6
Luminal B: ER+PR+Hеr2/neu+, n = 32 (100%) 9 28.1
Basal: ER−PR−Hеr2/neu−, n = 29 (100%) 13 44.8
Hеr2/neu-positive: ER−PR−Hеr2/neu+, n = 17 (100%) 3 17.6
Among criteria, which have essential clinical sig-
nificance for prognosis evaluation and conduction
adequate therapy of BC patients, the stage of tumor
process is crucial.
Experimental Oncology 37, 58–63, 2015 (March) 61
The majority of BC patients with CD44+/CD24−
phenotype had BC I stage (34.3%). Statistic analy-
sis showed that Yule association coefficient was
0.28 (р > 0.05), which underlines poor correlation
between tumor process stage and number of tumors
with positive CSC markers expression (Table 4).
Table 4. Distribution of tumors with CSC marker CD44+/CD24− expres-
sion in BC patients with I–II stage
Characteristic
of patients
Tumors with expression of CSC marker CD44+/CD24-
n %
Stage І, n = 35 12 34.3
Stage ІІ, n = 97 22 22.7
Taking into account literature data on pos-
sible impact of CSC expression in primary tumor and
higher proliferative, invasive and metastatic potential
of tumors, it was of certain interest to study the cor-
relation between frequency of tumors with CD44+/
CD24− markers in patients with or without regional
metastases. Among total number of patients with-
out metastases the CD44+/CD24− markers was
observed in 7 (7.4%) cases, while among those with
metastases such phenotype was detected more of-
ten — in 27 (72.9%) cases (Table 5). Statistic analysis
showed high correlation between the presence of cells
with CSC markers expression and BC metastases
in regional lymph nodes (Yule association coefficient:
0.943; р < 0.05).
Table 5. Distribution of tumors with CSC marker CD44+/CD24− expres-
sion in BC patients without metastases and with regional metastases
Characteristic of patients
Tumors with expression of CSC
marker CD44+/CD24−
n %
No metastases, (N0), n = 95 (100%) 7 7.4
Metastases in regional lymph nodes
(N1–3), n = 37 (100%) 27 72.9*
Notes: *р < 0.05 compared to group of patients without metastasis.
In clinical studies the significance of BC differen-
tiation grade for clinical course of disease was evi-
denced. For this reason, further analysis of obtained
results of immunohistochemical study was devoted
to comparison of CD44+/CD24- expression in BC with
different tumor differentiation (Table 6). The analysis
of distribution of tumors with phenotype CD44+/
CD24− showed that in group of patients with high and
moderate tumor differentiation the amount of such
tumors was lower (18.4 and 25.0%, respectively) than
in patients with low differentiation (36.7%). Statistic
processing detected poor correlation between tumor
differentiation grade and expression of CSC markers
(Pearson coefficient c = 0.3; р > 0.05).
Table 6. Distribution of tumors with CSC marker CD44+/CD24- expres-
sion in BC of different differentiation grade
BC differentiation grade Number of tumors with phenotype CD44+/CD24−
n %
High, n = 38 (100%) 7 18.4
Moderate, n = 64 (100%) 16 25.0
Low, n = 30 (100%) 11 36.7
Taking into account obtained results, which are
characterized by variability of molecular subtypes
in BC patients and CSC markers expression, it was
important to evaluate significance of the latter as cri-
teria of patients’ overall survival. Fig. 2 shows curves
of overall survival of BC patients with different molecu-
lar subtypes depending on presence or lack of tumor
cells with CSC markers. As it could be seen from
presented data, survival of patients with basal BC was
significantly higher at lack of the cells with phenotype
CD44+/CD24− in tumors and, correspondingly, lower
at presence of such cells (р < 0.05). In patients with
BC of luminal (A and B) and HER-2-positive subtypes,
significant changes in survival depending on CSC
markers expression were not found (р > 0.05).
Basal subtypes
0
20
40
60
80
100
0 10 20 30 40 50 60
Months
Su
rv
iva
l,
%
CSC markers –
CSC markers +
Log rank test, p<0.05
Fig. 2. Overall survival of BC patients with different molecular sub-
types depending on presence or lack of SCS markers in tumor cells
In order to determine the significance of using
CSC markers CD44+/CD24− in clinical practice, Cox
regression analysis was carried out. Its results suggest
the possibility of using this index (phenotype of CSC
CD44+/CD24−) as marker for prediction of individual
prognosis for patients with basal BC (Table 7).
Table 7. Results of Cox regression analysis
Marker Molecular subtype β p
CD44+/CD24− Luminal А −0.11 > 0.05
Luminal B −0.21 > 0.05
HER-2/neu-positive 0.17 > 0.05
Basal (triple negative) 0.57 < 0.05
Thus, obtained results show that BC is character-
ized by heterogeneity by not only tumors’ morphology,
but also by molecular subtypes and by CSC markers
expression (CD44+/CD24−). Genetic heterogeneity
of invasive forms of BC is reflected in wide range of his-
tological types, various tumor differentiation grade and
variability of disease clinical course. As it is known, the
luminal subtype A is referred to the low-aggressive
tumors with favorable prognosis; on the contrary, the
luminal B subtype is characterized by aggressiveness
and worse prognosis. Other subtypes — HER2-positive
and triple negative (basal subtype) — are referred
to the tumors with worse prognosis of survival, though
in the first one the HER2/neu expression occurs, while
in the latter no changes of this cancer oncogene were
detected. At the same time, exactly the basal sub-
type is characterized by the worst patients’ survival
rates. It can be suggested that in aggressive course
of BC of basal subtype the particular role is played
by cells with CSC phenotype CD44+/CD24−. In the
relationship “CSC and cancer” many unsolved ques-
tions remain. For this reason, CSC origin, their evolu-
tion, role of such molecular markers as CD44 and
62 Experimental Oncology 37, 58–63, 2015 (March)
CD24 in different stages of tumor genesis are under
wide discussion [12, 26–28].
Population of CSC is not large as compared with
general population of cells in tumor, but it can seri-
ously impact biological properties of BC. The fol-
lowing literature data confirm this. In BC patients the
connection between expression of cells with CD44+/
CD24− and aggressive clinical course and metastasis
was determined [29, 30]. The disseminated cells with
CD44+/CD24− markers appear already at the early
stage of tumor genesis in mammary gland [31].
The correlation between CSC CD44+/CD24− and the
epidermal growth factor receptor (EGFR) in invasive
variant of ductal carcinoma, which can favor higher
proliferation of tumor cells, was found. Increase of pro-
portion of cells with phenotype CD44+/CD24−/low
is significant for manifestations of BC aggressiveness
and increased metastasis risk [32]. There are signs
that tumor cells with mentioned phenotype play cer-
tain role not only in activation of metastasis, but also
in formation of resistance to cytostatic drugs [33].
Subpopulations of cells with CSC phenotype are rela-
tively more radioresistant, so it can be associated with
development of relapses after radiotherapy [34]. One
more important fact is the lack of correlation between
cells with phenotype CD44+/CD24− and response
to hormone therapy in patients with receptor-positive
forms of BC [35].
At the same time, it was determined that fraction
of tumor cells CD44+/CD24−/low is quite flexible and
depends on many factors, including signals of micro-
environment. For example, under hypoxia of tumors
in cancer patients with CSC phenotype CD44+/CD24−/
low, the prognosis is significantly worse. Some authors
noticed the changes of CD44+ and CD24- expressions
in metastases as compared with primary tumor [36].
We also confirmed this fact in immunohistochemi-
cal study of CD44s expression in primary tumors
and implanted metastases of serous ovary cancer.
It was found that changes of this marker expression
in implanted metastases had different tendencies: only
in 26.7% of patients CD44s expression did not change,
while the number of CD44s-positive cells increased
in 46.6% cases and in 26.7% cases decreased [37].
These changes underline the individual nature of in-
tercellular adhesion molecule expression in implanted
metastases that can be the evidence of clonal nature
of metastasis or changes on the level of tumor-host
(metabolic, hormonal etc.). If we take into account
the study of M. Shi pitsin et al. [38], the relations be-
tween cells with different phenotype in tumor popula-
tion are depended on their differentiation response
to the activation of TGF-β signal pathway.
Obtained results demonstrate multiple functions
of CSC and their connection with unequal manifes-
tations of BC aggressiveness of different molecular
subtypes. Undoubtedly, the role of CSC in BC, as well
as in tumors of other genesis, their association with
patients’ hormonal status, peculiarities of microenviron-
ment and other important functions of these cells are
not fully identified. At the same time, despite existing
disputable issues, CSC may be considered as very
interesting for active future studies of the role of these
cells at different stages of tumor genesis. It is important
for understanding the molecular mechanisms of forma-
tion of different molecular BC subtypes, as well as for
study of possibilities of using CSC as individual mark-
ers of tumors’ malignancy and disease clinical course,
as well as for clarified personalized treatment.
CONCLUSION
Within the luminal A and B, triple negative and HER2-
positive molecular phenotypes of BC the differences
in frequency of cells with CSC markers CD44+/CD24−
were determined. The highest amount of such cells
was observed in BC patients with triple negative (basal)
subtype that can be the cause of its aggressive clinical
course. Correlation between expression of such cells
and presence of metastases in regional lymph nodes
was determined. Difference in overall survival of patients
with BC of basal and molecular subtype depending
on expression in tumor cells of markers of stem cancer
cells CD44+/CD24− was detected.
Detection of tumor cells with markers CD44+/
CD24− within BC molecular subtypes may be addition-
al criterion for the advanced biological characteristic
of BC, and in BC patients with basal molecular sub-
type — for predictive evaluation of individual potential
of tumors to aggressive clinical course.
REFERENCES
1. Cancer in Ukraine, 2012–2013. Bul Nat Cancer Regi ster
of Ukraine. K, 2014: p. 44 (in Ukrainian).
2. Bondar GV, Sedakov IE, Balashova OI, Kho-
menko AV. Assessment of pathomorphological changes
and survival at selective intra-arterial polychemotherapy
of locally disseminated breast cancer. Morphologia 2011;
5: 13–23 (in Russian).
3. Shchepotin IB, Smolanka II, Sklyar SYu, et al. Breast
cancer. Modern aspects of surgical treatment (by data of National
Cancer Institute). Klin Onkol 2013; (1 (9)): 38–43 (in Ukrainian).
4. Smolanka II, Sklyar SYu. Ways of increase of effec-
tiveness of complex treatment of patients with breast cancer.
Kyiv: Drukar, 2007. 207 p. (in Ukrainian).
5. Smolanka II, Sklyar SYu, Ivankova OM, et al. Effec-
tiveness of neoadjuvant polychemotherapy in breast cancer
patients. Treatment pathomorphism. Klin Onkol 2013;
(2 (10)): 1–6 (in Ukrainian).
6. Higgins MJ, Baselga J. Targeted therapies for breast
cancer. J Clin Invest 2011; 121: 3797–803.
7. Marusyk A, Polyak K. Tumor heterogeneity: causes and
consequences. Biochim Biophys Acta 2010; 1805: 105–17.
8. Bertos NR, Park M. Breast cancer — one term, many
entities? J Clin Invest 2011; 121: 3789–96.
9. Sorlie T, Perou CM, Tibshirani R, et al. Gene expression
patterns of breast carcinomas distinguish tumor subclasses with
clinical implications. Proc Natl Acad Sci U S A 2001; 98: 10869–74.
10. Perou CM. Molecular stratification of triple-negative
breast cancers. Oncologist 2010; 15: 39–48.
11. Prat A, Parker JS, Karginova O, et al. Phenotypic and
molecular characterization of the claudin-low intrinsic subtype
of breast cancer. Breast Cancer Res 2010; 12: R68.
Experimental Oncology 37, 58–63, 2015 (March) 63
12. Bertucci F, Finetti P, Cervera N, et al. How different
are luminal A and basal breast cancers? Int J Cancer 2009;
124: 1338–48.
13. Hernandez L, Wilkerson PM, Lambros MB, et al.
Genomic and mutational profiling of ductal carcinomas in situ
and matched adjacent invasive breast cancers reveals intra-
tumour genetic heterogeneity and clonal selection. J Pathol
2012; 227: 42–52.
14. Gerlinger M, Rowan AJ, Horswell S, et al. Intratumor
heterogeneity and branched evolution revealed by multiregion
sequencing. N Engl J Med 2012; 366: 883–92.
15. Martelotto LG, Ng CKY, Piscuoglio S, et al. Breast
cancer intra-tumor heterogeneity. Breast Cancer Res 2014;
16: 210 doi:10.1186/bcr3658.
16. Polyak К. Heterogeneity in breast cancer. J Clin Invest
2011; 121: 3786–8.
17. Zavyalova МV, Denisov EV, Tashireva LA, et al.
Phenotypic drift as a cause for intratumoral morphological
hete rogeneity of invasive ductal breast carcinoma not otherwise
specified. Bio Res Open Access 2013; 2: 148–54.
18. Gottlieb B, Alvarado C, Wang C, et al. Making sense
of intratumor genetic heterogeneity: altered frequency of an-
drogen receptor CAG repeat length variants in breast cancer
tissues. Human Mutation 2013; 34: 610–8.
19. Al-Hajj M, Becker MW, Wicha M. Therapeutic implica-
tion of cancer stem cells. Curr Opin Genet Dev 2004; 14: 43–7.
20. Jordan CT, Guzman ML, Noble M. Cancer stem cells.
N Engl J Med 2006; 355: 1253–61.
21. Guo W, Lasky JL, Wu H. Cancer stem cells. Pediatric
Res 2006; 59: 59R–64R.
22. Pece S, Tosoni D, Confalonieri S, et al. Biological and
molecular heterogeneity of breast cancers correlates with their
cancer stem cell content. Cell 2009; 140: 62–73.
23. Korkaya H, Liu S, Wicha MS. Breast cancer stem cells,
cytokine networks, and the tumor microenvironment. J Clin
Invest 2011; 121: 3804–9.
24. Guttilla IK, Phoenix KN, Hong X, et al. Prolonged
mammosphere culture of MCF-7 cells induces an EMT and
repression of the estrogen receptor by microRNAs. Breast
Cancer Res Treat 2012; 132: 75–85.
25. Chekhun SV, Lukyanova NY, Shvets YV, et al.
Significance of ferritin expression in formation of malignant
phenotype of human breast cancer cells. Exp Oncol 2014;
36: 179–83.
26. Kaur S, Singh G, Kaur K. Cancer stem cells: An insight
and future perspective. J Cancer Res Ther 2014; 10: 846–52.
27. Nguyen LV, Vanner R, Dirks P, et al. Cancer stem
cells: an evolving concept. Nat Rev Cancer 2012; 12: 133–43.
28. Kreso A, Dick JE. Evolution of the cancer stem cell
model. Cell Stem Cell 2014; 14: 275–91.
29. Kim HJ, Kim MJ, Ahn SH, et al. Different prognostic
significance of CD24 and CD44 expression in breast cancer
according to hormone receptor status. Breast 2011; 20: 78–85.
30. Zheng Z, Shao N, Weng H, et al. Correlation between
epidermal growth factor receptor and tumor stem cell markers
CD44/CD24 and their relationship with prognosis in breast
invasive ductal carcinoma. Med Oncol 2015; 32: 275.
31. Balic M, Lin H, Young L, et al. Most early dissemi-
nated cancer cells detected in bone marrow of breast cancer
patients have a putative breast cancer stem cell phenotype. Clin
Cancer Res 2006; 12: 5615.
32. Li W, Liu F, Lei T, et al. The clinicopathological
significance of CD44+/CD24−/low and CD24+ tumor cells
in invasive micropapillary carcinoma of the breast. Pathol Res
Pract 2010; 206: 828–34.
33. Gudadze M, Kankava K, Mariamidze A, et al. Dis-
tribution of cancer stem cells in ductal invasive carcinoma
of breast (review). Georgian Med News 2013; 222: 44–50.
34. Phillips TM, McBride WH, Pajonk F. The response
of CD24(-/low)/CD44+ breast cancer-initiating cells to radia-
tion. J Natl Cancer Inst 2006; 98: 1777–85.
35. Hashimoto K, Shimizu C, Tsuda H, et al. Immuno-
histochemical detection of breast cancer stem cells in hormone
receptor-positive breast cancer and their role in response to endo-
crine therapy and clinical outcome. Oncology 2012; 82: 168–74.
36. Oliveira-Costa JP, Zanetti JS, Silveira GG. Differential
expression of HIF-1α in CD44+CD24−/low breast ductal
carcinomas. Diagn Pathol 2011; 8: 73.
37. Ryabtseva OD, Antipova SV, Lukyanova NYu, et al.
Individual prognosis of serous ovarian cancer survival patients
based on adhesion and proliferation of tumor cells. Oncology
2014; 16: 28–32.
38. Shipitsin M, Lauren L, Campbell LL, et al. Molecular
definition of breast tumor heterogeneity. Cancer Cell 2007;
11: 259–73.
Copyright © Experimental Oncology, 2015
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| id | nasplib_isofts_kiev_ua-123456789-145447 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1812-9269 |
| language | English |
| last_indexed | 2025-12-07T16:57:48Z |
| publishDate | 2015 |
| publisher | Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| record_format | dspace |
| spelling | Chekhun, S.V. Zadvorny, T.V. Tymovska, Yu.O. Anikusko, M.F. Novak, O.E. Polishchuk, L.Z. 2019-01-21T21:21:18Z 2019-01-21T21:21:18Z 2015 СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes / S.V. Chekhun, T.V. Zadvorny, Yu.O. Tymovska, M.F. Anikusko, O.E. Novak, L.Z. Polishchuk // Experimental Oncology. — 2015. — Т. 37, № 1. — С. 58-63. — Бібліогр.: 38 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/145447 Aim: To determine frequency of tumors with immunohistochemical markers of cancer stem cells (CSC) CD44+/CD24− in patients with breast cancer (BC) of different molecular subtype and to evaluate their prognostic value. Object: Surgical material of 132 patients with BC stage I–II, age from 23 to 75 years, mean age — 50.2 ± 3.1 years was studied. Methods: Clinical, immunohistochemical (expression CD44+/CD24−), morphological, statistical. Results: BC is characterized by heterogeneity of molecular subtypes and expression of markers (CD44+/CD24−). Immunohistochemical study of expression of CSC markers in surgical material has detected their expression in 34 (25.4%) patients with BC of different molecular subtypes. The highest frequency of cells with expression of CSC marker was observed in patients with basal molecular subtype (44.8% patients). Most of BC patients with phenotype CD44+/CD24 had stage I of tumor process (34.3%). Statistical processing of data has showen that Yule colligation coefficient equaled 0.28 (р > 0.05) that argues poor correlation between stage of tumor process and number of tumors with positive expression of CSC markers. Statistical processing of data has showen high correlation between presence of cells with expression of CSC markers and metastases of BC in regional lymph nodes (Yule colligation coefficient equals 0.943; р < 0.5). Difference in overall survival of patients with BC of basal molecular subtype depending on expression of CSC CD44+/CD24− markers was detected. Survival of patients with basal BC was reliably higher at lack in tumors of cells with CSC markers CD44+/CD24− and, correspondingly, lower at presence of such cells (р < 0.05). In patients with BC of luminal (A and B), HER-2-positive subtypes, significant change in survival of patients depending on expression of CSC markers was not determined (р > 0.05). Conclusion: Significance of tumor cells with markers CD44+/CD24− within the limits of molecular subtype of BC may be additional criterion for advanced biological characteristic of BC, and in patients with BC of basal molecular subtype — for predictive evaluation of individual potential of tumor to aggressive clinical course. Key Words: breast cancer, cancer stem cells, СD44+/CD24−, molecular subtype. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Original contributions СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes Article published earlier |
| spellingShingle | СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes Chekhun, S.V. Zadvorny, T.V. Tymovska, Yu.O. Anikusko, M.F. Novak, O.E. Polishchuk, L.Z. Original contributions |
| title | СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes |
| title_full | СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes |
| title_fullStr | СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes |
| title_full_unstemmed | СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes |
| title_short | СD44+/CD24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes |
| title_sort | сd44+/cd24− markers of cancer stem cells in patients with breast cancer of different molecular subtypes |
| topic | Original contributions |
| topic_facet | Original contributions |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/145447 |
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