Comprehensive analysis of intratumoral lymphocytes and FOXP3 expression in tumor cells of endometrial cancer
Aim: To study the tumor microenvironment (CD4⁺, CD8⁺ and FOXP3⁺ lymphocytes) and FOXP3 expression by tumor cells and correlation of studied parameters with clinical and morphological characteristics of endometrial adenocarcinomas. Materials and Methods: Tumor samples from 40 patients (mean age 56.9...
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irk-123456789-1453852019-01-22T01:23:34Z Comprehensive analysis of intratumoral lymphocytes and FOXP3 expression in tumor cells of endometrial cancer Lurchenko, N.P. Glushchenko, N.M. Buchynska, L.G. Original contributions Aim: To study the tumor microenvironment (CD4⁺, CD8⁺ and FOXP3⁺ lymphocytes) and FOXP3 expression by tumor cells and correlation of studied parameters with clinical and morphological characteristics of endometrial adenocarcinomas. Materials and Methods: Tumor samples from 40 patients (mean age 56.9 ± 2.8) with endometrial cancer (EC), who did not receive special treatment before surgery (chemotherapy, radiation therapy and hormontherapy), were investigated. Morphological, immunohistochemical methods as well as methods of mathematical statistics were applied in the study. Results: It has been determined that high quantity of FOXP3⁺ tumor cells and intratumoral CD4⁺ and CD8⁺ Т-lymphocytes along with the low content of FOXP3⁺-lymphocytes is typical for the endometrial adenocarcinomas of high differentiation grade (G1). In poorly differentiated (G3) EC an increase of number of FOXP3⁺-lymphocytes and decrease of CD4⁺ and CD8⁺ lymphocytes in lymphocytic infiltrate have been observed. Moreover, decrease of the content of FOXP3⁺ tumor cells has been determined. In EC patients correlation between the following parameters has been detected: proliferative activity and deep invasion of tumor in myometrium (R = 0.74); depth of invasion correlated with the number of the FOXP3⁺ tumor cells (R = −0.63) and number of CD4⁺ and CD8⁺ lymphocytes (R = 0.68 and R = −0.55 respectively) in lymphocytic infiltrate. Thus, results of this study are the evidence of significance of the lymphocytic components of tumor microenvironment and content of FOXP3 expressing tumor cells in EC progression. Conclusion: Quantitative changes of tumor microenvironment, such as number of CD4⁺, CD8⁺ and FOXP3⁺ lymphocytes and content of FOXP3⁺ tumor cells correlate with biological characteristics of EC. Key Words: endometrial cancer, FOXP3⁺ tumor cells, intratumoral FOXP3⁺ lymphocytes, intratumoral CD4⁺, CD8⁺ Т-lymphocytes, depth of invasion, proliferative potential. 2014 Article Comprehensive analysis of intratumoral lymphocytes and FOXP3 expression in tumor cells of endometrial cancer / N.P. Lurchenko, N.M. Glushchenko, L.G. Buchynska // Experimental Oncology. — 2014. — Т. 36, № 4. — С. 262-266. — Бібліогр.: 40 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/145385 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Original contributions Original contributions |
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Original contributions Original contributions Lurchenko, N.P. Glushchenko, N.M. Buchynska, L.G. Comprehensive analysis of intratumoral lymphocytes and FOXP3 expression in tumor cells of endometrial cancer Experimental Oncology |
| description |
Aim: To study the tumor microenvironment (CD4⁺, CD8⁺ and FOXP3⁺ lymphocytes) and FOXP3 expression by tumor cells and correlation of studied parameters with clinical and morphological characteristics of endometrial adenocarcinomas. Materials and Methods: Tumor samples from 40 patients (mean age 56.9 ± 2.8) with endometrial cancer (EC), who did not receive special treatment before surgery (chemotherapy, radiation therapy and hormontherapy), were investigated. Morphological, immunohistochemical methods as well as methods of mathematical statistics were applied in the study. Results: It has been determined that high quantity of FOXP3⁺ tumor cells and intratumoral CD4⁺ and CD8⁺ Т-lymphocytes along with the low content of FOXP3⁺-lymphocytes is typical for the endometrial adenocarcinomas of high differentiation grade (G1). In poorly differentiated (G3) EC an increase of number of FOXP3⁺-lymphocytes and decrease of CD4⁺ and CD8⁺ lymphocytes in lymphocytic infiltrate have been observed. Moreover, decrease of the content of FOXP3⁺ tumor cells has been determined. In EC patients correlation between the following parameters has been detected: proliferative activity and deep invasion of tumor in myometrium (R = 0.74); depth of invasion correlated with the number of the FOXP3⁺ tumor cells (R = −0.63) and number of CD4⁺ and CD8⁺ lymphocytes (R = 0.68 and R = −0.55 respectively) in lymphocytic infiltrate. Thus, results of this study are the evidence of significance of the lymphocytic components of tumor microenvironment and content of FOXP3 expressing tumor cells in EC progression. Conclusion: Quantitative changes of tumor microenvironment, such as number of CD4⁺, CD8⁺ and FOXP3⁺ lymphocytes and content of FOXP3⁺ tumor cells correlate with biological characteristics of EC. Key Words: endometrial cancer, FOXP3⁺ tumor cells, intratumoral FOXP3⁺ lymphocytes, intratumoral CD4⁺, CD8⁺ Т-lymphocytes, depth of invasion, proliferative potential. |
| format |
Article |
| author |
Lurchenko, N.P. Glushchenko, N.M. Buchynska, L.G. |
| author_facet |
Lurchenko, N.P. Glushchenko, N.M. Buchynska, L.G. |
| author_sort |
Lurchenko, N.P. |
| title |
Comprehensive analysis of intratumoral lymphocytes and FOXP3 expression in tumor cells of endometrial cancer |
| title_short |
Comprehensive analysis of intratumoral lymphocytes and FOXP3 expression in tumor cells of endometrial cancer |
| title_full |
Comprehensive analysis of intratumoral lymphocytes and FOXP3 expression in tumor cells of endometrial cancer |
| title_fullStr |
Comprehensive analysis of intratumoral lymphocytes and FOXP3 expression in tumor cells of endometrial cancer |
| title_full_unstemmed |
Comprehensive analysis of intratumoral lymphocytes and FOXP3 expression in tumor cells of endometrial cancer |
| title_sort |
comprehensive analysis of intratumoral lymphocytes and foxp3 expression in tumor cells of endometrial cancer |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| publishDate |
2014 |
| topic_facet |
Original contributions |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/145385 |
| citation_txt |
Comprehensive analysis of intratumoral lymphocytes and FOXP3 expression in tumor cells of endometrial cancer / N.P. Lurchenko, N.M. Glushchenko, L.G. Buchynska // Experimental Oncology. — 2014. — Т. 36, № 4. — С. 262-266. — Бібліогр.: 40 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
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| first_indexed |
2025-07-10T21:33:28Z |
| last_indexed |
2025-07-10T21:33:28Z |
| _version_ |
1837297262457257984 |
| fulltext |
262 Experimental Oncology 36, 262–266, 2014 (December)
COMPREHENSIVE ANALYSIS OF INTRATUMORAL LYMPHOCYTES
AND FOXP3 EXPRESSION IN TUMOR CELLS OF ENDOMETRIAL CANCER
N.P. Iurchenko*, N.M. Glushchenko, L.G. Buchynska
R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology,
National Academy of Siences of Ukraine, Kyiv 03022, Ukraine
Aim: To study the tumor microenvironment (CD4+, CD8+ and FOXP3+ lymphocytes) and FOXP3 expression by tumor cells and
correlation of studied parameters with clinical and morphological characteristics of endometrial adenocarcinomas. Materials
and Methods: Tumor samples from 40 patients (mean age 56.9 ± 2.8) with endometrial cancer (EC), who did not receive special
treatment before surgery (chemotherapy, radiation therapy and hormontherapy), were investigated. Morphological, immunohis-
tochemical methods as well as methods of mathematical statistics were applied in the study. Results: It has been determined that
high quantity of FOXP3+ tumor cells and intratumoral CD4+ and CD8+ Т-lymphocytes along with the low content of FOXP3+-
lymphocytes is typical for the endometrial adenocarcinomas of high differentiation grade (G1). In poorly differentiated (G3)
EC an increase of number of FOXP3+-lymphocytes and decrease of CD4+ and CD8+ lymphocytes in lymphocytic infiltrate have
been observed. Moreover, decrease of the content of FOXP3+ tumor cells has been determined. In EC patients correlation between
the following parameters has been detected: proliferative activity and deep invasion of tumor in myometrium (R = 0.74); depth
of invasion correlated with the number of the FOXP3+ tumor cells (R = −0.63) and number of CD4+ and CD8+ lymphocytes
(R = 0.68 and R = −0.55 respectively) in lymphocytic infiltrate. Thus, results of this study are the evidence of significance
of the lymphocytic components of tumor microenvironment and content of FOXP3 expressing tumor cells in EC progression.
Conclusion: Quantitative changes of tumor microenvironment, such as number of CD4+, CD8+ and FOXP3+ lymphocytes and
content of FOXP3+ tumor cells correlate with biological characteristics of EC.
Key Words: endometrial cancer, FOXP3+ tumor cells, intratumoral FOXP3+ lymphocytes, intratumoral CD4+, CD8+ Т-lymphocytes,
depth of invasion, proliferative potential.
According with the “immune-editing” or “three E”
theory (elimination, equilibrium, escape), immunocom-
petent cells being important component of the tumor
microenvironment can both show antitumor activity
and contribute to the progression of malignant growth
that occurs through selection and survival of the most
aggressive clones of tumor cells resistant to the impact
of cytotoxic T-lymphocytes as well as due to the changes
in functioning of immunocompetent cells in tumor
microenvironment [1–3]. Tumor pathophysiological
features affect the structural and functional changes
of certain components of tumor microenvironment,
in particular decrease or lack the expression of antigen-
recognizing receptors of lymphocytes, antigens of I and
II classes of HLA, increase the quantity of macrophages
of type 2 and T-lymphocytes with regulatory and suppres-
sive activity. These macrophages synthesize appropriate
range of immune suppressing cytokines, which cause
inhibition of effector functions of immunocompetent
cells, in particular CD8+ and CD4+ Т-lymphocytes [4–9].
The main function of regulatory T-lymphocytes
CD4+CD25+FOXP3+ (Tregs) is the maintenance of im-
munologic homeostasis and prevention of autoimmune
diseases. It should be mentioned that in peri pheral
blood of the almost healthy individuals, Tregs consti-
tute only 10% in population of CD4+ Т-lymphocytes.
At the same time, the increase in the quantity of Tregs
up to 30–50% both in peripheral blood and in tumor
microenvironment of patients with cancer of the
diffe rent genesis correlates with unfa vorable clini-
cal course. According to the data of several studies,
particular functional features of Tregs may play key
role in the mechanism of tumor cell escape from
the immune response that contributes to the pro-
gression of malignant process [10–20]. Marker
of Tregs is transcription factor FOXP3+, expression
of which has essential value for the differentiation
of CD4+CD25+FOXP3+ lymphocytes and determines
functions of Tregs regardless of the level of expression
of CD25+ [14–22].
In the most of cases, FOXP3 is mainly expressed
in subpopulation of CD4+CD25+ lymphocytes, but small
quantity of CD8+CD25- cells also express FOXP3 an-
tigen [23].
Until recently, FOXP3 was considered to be expressed
by the population of immunocompetent cells. However,
over recent years, some studies have shown that
FOXP3 is expressed not only in cells of immune system,
but also in tumor cells of different tissues (epithelial and
not epithelial) and is the main factor, which determines
inhibition of proliferative activity of these cells [24, 25].
Besides mentioned above, in tumor cells of many
solid neoplasms, FOXP3+ regulates expression of such
oncogenes and tumor suppressors as C-Myc, ERBB2,
SKP2 and р21WAF1/CIP1 [26, 27].
At the present time, there are only few data about
the role of FOXP3+ lymphocytes in progression of endome-
trial adenocarcinoma [12, 14, 28, 29] and there is no infor-
mation concerning significance of the FOXP3 expression
in the endometrial tumor cells. The question remains
Submitted: July 31, 2014.
*Correspondence: E-mail: laboncogen@yandex.ua
Abbreviations used: EC — endometrial cancer; FOXP3 — Forkhead
box P3.
Exp Oncol 2014
36, 4, 262–266
Experimental Oncology 36, 262–266, 2014 (December) 263
essential concerning the significance of antitumor immu-
nity in the development and clinical course of endometrial
cancer (EC). Also, mechanisms of interaction between
tumor and immunocompetent cells were not clarified.
Relevance of the study is substantiated by the fact
that EC is one of the most widespread malignant
tumors of the female reproductive system. Study
of epidemiological situation in Ukraine has showed that
in female population, EC incidence has significantly
increased over the last 5 years from 15.9 to 17.9%
per 100,000 of the female population and takes third
place (8.6%) after breast cancer and non-melanoma
skin cancer [30].
Considering the mentioned above, the aim
of the study was to investigate the characteristics
of the tumor microenvironment (CD4+, CD8+ and
FOXP3+ lymphocytes) and FOXP3 expression by tumor
cells and correlation of studied parameters with clini-
cal and morphological characteristics of endometrial
adenocarcinoma.
MATERIALS AND METHODS
Samples of surgical material from 40 patients (mean
age 56.9 ± 2.8 years) with EC of FIGO stage I, who did
not receive special treatment before surgery (chemo-
therapy, radiation therapy and hormonal therapy) were
taken for a study. All patients underwent treatment
in oncological gynecology department of the National
Cancer Institute of Ministry of Health of Ukraine. All pa-
tients have given informed consent for the use of their
surgical material for the research purpose.
Assessment of the population of lymphocytes infil-
trating tumor was carried out by immunohistochemistry
using primary monoclonal antibodies: CD4 (clone 4B12,
“Millipore”, USA) and CD8 (clone RIV — 11, “Millipore”,
USA). Expression of FOXP3 was determined both
in tumor cells and in intratumoral lymphocytes using
primary monoclonal antibody to antigen FOXP3 (clone
5H5L12, “Invitrogen”, USA). Proliferative potential of en-
dometrial tumor cells was determined by the number
of cells expressing Ki-67 (clone MIB1, “DakoCytoma-
tion”, Denmark). Results of the immunohistochemical
reaction were assessed by semi-quantitative method
via calculation of the number of positively stained cells
in percentage — labeling index (LI, %). Proliferative
potential was evaluated by determination of the num-
ber of cells expressing protein Ki-67 (proliferation
index — PI). Expression of markers was evaluated
in 800–1000 tumor cells. For corrected interpretation
of the results statistical method of median (Me) determi-
nation was applied: if values of IM and PI were lower than
Me, expression of the corresponding marker was low,
and if values of IM and PI were higher than Me — high.
Statistical analysis was carried out using the software
Statistica 8.0 (StatSoft, Inc.) with use of Kruskal —
Wallis nonparametric criterion. A p value of ≤ 0.05 was
considered to be statistically significant.
RESULTS
Morphological analysis has determined that all studied
tumors were endometrioid adenocarcinomas of different
histological differentiations: 11 (27.5%) tumors were high
(G1), 15 (37.5%) — moderately (G2) and 14 (35.0%) —
poorly differentiated (G3). The distribution of tumors with
respect to the depth of invasion in myometrium was as fol-
lows: 16 (40.0%) tumors invaded less than 1/2 of myome-
trium, 24 (60.0%) neoplasms — more than 1/2.
Immunohistochemical study of the population
of intratumoral T-lymphocytes has revealed that CD4-
lymphocytes were detected in 88.9% and CD8-lympho-
cytes — in 95.0% of studied endometrial tumors (Fig. 1).
a
b
Fig. 1. CD4+ Т-lymphocytes (a) and CD8+ Т-lymphocytes (b)
in the endometrioid adenocarcinoma, × 200
Average number of CD4+ Т-lymphocytes in the lym-
phocytic infiltrate of EC was 35.3 ± 4.1%, and CD8+
Т-lymphocytes — 39.5 ± 4.8%. At the same time,
individual values of quantity of CD4+ and CD8+
Т-lymphocytes in lymphocytic infiltrate of endome-
trial adenocarcinoma significantly varied and consti-
tuted 3.0–48.9% and 5.0–65.0%, respectively. Diffe-
rences in number of CD4+ and CD8+ T-lymphocytes
in the endometrial tumors of different histological
differentiation grade were determined. In G2- and
G3-tumors, the number of intratumoral CD4+ and CD8+
T-lymphocytes decreased as compared to G1 tumors.
It should be mentioned that the number of EC cells
with Ki-67 expression also depended on histologi-
cal differentiation of endometrial adenocarcinoma.
As differentiation grade of EC was decreasing, quan-
tity of endometrial tumor cells expressing Ki-67 was
progressively increasing. In cells of G2 and G3 tumors,
the number of Ki-67 positive tumor cells was higher
264 Experimental Oncology 36, 262–266, 2014 (December)
(p < 0.05) as compared to well differentiated tumors
and constituted 25.6 ± 0.5 and 42.3 ± 0.7%, respec-
tively (Table).
Table. The number of T-lymphocytes in lymphocytic infiltrate and prolife-
rative potential of endometrial adenocarcinomas of different differentia-
tion grade
Tumor dif-
ferentiation
Number of T-lymphocytes, М±m, % The number of cells ex-
pressing Кі-67, М±m, %CD4+ CD8+
G1 52.0 ± 2.7 46.4 ± 5.6 13,4 ± 0.3
G2 35.1 ± 5.1* 40.9 ± 5.6 25.6 ± 0.5*
G3 21.2 ± 5.1* 30.2 ± 4.2 42.3 ± 0.7*
Note: *p < 0.05 as compared to G1.
Expression of FOXP3 has been found in cells
of 75.0% endometrial adenocarcinomas whereas
FOXP3+ intratumoral lymphocytes were detected
in 83.3% tumors (Fig. 2).
a
b
Fig. 2. FOXP3+ lymphocytes (a) and FOXP3 expression by tumor
cells (b) in endometrial adenocarcinoma: a — × 400; b — × 200
The number of cases with positive expression
of FOXP3 by tumor cells decreased from G1 to G3 tu-
mors and reached 100% for G1, 60.0% — for G2 and
50.0% — for G3 endometrial adenocarcinomas whereas
the number of cases with FOXP3+ intratumoral lympho-
cytes in lymphocytic infiltrate conversely increased from
G1 to G3 (G1–62.5%; G2–90.0%; G3–100%) tumors.
At the same time, the number of tumor cells with
FOXP3 expression in G3 was lower (р < 0.03) as com-
pared with G1 adenocarcinomas. In lymphocytic infiltrate
of these tumors, increase (р < 0.04) of the number
of FOXP3+ lymphocytes was observed. These results
show that changes in expression of FOXP3 both in tumor
cells and lymphocytes are associated with morphological
features of endometrial tumors, but oppositely directed
(Fig. 3).
Mean
Mean±SE
Mean±1,96*SE
G1 G2 G3
0
5
10
15
20
25
30
35
40
45
50
55
60
Na
m
be
r o
f l
ym
ph
oc
yt
es
, e
xp
re
ss
in
g
FO
XP
3,
%
Mean
Mean±SE
Mean±1,96*SE
G1 G2 G3
0
5
10
15
20
25
30
35
40
Nu
m
be
r o
f t
um
or
c
el
ls
, e
xp
re
ss
in
g
FO
XP
3,
%
a
b
Fig. 3. Comparison of the number of FOXP3+ lymphocytes (a)
and FOXP3+ tumor cells (b) in endometrial adenocarcinomas
of different differentiation grade: a — Н = 6.487719, р = 0.04;
b — Н = 6.893700, р = 0.03
Detailed analysis of cell population in tumor mi-
croenvironment has determined that at decrease
of differentiation grade in endometrial adenocarci-
nomas, significant decrease of ratio of CD4+/FOXP3+
lymphocytes and CD8+/FOXP3+ lymphocytes was
observed. In G1 tumors, this ratio equaled to 2.3 and
2.1, respectively, in G2 tumors — 1.3 and 1.6, respec-
tively, in G3 tumors — 0.5 and 0.9, respectively. Com-
prehensive assessment of the content of intratumoral
T-lymphocytes, especially quantitative indices of ratio
of subpopulations of CD8+ to FOXP3+ lymphocytes,
gives opportunity to determine their balance that can
be important factor for the determination of the role
of tumor microenvironment in the progression of ma-
lignant growth [31–33].
Thus, at decrease of differentiation grade of en-
dometrial adenocarcinoma, quantitative redistribution
of subpopulation of FOXP3+ lymphocytes towards in-
crease of the number of intratumoral FOXP3+ cells and
decrease of the number of CD4+, CD8+ Т-lymphocytes
occurs. The causes of detected changes can be condi-
tioned, on the one hand, by the impact of such cytokine
as TGF-β1 expressed by most of the tumor cells and tu-
mor-associated macrophages of the II type (M2), which
in turn cause the increased number of Tregs in tumor mi-
croenvironment [34–36]. It is known that Tregs also pos-
sess high secretion of inhibitory cytokines TGF-β, IL-10,
IL-35 and IL-6, which contribute to the decrease or inhi-
bition of functional activity of effector cells, in particular
CD4+ and CD8+ lymphocytes. Intratumoral Tregs produce
Experimental Oncology 36, 262–266, 2014 (December) 265
high level of VEGF that can contribute to the angiogenesis
and growth of tumors. Also, conversion of effector CD4+
and CD8+ Т-lymphocytes in regulatory and suppressive
FOXP3+ Т-lymphocytes is possible [7–10, 13, 22, 37–39].
Regulatory T-lymphocytes can suppress immune reac-
tion not only due to the secretion of cytokines, but also
due to their direct cytotoxic impact [40].
One of the indices associated with progression
of EC is invasive potential. Therefore undoubtedly
important was comparison of the number of cells with
expression of FOXP3+, CD4+ and CD8+ Т-lymphocytes
with proliferative potential and depth of invasion
of endometrial tumor in myometrium. It has been
determined that in the most of well differentiated
endometrial adenocarcinomas (81.8%), IP was lower
than Me and 91.0% of these tumors were characteri-
zed by not deep (< 1/2) invasion in myometrium.
At the same time, in G2 and G3 tumors compared
to G1 tumors, significant increase of the number
of cases with high proliferative potential and deep inva-
sion (> 1/2) in myometrium was observed — (60.0%)
and (54.3%), and (78.6%) and (85.7%), respectively.
It has been determined that in highly prolifera-
ting cells (Кі-67 > 29.0%), number of FOXP3+ tumor
cells (11.4 ± 2.8%; р = 0.01) and CD4+ (25.8 ± 5.9%;
р = 0.008) and CD8+ intratumoral Т-lymphocytes
(31.8 ± 4.3%; р = 0.3) was lower, and number of FOXP3+
intratumoral lymphocytes (41.0 ± 6.7%; р = 0.04) was
higher as compared with tumors with low proliferative ac-
tivity (Кі-67 < 29.0%), in which these indices constituted:
20.8 ± 2.0%; 48.7 ± 4.7%; 41.4 ± 4.3% and 22.0 ± 4.9%,
respectively. Obtained data show significance of expres-
sion of transcription factor FOXP3 in modulation of pro-
liferative potential in endometrial adenocarcinomas. For
instance, increase of number of immunocompetent cells
with expression of FOXP3 contributes to the inhibition
of functions of effector T-lymphocytes, increase of im-
munity suppression and proliferative activity. At the same
time, in tumor cells, impact of FOXP3 protein realized
through the activation of expression of inhibitor of cyclin-
dependent kinases р21WAF1/CIP1 and causes arrest of cel-
lular cycle in S-phase that contributes to the decrease
of proliferative activity of EC cells [26, 27]. It should
be mentioned that the same correlation was detected
in endometrial adenocarcinomas with different invasive
potential of tumor in myometrium (Fig. 4).
For instance, in EC with deep (>1/2) invasion,
significantly lower number of FOXP3+ tumor cells
(10.5 ± 2.0%, р = 0.004), CD4+ lymphocytes (36.5 ±
2.0%, р = 0.03), decreasing tendency of intratumoral
CD8+ lymphocytes (36.0 ± 2.0%, р = 0.3) was ob-
served and number of FOXP3+ lymphocytes (39.1 ±
4.9%, р = 0.01) increased as compared with tumors,
which invaded myometrium less than 1/2 (20.6 ± 2.4;
45.3±5.1; 38.0±4.3 and 15.0±2.9%, correspondingly).
Correlation analysis (Spearman’s rank correla-
tion, р < 0.05) showed strong positive correlation (R
= 0.74) between proliferative activity of cancer cells and
depth of invasion of tumor in myometrium. Moreover,
correlation between depth of invasion of endometrial
adenocarcinoma and number of FOXP3+ tumor cells
(R = −0.63) and components of lymphocytic infiltrate,
such as FOXP3+ and CD4+ lymphocytes, was deter-
mined (R = 0.68 and R = −0.55, correspondingly). De-
termined correlations are the evidence of significance
of components of tumor microenvironment and content
of FOXP3 expressing tumor cells in EC progression.
0
10
20
30
40
50
60
FOXP3+
lymphocytes
CD4+ CD8+ FOXP3+
tumor cells
ІМ
, %
Кі-67 < Ме 29.0%
Кі-67 > Ме 29.0%
0
5
10
15
20
25
30
35
40
45
50
FOXP3+
lymphocytes
CD4+ CD8+ FOXP3+
tumor cells
ІМ
, %
Myometrial Invasion < 1/2
Myometrial Invasion > 1/2
a
b
Fig. 4. Distribution of number of CD4+, CD8+ and FОХP3+ intratu-
moral lymphocytes, and FOXP3+ tumor cells depending on level
of proliferative activity (a) and depth of invasion of EC (b)
Thus, this study determined the changes in the num-
ber of CD4+, CD8+, FOXP3+ lymphocytes and FOXP3+
tumor cells in patients with EC depending on clinical
and morphological features of tumor, as histological
differentiation grade, proliferative potential and depth
of myometrial invasion. It has been demonstrated
that high percentage of intratumoral CD4+ and CD8+
Т-lymphocytes and tumor cells with expression
of FOXP3 as well as decrease of the content of FOXP3+
lymphocytes are associated with high diffe rentiation
grade of endometrioid adenocarcinoma, low expres-
sion of marker of proliferating cells Ki-67 and low
invasion of tumor in myometrium. When differentiation
grade decreases, the increase of population of FOXP3+
lymphocytes in tumor microenvironment is observed
that correlates with decrease of the number of intratu-
moral CD4+ and CD8+ Т-lymphocytes and FOXP3 ex-
pressing tumor cells, high proliferative activity and
deep invasion of tumor in myometrium.
These findings show that quantitative changes
of components of tumor microenvironment, such
as CD4+, CD8+ and FOXP3+ lymphocytes and the con-
tent of FOXP3+ expressing tumor cells objectively
reflect the biological characteristics of EC and may
represent a promising tool toward the identification
of EC patients with poorer prognosis.
266 Experimental Oncology 36, 262–266, 2014 (December)
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