Phenotypic features of endometrial tumors in patients with family history of cancer
Aim: To determine the peculiarities of expression of a number of proteins-regulators of the cell cycle in endometrial cancer (EC) cells in patients with a family history of oncological pathologies. Patients and Methods: 95 EC patients (stage І–ІІ) were included into the study. Clinical-genealogical...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
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| Цитувати: | Phenotypic features of endometrial tumors in patients with family history of cancer / L.G. Buchynska, N.P. Iurchenko, N.M. Glushchenko, I.P. Nesina // Experimental Oncology. — 2017 — Т. 39, № 4. — С. 312–318. — Бібліогр.: 41 назв. — англ. |
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irk-123456789-1385482018-06-20T03:04:11Z Phenotypic features of endometrial tumors in patients with family history of cancer Buchynska, L.G. Lurchenko, N.P. Glushchenko, N.M. Nesina, I.P. Original contributions Aim: To determine the peculiarities of expression of a number of proteins-regulators of the cell cycle in endometrial cancer (EC) cells in patients with a family history of oncological pathologies. Patients and Methods: 95 EC patients (stage І–ІІ) were included into the study. Clinical-genealogical analysis was performed. 54 patients (group I) had healthy relatives, and in families of 41 patients (group II) an aggregation of malignant tumors of different genesis (mainly tumors of the gastrointestinal tract and the female reproductive system) was recorded. p53, р21WAF1/CIP1, р16INK4a, and Ki-67 were assessed immunohistochemically in the surgical samples. Results: In the majority of patients, both from group I and II, moderately differentiated tumors were observed (in 38.9 and 46.3% of cases, respectively), mainly with deep myometrium invasion (64.8 and 58.5% of cases, respectively). In EC patients from group II, a significantly higher number of р16INK4a-positive cells (17.7 ± 1.7%; p = 0.001) and lower number of p53-positive (30.9 ± 3.2%; p = 0.05) and Ki-67-positive (26.9 ± 2.7%; p = 0.048) cells was observed compared to those in tumors of patients from group I (12.0 ± 1.6; 37.7 ± 2.8 and 36.7 ± 3.4%, respectively). Conclusion: Phenotypic features of the EC in the patients with family history of cancer differ from those in tumors of patients without such aggregation. The biological heterogeneity of EC seems to relate to the oncogenealogical history of patients. Also this biological heterogeneity is linked to the molecular features of EC cells, which affects cancer aggressiveness and the course of the disease. 2017 Article Phenotypic features of endometrial tumors in patients with family history of cancer / L.G. Buchynska, N.P. Iurchenko, N.M. Glushchenko, I.P. Nesina // Experimental Oncology. — 2017 — Т. 39, № 4. — С. 312–318. — Бібліогр.: 41 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/138548 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 Buchynska, L.G. Lurchenko, N.P. Glushchenko, N.M. Nesina, I.P. Phenotypic features of endometrial tumors in patients with family history of cancer Experimental Oncology |
| description |
Aim: To determine the peculiarities of expression of a number of proteins-regulators of the cell cycle in endometrial cancer (EC) cells in patients with a family history of oncological pathologies. Patients and Methods: 95 EC patients (stage І–ІІ) were included into the study. Clinical-genealogical analysis was performed. 54 patients (group I) had healthy relatives, and in families of 41 patients (group II) an aggregation of malignant tumors of different genesis (mainly tumors of the gastrointestinal tract and the female reproductive system) was recorded. p53, р21WAF1/CIP1, р16INK4a, and Ki-67 were assessed immunohistochemically in the surgical samples. Results: In the majority of patients, both from group I and II, moderately differentiated tumors were observed (in 38.9 and 46.3% of cases, respectively), mainly with deep myometrium invasion (64.8 and 58.5% of cases, respectively). In EC patients from group II, a significantly higher number of р16INK4a-positive cells (17.7 ± 1.7%; p = 0.001) and lower number of p53-positive (30.9 ± 3.2%; p = 0.05) and Ki-67-positive (26.9 ± 2.7%; p = 0.048) cells was observed compared to those in tumors of patients from group I (12.0 ± 1.6; 37.7 ± 2.8 and 36.7 ± 3.4%, respectively). Conclusion: Phenotypic features of the EC in the patients with family history of cancer differ from those in tumors of patients without such aggregation. The biological heterogeneity of EC seems to relate to the oncogenealogical history of patients. Also this biological heterogeneity is linked to the molecular features of EC cells, which affects cancer aggressiveness and the course of the disease. |
| format |
Article |
| author |
Buchynska, L.G. Lurchenko, N.P. Glushchenko, N.M. Nesina, I.P. |
| author_facet |
Buchynska, L.G. Lurchenko, N.P. Glushchenko, N.M. Nesina, I.P. |
| author_sort |
Buchynska, L.G. |
| title |
Phenotypic features of endometrial tumors in patients with family history of cancer |
| title_short |
Phenotypic features of endometrial tumors in patients with family history of cancer |
| title_full |
Phenotypic features of endometrial tumors in patients with family history of cancer |
| title_fullStr |
Phenotypic features of endometrial tumors in patients with family history of cancer |
| title_full_unstemmed |
Phenotypic features of endometrial tumors in patients with family history of cancer |
| title_sort |
phenotypic features of endometrial tumors in patients with family history of cancer |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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2017 |
| topic_facet |
Original contributions |
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http://dspace.nbuv.gov.ua/handle/123456789/138548 |
| citation_txt |
Phenotypic features of endometrial tumors in patients with family history of cancer / L.G. Buchynska, N.P. Iurchenko, N.M. Glushchenko, I.P. Nesina // Experimental Oncology. — 2017 — Т. 39, № 4. — С. 312–318. — Бібліогр.: 41 назв. — англ. |
| series |
Experimental Oncology |
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2025-07-10T06:01:49Z |
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2025-07-10T06:01:49Z |
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1837238652130820096 |
| fulltext |
312 Experimental Oncology 39, 312–318, 2017 (December)
PHENOTYPIC FEATURES OF ENDOMETRIAL TUMORS IN PATIENTS
WITH FAMILY HISTORY OF CANCER
L.G. Buchynska, N.P. Iurchenko*, N.M. Glushchenko, I.P. Nesina
R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine,
Kyiv 03022, Ukraine
Aim: To determine the peculiarities of expression of a number of proteins-regulators of the cell cycle in endometrial cancer (EC) cells
in patients with a family history of oncological pathologies. Patients and Methods: 95 EC patients (stage І–ІІ) were included into
the study. Clinical-genealogical analysis was performed. 54 patients (group I) had healthy relatives, and in families of 41 patients
(group II) an aggregation of malignant tumors of different genesis (mainly tumors of the gastrointestinal tract and the female repro-
ductive system) was recorded. p53, р21WAF1/CIP1, р16INK4a, and Ki-67 were assessed immunohistochemically in the surgical samples.
Results: In the majority of patients, both from group I and II, moderately differentiated tumors were observed (in 38.9 and 46.3%
of cases, respectively), mainly with deep myometrium invasion (64.8 and 58.5% of cases, respectively). In EC patients from group II,
a significantly higher number of р16INK4a-positive cells (17.7 ± 1.7%; p = 0.001) and lower number of p53-positive (30.9 ± 3.2%;
p = 0.05) and Ki-67-positive (26.9 ± 2.7%; p = 0.048) cells was observed compared to those in tumors of patients from group I
(12.0 ± 1.6; 37.7 ± 2.8 and 36.7 ± 3.4%, respectively). Conclusion: Phenotypic features of the EC in the patients with family his-
tory of cancer differ from those in tumors of patients without such aggregation. The biological heterogeneity of EC seems to relate
to the oncogenealogical history of patients. Also this biological heterogeneity is linked to the molecular features of EC cells, which
affects cancer aggressiveness and the course of the disease.
Key Words: endometrial cancer, family history of cancer, biomarkers, p53, р21WAF1/CIP1, р16INK4a, Кі-67.
Endometrial cancer (EC) is one of the most com-
mon malignant neoplasms of the reproductive system
in women, both in Ukraine and in other European coun-
tries. In the structure of oncological pathology of the
female population, according to the National Cancer
Register, in 2015 EC occupied the third place [1]. This
oncopathology occurs predominantly in women over
50 years old, but according to the results of onco-
epidemiological studies in recent years the number
of EC patients in young and middle age has increased,
in particular, 4.0% of cases are diagnosed before the
age of 40, which definitely affects the demographic sit-
uation. It should be noted that at the time of diagnosis
of EC metastases are detected in 17.0% of patients [2].
It has been established that the pathogenesis
of EC, as well as the majority of malignant neoplasms,
is associated with the influence of hereditary factors
and exo- or endogenous factors. On the basis of clini-
cal and genealogical analysis of families of EC patients
from the Kyiv region, the contribution of the genetic
component to the development of EC is 53.2%, in-
dicating the significant importance of hereditary
factors in the occurrence of this pathology [3]. The
incidence of hereditary malignant tumors of the re-
productive system in women, including EC, varies
from 5.0 to 10.0% [4–6]. To date, there are a number
of hereditary cancer syndromes, with a systemic pre-
disposition to the emergence of EC. Most often, EC de-
velops in patients with Lynch syndrome (hereditary
non-polyposis colorectal cancer), the development
of which is associated with inactivation of genes re-
sponsible for the repair of unpaired DNA bases (MSH2,
MLH1, MSH3, MSH6, PMS1, PMS2). In addition, this
oncopatho logy is observed in Lee-Fraumeni syndrome
(mutation in the TP53 gene), Cowden syndrome (a mu-
tation in the PTEN gene), BRCA-associated breast and
ovarian cancer syndrome (BRCA gene inactivation),
and others [4, 7, 8].
Numerous studies have shown that in most cases
EC is hormone-dependent malignancy associated with
the chronic effects of estrogen on the background
of an impaired estrogen-progesterone balance and
is heterogeneous in terms of molecular genetic char-
acteristics that are.
Hyperestrogenemia is one of the factors that modu-
lates the expression of genes, which leads to violations
in the regulation of cellular signals and contributes
to the pathological proliferation of EC cells [9, 10].
The latter is associated with the progression of neo-
plasm: determines the rate of growth, malignancy,
invasive and metastatic potential of neoplasm [11].
Impairment of the proliferation in tumor cells is associ-
ated with a functional imbalance of regulatory genes,
in particular, the oncosuppressor gene ТР53, cyclin-
dependent kinase (CDK) inhibitors р21WAF1/CIP1 and
CDKN2A/р16INK4a [12]. The products of these genes
are the key components of the p53/р21WAF1/CIP1 and
pRb/p16INK4a signaling pathways, inactivation of which
results in uncontrolled proliferation of tumor cells and
determines the biology of malignant tumors [13–18].
Our previous studies have shown that EC is char-
acterized by high proliferative activity, which is due
to changes in expression of the cell cycle regulators
such as p53, р21WAF1/CIP1 and р16INK4a, and is associated
with the tumor differentiation grade [19].
Submitted: October 12, 2017.
*Correspondence: E-mail: laboncogen@gmail.com
Abbreviations used: CDK — cyclin-dependent kinase; EC — endo-
metrial cancer; LI — labeling index; PI — proliferation index.
Exp Oncol 2017
39, 4, 312–318
Experimental Oncology 39, 312–318, 2017 (December) 313
According to current views on the pathogenesis
of tumors of the female reproductive system, genetic
factors play an important role in shaping the biological
characteristics of malignant tumors that may deter-
mine prognosis. According to some authors, sporadic
and hereditary forms of breast and ovarian cancer are
characterized by certain morphological, clinical and
functional differences, in particular, the degree of ma-
lignancy, the hormone receptor phenotype, the course
of the disease, etc. [4, 7, 8]. Despite the achievements
in the field of oncology, to this time in clinical practice
insufficient attention is paid to the assessment of the
role of hereditary factors that determine the biologi-
cal features of malignant neoplasms and the clinical
polymorphism of the disease. Data on the biological
features of endometrial tumors occurring in patients
with a family history of cancer are practically absent
in the modern literature.
Taking into account the above-mentioned, the aim
of the study was to determine the expression of the
cell cycle regulatory proteins in EC of the patients with
a family history of cancer.
MATERIALS AND METHODS
The work was performed on the surgical samples
of 95 EC patients in FIGO stages I or II who did not
receive special treatment before surgery. The patients
were operated in the research department of oncogy-
necology at the National Cancer Institute of the Ministry
of Health of Ukraine. An average age of the patients was
58.3 ± 1.1 years (from 36 to 83 years). Genealogical in-
formation on the burden of a family history of cancer was
obtained in a direct survey of EC patients. All patients
were informed and agreed to use their private informa-
tion and operational material for research.
The study of family history of EC patients was car-
ried out with the help of a specially developed clinical
genealogy card, which included information on the
diseases of relatives of the I and II relation degree, the
patient’s living conditions and related diseases. The
criteria for the distribution of EC patients to the group
with a burdened family history of oncological pathology
were: presence in the proband’s family of two or more
relatives with the I relation degree, or one relative
of the I relation degree and two of the II degree with
neoplasms of the female reproductive system and the
gastrointestinal tract and the earlier age (≤ 50 years)
of the manifestation of the disease [20]. In addition,
information was taken into account on the clinical
course of the disease, the depth of invasion of the tu-
mor in myometrium, the survival rate of patients after
treatment, which was obtained in the study of disease
history and outpatient card.
The morphological diagnosis and tumor differentia-
tion grade were determined on preparations stained
with hematoxylin and eosin according to WHO criteria
(2014) [21].
To determine the cell counts in the phases of the
mitotic cycle, a flow cytometry using flow cytometer
EPICSXL (Becton Coulter, USA). Cell suspension from
surgical EC samples was prepared by mechanical dis-
aggregation of the tissue using a special device called
Medimachine (Becton Dickinson, Italy), followed
by propidium iodide (Sigma-Aldrich, USA) staining.
Immunohistochemical detection of biomarkers was
performed on parallel deparaffined sections using
primary monoclonal antibodies against Ki-67 (clone
MIB-1; Dako, Denmark), p53 (clone DO-7; Dako,
Denmark), р21WAF1/CIP1 (clone HZ52; Dako, Denmark),
р16INK4a (clone JC8; Abcam, UK), and PolyVue HRP/
DAB Detection System (Diagnostic BioSystems, USA).
The results of immunohistochemical reaction were
evaluated by a semi-quantitative method by counting
the percentage of positively stained cells (labeling
index — LI). The proliferative potential was deter-
mined by the number of cells expressing the marker
of proliferation Ki-67 (proliferation index — PI). Ex-
pression of markers was analyzed per 1000 tumor
cells. The significance of the LI and the PI less than
the median was considered to be low for the expres-
sion of the corresponding marker, and with the values
of LI and PI above median — high. It was found that
median expression of Ki-67 in sporadic EC was 31.4%,
in the EC of patients with family history of cancer, the
Ki-67 expression was 29.0%.
The data were processed using the Statistica
7.0 software package (StatSoft, Inc., USA). The follow-
ing statistical methods were used: standard descriptive,
nonparametric (Mann — Whitney, χ2-test), correlation
(Spearman correlation coefficient). p < 0.05 was con-
sidered statistically significant. The analysis of survival
of EC patients was performed using the Kaplan — Meier
method.
RESULTS
The analysis of clinical-genealogical data on pedi-
grees of 54 EC patients (Group I) found no aggregation
of oncological pathology, while in families of 41 pro-
bands (Group II) there was revealed the accumulation
of malignant tumors of different genesis (Table 1).
Most often these were the tumors of the gastrointes-
tinal tract and the female reproductive system.
Table 1. Aggregation of malignant neoplasms in proband families —
patients with EC
Degree
of relation-
ship
Tumor type, n (%)
EC BC OC CGIT LC
Other
localiza-
tions
Total
І
(mother, fa-
ther, sister,
brother, chil-
dren)
10
(15.4)
9
(13.8)
3
(4.6)
20
(30.8)
11
(16.9)
12
(18.5)
65
ІІ
(aunt, uncle,
grandmother,
grandfather)
9
(17.3)
7
(13.5)
– 23
(44.2)
7
(13.5)
6
(11.5)
52
Total 19
(16.2)
16
(13.6)
3
(2.6)
43
(36.8)
18
(15.4)
18
(15.4)
117
Note: *BC — breast cancer; CGIT — cancer of the gastrointestinal tract
(colorectal, stomach, esophagus, liver, pancreas, gallbladder); OC — ovar-
ian cancer; LC — lung cancer.
It should be noted that in families of 24 (58.5%)
EC patients an aggregation of tumors of the colon,
314 Experimental Oncology 39, 312–318, 2017 (December)
stomach, endometrium, and ovary was observed. The
association of EC, breast cancer and ovarian cancer
was found in 10 (24.4%) pedigrees, and in 7 (17.1%)
EC patients was revealed in relatives of the I and II rela-
tion degree [22, 23].
Comparison of the age of manifestation of onco-
logical disease in two groups of patients showed that
in patients with a family history of cancer, EC occurred
at a younger age (55.7 ± 1.5%), compared with those
in patients without aggregation of cancer pathology
in pedigrees (Table 2).
Table 2. Clinical and morphological characteristics of EC patients
depending on a family history of cancer
Clinical and morphological
parameters
Number of EC cases, n (%)
Without aggregation
of oncological patholo-
gy in families,
N = 54
With aggregation
of oncological pa-
thology in families,
N = 41
Average age (range), years 63.2 ± 1.4
(50–76)
55.7 ± 1.5
(26–59)
Tumor differentiation grade:
G1 high
G2 moderate
G3 low
17 (31.5)
21 (38.9)
16 (29.6)
9 (22.0)
19 (46.3)
13 (31.7)
Invasion depth into myome-
trium:
< ½
> ½
19 (35.2)
35 (64.8)
17 (41.5)
24 (58.5)
Morphologically the endometrial carcinomas with
varied differentiation grade and the depth of invasion
were demonstrated in surgical specimens from both
groups of patients (see Table 2). In patients with ag-
gregation of oncopathology in pedigrees, most of the
tumors (46.3%) were moderately differentiated with
an invasion of less than half myometrium (41.5%).
The analysis of the results of the immunohisto-
chemical study of biomarkers in the EC revealed
a significant variability of the individual parameters
both by the number of tumor cells expressing tumor
suppressors and by the proliferative activity (Fig. 1).
In particular, in patients of Group I, the number
of tumor cells with the expression of p53 varied within
the range of 5.3–83.0%, р21WAF1/CIP — 1.6–36.6%,
р16INK4a — 3.0–37.4% and the marker of proliferation
Ki-67 (2.0–89.7%). In EC of patients from Group II,
individual variations in the number of tumor cells with
the expression of these proteins were slightly differ-
ent: p53 (5.1–74.0%), р21WAF1/CIP (2.2–29.0%), р16INK4a
(7.0–39.8%), Ki-67 (9.3–64.2%).
In EC of patients with aggregation of oncologi-
cal pathology in families there was detected a larger
number of cells with expression of protein suppressor
р16INK4a (17.7 ± 1.7%; p = 0.001) and lower numbers
of cells with expression of p53 (30.9 ± 3.2%; p = 0.05)
and the marker of Ki-67 proliferating cells (26.9 ± 2.7%;
p = 0.048), compared to those in tumors of patients
without aggregation of oncological pathology in fami-
lies (12.0 ± 1.6; 37.7 ± 2.8, 36.7 ± 3.4%, respectively)
(Fig. 2). Tumors of Groups I and II of EC patients were
Fig. 1. Immunohistochemical expression of p21WAF1/CIP1 (a), р16INK4a (b), p53 (c), Ki-67 (d) in the endometrial adenocarcinoma.
Counterstained by Mayer hematoxylin. × 400
Experimental Oncology 39, 312–318, 2017 (December) 315
characterized by almost the same number of cells with
expression of CDK inhibitor р21WAF1/CIP.
0
10
20
30
40
50
60
70
р53 p21WAF1/CIP1 p16INK4a Ki-67
Nu
m
be
r o
f p
os
iti
ve
c
el
ls
, %
No family history of cancer
Family history of cancer
Fig. 2. Expression of biomarkers in tumors of EC patients de-
pending on a family history of cancer
It was determined that in the group of patients with
sporadic tumors prevailed high-proliferating (59.0%)
tumors, while in the patients with a family history of can-
cer, the number of such tumors was 43.9% (p < 0.05).
A correlation relationship was established (r = 0.6;
p = 0.01) between the indexes of expression of the
Ki-67 and the p53. That is, the proliferation of sporadic
EC cells significantly increased with high expression
of p53. These data are confirmed by the results of cy-
tofluorometric studies, which showed a significantly
(p < 0.05) higher number of cells (78.9 ± 1.7%) in G0/
G1 and lower in S + G2 + M (21.1 ± 0. 5%) phases of the
cell cycle in carcinomas of patients with an burdened
family history of cancer compared to the patients with
sporadic EC (69.0 ± 1.2 and 31.0 ± 0.7%, respectively).
Thus, EC of patients with a family history of on-
cological pathology is characterized by a lower ex-
pression of the Ki-67 proliferation marker and tumor
suppressor protein p53 with the simultaneous growth
of the expression of the CDK inhibitor р16INK4a.
Analysis of the expression of biomarkers depend-
ing on differentiation grade of the EC showed that the
number of tumor cells with the expression of proteins
Ki-67 and p53 in both groups of patients increased
in the direction from high to moderate and low-dif-
ferentiated tumors. However, the rates of Ki-67 and
p53 were significantly lower in G1 and G3 tumors of pa-
tients with family history of cancer (Table 3).
Table 3. Expression of biomarkers in EC of varying differentiation grade
in patients without aggregation (Group I) and aggregation of tumor patho-
logy in pedigrees (Group II)
Biomarker Groups
of patients
Cell number, % (М ± m)
G1 G2 G3
p53 І 33.2 ± 5.0 31.1 ± 4.5 53.9 ± 5.5*,**
ІІ 18.5 ± 5.4# 27.1 ± 3.8 44.9 ± 5.1*,**
p21WAF1/CIP1 І 7.3 ± 1.1 16.7 ± 4.3*,** 11.4 ± 1.7
ІІ 18.1 ± 2.2# 11.4 ± 1.3 9.2 ± 3.0*
p16INK4a І 17.2 ± 2.8 11.1 ± 1.6 7.2 ± 0.9*
ІІ 12.4 ± 1.3 16.5 ± 1.4# 20.3 ± 3.2*,#
Ki-67 І 24.9 ± 2.4 31.6 ± 4.3 52.9 ± 4.6*,**
ІІ 12.9 ± 2.3# 23.1 ± 2.5 42.1 ± 3.3*,**,#
Note: *p < 0.05 compared with G1 tumors; **p < 0.05 compared with G2 tu-
mors; ***p < 0.05 compared with G1 tumors; #p < 0.05 compared with Group II.
It should be noted that the change in the expres-
sion of the proteins р16INK4a та р21WAF1/CIP1 in EC,
depending on differentiation grades in patients with
burdened and non-burdened family history of cancer,
was divergent. In tumors of patients with EC without
aggregation of oncological pathology in pedigrees,
along with the decrease of differentiation grade,
there was observed a decrease in the number of cells
with expression of protein р16INK4a. At the same time,
in the tumors of patients with a burdened family his-
tory of cancer, the number of cells expressing р16INK4a
protein was significantly (p = 0.03) higher in moder-
ately and low-differentiated carcinomas compared
with EC in Group I patients. The higher number
of cells with expression of р21WAF1/CIP1 was detected
in G2 (16.7 ± 4.3) and G3 tumors compared to G1 tumors
in patients with non-burdened family history of cancer
(p < 0.05).
In carcinomas of patients with EC with a family his-
tory of cancer, a larger number of cells with expression
of р21WAF1/CIP1 protein was observed in highly differentiat-
ed tumors, while with reduced differentiation grade the
number of such cells significantly (p < 0.05) decreased,
reaching the minimum values in low-differentiated
neoplasms compared with those in tumors of patients
without aggregation of oncopathology in family history.
It is known that one of the indicators of progres-
sion of malignant neoplasms is the depth of its inva-
sion in adjacent tissues. Comparison of the expres-
sion of biomarkers in EC with the depth of invasion
in myometrium showed that tumors with a deep (> ½)
invasion of the patients from Groups I and II had a sig-
nificantly higher number of cells with expression of the
p53 protein and Ki-67 proliferation marker compared
to carcinomas without deep (< ½) invasion (Table 4).
Table 4. Expression of biomarkers in EC depending on the depth of tumor
invasion in myometrium and the burden of family history of cancer
Biomarkers
Family history of EC patients
Without aggregation
of oncological pathology
With a family history
of cancer
The depth of invasion of the tumor in myometrium
< ½ > ½ < ½ > ½
Expression of markers, % (М ± m)
p53 36.8 ± 3.1 41.5 ± 4.1 22.8 ± 3.0# 36.9 ± 3.5
p21WAF1/CIP1 9.4 ± 1.7 14.5 ± 2.1 14.3 ± 1.7# 10.4 ± 0.2
p16INK4a 15.9 ± 2.4 9.6 ± 1.4* 14.6 ± 1.4 18.8 ± 0.5**,#
Ki-67 23.5 ± 3.4 48.1 ± 3.7* 18.9 ± 2.4 32.5 ± 2.3**,#
Note: *p < 0.05 compared with tumors with invasion < ½ myometrium in pa-
tients without aggregation of oncological pathology in family; **p < 0.05 com-
pared with in tumors with invasions < ½ of myometrium in patients with a fam-
ily history of cancer; #p < 0.05 compared with tumors in patients without ag-
gregation of oncological pathology in family.
Instead, in tumors that invaded less than ½ of myo-
metrium in patients with EC from Group I, expression
levels of p53 and Ki-67 were higher compared to those
of patients from Group II. It should be noted that the
change in expression of the proteins р16INK4a and
р21WAF1/CIP1 in EC with different depth of invasion of both
groups was similar to the change in the expression
of these proteins, depending on differentiation grade
in these groups. Namely, in tumors with an invasion
> ½ into myometrium, in patients with a non-burdened
family history of cancer, the higher number of cells
with the expression of p21WAF1/CIP1 and lower number
with р16INK4a expression were observed, compared
with those in patients with a family history of cancer.
316 Experimental Oncology 39, 312–318, 2017 (December)
In the study of life expectancy in patients with EC,
depending on the burden of a family history of cancer,
it was found that the overall 5-year survival rate in pa-
tients with EC from families without aggregation of on-
cological pathology was 78.0% and was significantly
lower than in patients with a family history of cancer
(92.0%) (Fig. 3).
0.0
0 10 20 30 40 50
Time (months)
60 70 80 90 100
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
Cu
m
ul
at
ive
p
ro
po
rti
on
s
ur
viv
in
g
No family history of cancer
The 5-year survival rates were 92.0%
The 5-year survival rates were 78.0%
Family history of cancer
Fig. 3. Overall 5-year survival rate of EC patients depending
on a family history of cancer (Kaplan — Meyer, Log-Rank Test,
p = 0.04)
So, the research has shown that in both groups
of patients along with the progression of cancer, i.e.
the decrease of differentiation grade and deep inva-
sion of the tumor in myometrium, the expression of the
Ki-67 and the p53 progressively increased, but the
expression of these markers was significantly more
pronounced in sporadic ECs. Instead, the expression
of CDK inhibitors р16INK4a and p21WAF1/CIP1 was differ-
ently directed, namely, in the tumors of EC patients
with a family history of cancer, there was observed
increased expression of р16INK4a and decreased ex-
pression of p21WAF1/CIP1 in low-differentiated and deeply
invasive tumors compared to moderately and highly
differentiated tumors with invasion ≤ ½ in myometrium.
The phenotypic features of tumors of patients without
aggregation of oncological pathology in pedigrees
and from families with a family history of cancer are
associated with the overall 5-year survival rate of these
patients.
DISCUSSION
Numerous studies have shown that EC course
is characterized by significant variability, which
is determined by the spectrum of genetic disorders,
in particular, suppressor genes whose protein pro-
ducts are involved in various signaling pathways for cell
proliferation control, in particular p53, p21WAF1/CIP1 and
р16INK4a [24, 25].
One of the risk factors for the development of the
tumor process, including EC, is a burdened fam-
ily history of cancer since it is genetic changes that
determine the biology of the tumor modulating the
prognosis of the disease course [4].
In our study, the differences in the clinical course
of the disease in patients without aggregation of on-
cological pathology in pedigrees and family history
of cancer have been established. In particular, signifi-
cantly higher overall 5-year survival (92.0%) was ob-
served in patients with EC with a family history of can-
cer, compared with this in patients without aggregation
of oncological pathology in the pedigree (78.0%).
A similar view is expressed by other researchers, who
noticed a more favorable course of the disease in pa-
tients with hereditary EC, compared to patients with
sporadic EC [23].
Detected differences in the life expectancy
of patients from Groups І and ІІ were associated with
certain molecular-biological characteristics of their
tumors. Thus, in tumors of patients with a burdened
family history of cancer, there was a significantly lower
expression of the p53 protein and the marker of pro-
liferating cells Ki-67 and a higher level of р16INK4a than
in EC of patients without aggregation of oncological
pathology in families. Along with this, tumors of the
studied groups of patients revealed discrepancies
in the expression of CDK inhibitors p21WAF1/CIP1 and
р16INK4a. In patients with a burden family history of can-
cer, there was found a higher number of cells with the
expression of р16INK4a and lower of p21WAF1/CIP1 in low-
differentiated carcinomas and deep invasion of myo-
metrium compared with tumors with high differentia-
tion grade and with invasion < ½. Detected phenotypic
features of tumors of patients with a burdened cancer
history may be due to a specific spectrum of genetic
disorders that occur in various signaling pathways
controlling endometrial cell proliferation, including cell
cycle p53, p21WAF1/CIP1, and р16INK4a.
Possibly, the lower proliferative activity in EC of pa-
tients with a burdened family history of cancer is due
to the effect of suppressor protein p16INK4a, which in-
hibits the CDK4/6 binding to cyclin D1, resulting in pRb
remaining in a hypophosphorylated state in a complex
with E2F and thus blocking G1/S checkpoint. The latter
leads to the accumulation of cells in the G0/G1 phase
of the cell cycle, which we have established during
a cytometric examination of the distribution of en-
dometrial tumor cells by the phases of the mitotic
cycle. The results of the study showed a significant
(p < 0.05) increase in the number of tumor cells in the
G0/G1 phase, in patients with a family history of cancer
compared with sporadic EC.
Taking into account the function of the p16 pro-
tein in the cell cycle, it is possible that its increase
in G3 tumors may be the result of a feedback mecha-
nism in response to the increased proliferative activity
of tumor cells [26].
According to the literature, in the majority of ma-
lignant tumors of different genesis, there are genetic
disorders of the gene TP53 [27]. In EC, the aberrant
expression of p53 is determined by the absence of ex-
pression of its protein product, or its overexpression
(over 50.0% of tumor cells with p53 expression). In this
case, EC with aberrant expression of p53 are charac-
terized by a high degree of malignancy, and the pa-
tients with such tumors showed an unfavorable course
of the disease [28–30]. In our study, it was found that
Experimental Oncology 39, 312–318, 2017 (December) 317
in patients with a burdened family history of cancer, the
number of tumors with the expression of p53 > 50.0%
was significantly lower (χ2 = 3.99; p = 0.04), compared
to the tumors of the patients without aggregation of tu-
mor pathology in pedigree (37.0%).
A small number of tumor cells with p21WAF1/CIP1 pro-
tein expression in low-differentiated tumors of EC pa-
tients with a family history of cancer may result from the
aberrant functioning of the p53 protein that is unable
to activate p21WAF1/CIP1, or activation of p21WAF1/CIP1 is pro-
vided by the p53-independent mechanism.
In contrary, in sporadic tumors, the decrease in the
expression of p16INK4a leads to the activation of a com-
plex of CDK4/6 and cyclin D1, resulting in the phos-
phorylation of Rb and the release of E2F, which acti-
vates the transcription of genes that activate the cel-
lular cycle of S phase, and the Ki-67 increase [31]. The
reason for the decrease in the expression of p16INK4a
in EC cells of patients without aggregation of tumor
pathology in families may be both the methylation
of the promoter of the CDKN2A/p16INK4A gene and
the homozygous deletion of 1 exon of this gene [32].
It has been shown that the expression of the p16INK4a
protein is inversely correlated with differentiation grade
of tumor cells in most endometrioid carcinomas [33],
and in sporadic EC p16INK4A inactivation is more often
observed, especially in cases with a more aggressive
course of the disease [23, 34]. These data are in agree-
ment with the results of our study, which found that
tumors in patients with EC without a burdened family
history of cancer differed by a smaller number of cells
with expression of p16INK4a protein in direct correlation
with survival rates compared to patients with a family
history of cancer.
It could not be excluded that high proliferative activi-
ty in Group I of patients is associated with disturbances
in the signal pathway р53 → p21WAF1/CIP1 ˧ CDK4,6 and
cyclin D, CDK2 and cyclin A, E. The parallel increase
in the number of p53 and Ki-67-positive tumor cells
revealed by us (as confirmed by correlation analysis)
indicates accumulation of mainly functionally aberrant
p53 protein in tumors of patients without an aggre-
gated family history of oncological pathology.
The identified alterations in cell proliferation in the
investigated EC may be associated not only with the
TP53 gene disturbance but also with the inactiva-
tion of one of the regulators of its expression, the
MDM2 protein [35, 36]. According to our previous
studies, with a decrease in EC differentiation grade,
an increase in the expression ratio of p53/MDM2 is ob-
served, which is due to a drastic decrease in the con-
tent of the MDM2 protein. Along with this, the number
of tumors with no MDM2 expression increases [37].
According to the literature, a number of solid tu-
mors possess a high expression of p21WAF1/CIP1, while
retaining the ability to exhibit high proliferative acti-
vity. According to the authors, such tumors may have
specific phosphorylated forms of p21WAF1/CIP1 (T57,
T145, S146 or S130) that have lost their inhibitory
functions [38–40]. That is, the simultaneous growth
of the expression of p21WAF1/CIP1 and Ki-67 proteins
in low-differentiated EC of patients without aggrega-
tion of oncological pathology in pedigrees compared
with their expression in high and moderately differen-
tiated tumors may indicate a lack of inhibitory effect
of p21WAF1/CIP1 protein on proliferation in tumor cells.
Summing up the results of our study we can con-
clude that a more favorable prognosis of patients with
EC with aggregation of tumor pathology in pedigrees
is associated with certain phenotypic peculiarities
of their tumors, which predetermine a lower degree
of malignancy of these tumors, compared with patients
with sporadic EC.
Another mechanism that can explain the more
favorable prognosis of the disease is the presence
of microsatellite instability in the tumors of such
patients, which correlates with a large number of im-
munocompetent cells in the tumor microenvironment,
in particular, intratumoral lymphocytes, which enhance
the immune response [41].
Therefore, the biological heterogeneity of the
EC associated with the hereditary factors is deter-
mined, which modulates molecular-biological features
of the tumor, and influences the EC aggressiveness
and the course of the disease.
REFERENCES
1. Fedorenko ZP, Michailovich YYo, Goulak LO, et al.
Cancer in Ukraine, 2015–2016. Incidence, mortality, activities
of oncological service. Bull Natl Cancer Registry of Ukraine
2017; (18). 86 p.
2. Colombo N, Creutzberg C, Amant F, et al; ESMO-
ESGO-ESTRO Endometrial Consensus Conference Work-
ing Group. ESMO-ESGO-ESTRO Consensus Conference
on Endometrial Cancer: diagnosis, treatment and follow-up.
Ann Oncol 2016; 27: 16–41.
3. Glushchenko NM, Nesina IP, Iurchenko NP, et al. Risk
assessment of cancer of the female reproductive system. Exp
Oncol 2014; 36: 207–11.
4. Wong A, Ngeow J. Hereditary syndromes manifesting
as endometrial carcinoma: how can pathological features aid
risk assessment? Biomed Res Int 2015; 2015: 219012.
5. Imyanitov EN. Screening for persons with hereditary
predisposition to cancer. Pract Oncol 2010; 11: 102–9 (in Rus-
sian).
6. Win AK, Lindor NM, Winship I, et al. Risks of colorec-
tal and other cancers after endometrial cancer for women with
Lynch syndrome. J Natl Cancer Inst 2013; 105: 274–9.
7. Lyubchenko LN, Bateneva YI, Vorotnikov IK, et al.
Hereditary breast cancer: genetic and clinical heterogeneity,
molecular diagnostics, surgical prophylaxis in at-risk groups.
Adv Mol Oncol 2014; 1: 16–25 (in Russian).
8. Lancaster JM, Powell CB, Chen LM, Richardson DL;
SGO Clinical Practice Committee. Society of Gynecologic
Oncology statement on risk assessment for inherited gyneco-
logic cancer predispositions. Gynecol Oncol 2015; 136: 3–7.
9. Tashiro H, Katabuchi H. The relationship between es-
trogen and genes in the molecular pathogenesis of endometrial
cancer. Curr Obstet Gynecol Rep 2014; 3: 9–17.
10. Nishimukai A, Yagi T, Yanai A, et al. High Ki-67 ex-
pression and low progesterone receptor expression could in-
dependently lead to a worse prognosis for postmenopausal
patients with estrogen receptor-positive and HER2-negative
breast cancer. Clin Breast Cancer 2015; 15: 204–11.
318 Experimental Oncology 39, 312–318, 2017 (December)
11. Hanahan D, Weinberg RA. Hallmarks of cancer: the
next generation. Cell 2011; 144: 646–74.
12. Milde-Langosch K, Riethdorf S. Role of cell-cycle
regulatory proteins in gynecological cancer. J Cell Physiol
2003; 196: 224–44.
13. Orlando S, Gallastegui E, Besson A, et al. p27Kip1 and
p21Cip1 collaborate in the regulation of transcription by re-
cruiting cyclin-Cdk complexes on the promoters of target genes.
Nucleic Acids Res 2015; 43: 6860–73.
14. Kitagishi Y, Matsuda S, Minami A, et al. Regulation
in cell cycle via p53 and PTEN tumor suppressors. Cancer
Stud Mol Med Open J 2014; 1: 1–7.
15. Lee EY, Muller WJ. Oncogenes and tumor suppres-
sor genes. Cold Spring Harb Perspect Biol 2010; 2: a003236.
16. Koh VM, Shi YX, Tang QH. P16 and retinoblastoma
protein expression in endometrial carcinoma and clinical sig-
nificance. Eur J Gynaecol Oncol 2011; 32: 309–15.
17. Skirnisdottir I, Seidal T. Association of p21, p21 p27 and
p21 p53 status to histological subtypes and prognosis in low-
stage epithelial ovarian cancer. Cancer Genomics Proteomics
2013; 10: 27–34.
18. Kawamura T, Suzuki J, Wang YV, et al. Linking the
p53 tumour suppressor pathway to somatic cell reprogram-
ming. Nature 2009; 460: 1140–4.
19. Buchynska LG, Nesina IP. Expression of the cell cycle
regulators p53, p21(WAF1/CIP1) and p16(INK4a) in human
endometrial adenocarcinoma. Exp Oncol 2006; 28: 152–5.
20. Committee on Practice Bulletins-Gynecology; Soci-
ety of Gynecologic Oncology. ACOG Practice Bulletin No.
147: Lynch syndrome. Obstet Gynecol 2014; 124: 1042–54.
21. Kurman RJ, Carcangiu ML, Herrington CS,
Young RH. WHO classification of tumours of female repro-
ductive organs, 4th Ed. Lyon: IARC, 2014. 307 p.
22. Lynch PM. Cancer risks after endometrial cancer
in mismatch repair mutation carries. J Natl Cancer Inst 2013;
105: 251–2.
23. Lynch HT, Lynch JF, Attard TA. Diagnosis and man-
agement of hereditary colorectal cancer syndromes: Lynch
syndrome as a model. CMAJ 2009; 181: 273–80.
24. Felix AS, Sherman ME, Hewitt SM, et al. Cell-cycle
protein expression in a population-based study of ovarian and
endometrial cancers. Front Oncol 2015; 5: 25.
25. Le Page C, Huntsman DG, Provencher DM, Mes-
Masson AM. Predictive and prognostic protein biomarkers
in epithelial ovarian cancer: recommendation for future stud-
ies. Cancers (Basel) 2010; 2: 913–54.
26. Saegusa M, Machida BD, Okayasu I. Possible as-
sociations among expression of p14(ARF), p16(INK4a),
p21(WAF1/CIP1), p27(KIP1), and p53 accumulation and
the balance of apoptosis and cell proliferation in ovarian car-
cinomas. Cancer 2001; 92: 1177–89.
27. Kastenhuber ER, Lowe SW. Putting p53 in context.
Cell 2017; 170: 1062–78.
28. Wei JJ, Paintal A, Keh P. Histologic and immunohis-
tochemical analyses of endometrial carcinomas: experiences
from endometrial biopsies in 358 consultation cases. Arch
Pathol Lab Med 2013; 137: 1574–83.
29. Han G, Sidhu D, Duggan MA, et al. Reproducibility
of histological cell type in high-grade endometrial carcinoma.
Mod Pathol 2013; 26: 1594–604.
30. Talhouk A, McConechy MK, Leung S, et al. A clini-
cally applicable molecular-based classification for endometrial
cancers. Br J Cancer 2015; 113: 299–310.
31. Mints M, Mushtaq M, Iurchenko N, et al. Mitochon-
drial ribosomal protein S18-2 is highly expressed in endometri-
al cancers along with free E2F1. Oncotarget 2016; 7: 22150–8.
32. Hu ZY, Tang LD, Zhou Q, et al. Aberrant promoter
hypermethylation of p16 gene in endometrial carcinoma.
Tumour Biol 2015; 36: 1487–91.
33. Koh VM, Shi YX, Tang QH. P16 and retinoblastoma
protein expression in endometrial carcinoma and clinical sig-
nificance. Eur J Gynaecol Oncol 2011; 32: 309–15.
34. Semczuk А, Boltze C, Marzec B, et al. p16INK4A altera-
tions are accompanied by aberrant protein immunostaining
in endometrial carcinomas. J Cancer Res Clin Oncol 2003;
129: 589–96.
35. O’Hara AJ, Bell DW. The genomics and genetics of en-
dometrial cancer. Adv Genomics Genet 2012; 2012: 33–47.
36. Xu H, Zhang Z, Li M, Zhang R. MDM2 promotes
proteasomal degradation of p21Waf1 via a conformation change.
J Biol Chem 2010; 285: 18407–14.
37. Buchynska LG, Nesina IP, Kashuba EV. Different
trends of p53, MDM2 and p14 ARF expression patterns in en-
dometrial adenocarcinomas versus hyperplasia. Exp Oncol
2007; 29: 287–94.
38. Stivala LA, Cazzalini O, Prosperi E. The cyclin-
dependent kinase inhibitor p21CDKN1A as a target of anti-cancer
drugs. Curr Cancer Drug Targets 2012; 12: 85–96.
39. McKenzie PP, Danks MK, Kriwacki RW, Har-
ris LC. P21Waf1/Cip1 dysfunction in neuroblastoma: A novel
mechanism of attenuating G0–G1 cell cycle arrest. Cancer
Res 2003; 63: 3840–4.
40. Child ES, Mann DJ. The intricacies of p21 phosphory-
lation: Protein/protein interactions, subcellular localization
and stability. Cell Cycle 2006; 5: 1313–9.
41. Lynch HT, Drescher KM, de la Chapelle A. Im-
munology and the Lynch syndrome. Gastroenterology 2008;
134: 1246–9.
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