The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium
The hyperplastic processes of the endometrium can arise not only against the background of excessive influence of estrogen, but also against the background of epigenetic damages that affect apoptosis, cell proliferation, differentiation, and adhesion, and DNA reparation. The aim of our study was to...
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| Zitieren: | The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium / V.G. Marichereda, N.A. Bykovа, V.V. Bubnov, G.S. Manasova, T.Y. Moskalenko, A.G. Volyanska, I.M. Shevchenko, T.M. Adamovska // Experimental Oncology. — 2018 — Т. 40, № 2. — С. 109–113. — Бібліогр.: 21 назв. — англ. |
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Marichereda, V.G. Bykovа, N.A. Bubnov, V.V. Manasova, G.S. Moskalenko, T.Y. Volyanska, A.G. Shevchenko, I.M. Adamovska, T.M. 2019-01-24T08:40:07Z 2019-01-24T08:40:07Z 2018 The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium / V.G. Marichereda, N.A. Bykovа, V.V. Bubnov, G.S. Manasova, T.Y. Moskalenko, A.G. Volyanska, I.M. Shevchenko, T.M. Adamovska // Experimental Oncology. — 2018 — Т. 40, № 2. — С. 109–113. — Бібліогр.: 21 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/145574 The hyperplastic processes of the endometrium can arise not only against the background of excessive influence of estrogen, but also against the background of epigenetic damages that affect apoptosis, cell proliferation, differentiation, and adhesion, and DNA reparation. The aim of our study was to investigate and analyze the status of methylation of the promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium. Materials and Methods: The study groups were the following: I — patients with endometrial hyperplasia (EH, n = 9); II — patients with endometrial intraepithelial neoplasia (EIN, n = 10), III — control groups: 1) with endometrial cancer (EC, n = 4), and 2) healthy women (n = 4). Determination of promoter methylation of SFRP2 gene was carried out by the semiquantitative method of methylation-specific PCR assay. Results: The maximum level of methylation of SFRP2 gene promoter had been revealed in patients with EC — 42.80 ± 3.55% (р < 0.05). The patients of the I group had the lowest values of methylation of SFRP2 gene promoter — 10.66 ± 0.85%, while in patients of the II group this indicator was higher — 20.60 ± 0.95% (р < 0.05). In healthy women of the control group, methylation of SFRP2 gene promoter was detected in none of the samples. Conclusion: The content of the methylated SFRP2 gene in endometrial tissue of patients with hyperplastic processes higher than 20–25% allows relate these women to the risk group of EC development and dictates the need of intensive observation of such patients. Key Words: methylation, SFRP2 gene, hyperplastic processes of the endometrium. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Original contributions The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium Article published earlier |
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The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium |
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The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium Marichereda, V.G. Bykovа, N.A. Bubnov, V.V. Manasova, G.S. Moskalenko, T.Y. Volyanska, A.G. Shevchenko, I.M. Adamovska, T.M. Original contributions |
| title_short |
The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium |
| title_full |
The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium |
| title_fullStr |
The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium |
| title_full_unstemmed |
The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium |
| title_sort |
analysis of methylation of dna promoter of sfrp2 gene in patients with hyperplastic processes of the endometrium |
| author |
Marichereda, V.G. Bykovа, N.A. Bubnov, V.V. Manasova, G.S. Moskalenko, T.Y. Volyanska, A.G. Shevchenko, I.M. Adamovska, T.M. |
| author_facet |
Marichereda, V.G. Bykovа, N.A. Bubnov, V.V. Manasova, G.S. Moskalenko, T.Y. Volyanska, A.G. Shevchenko, I.M. Adamovska, T.M. |
| topic |
Original contributions |
| topic_facet |
Original contributions |
| publishDate |
2018 |
| language |
English |
| container_title |
Experimental Oncology |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| format |
Article |
| description |
The hyperplastic processes of the endometrium can arise not only against the background of excessive influence of estrogen, but also against the background of epigenetic damages that affect apoptosis, cell proliferation, differentiation, and adhesion, and DNA reparation. The aim of our study was to investigate and analyze the status of methylation of the promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium. Materials and Methods: The study groups were the following: I — patients with endometrial hyperplasia (EH, n = 9); II — patients with endometrial intraepithelial neoplasia (EIN, n = 10), III — control groups: 1) with endometrial cancer (EC, n = 4), and 2) healthy women (n = 4). Determination of promoter methylation of SFRP2 gene was carried out by the semiquantitative method of methylation-specific PCR assay. Results: The maximum level of methylation of SFRP2 gene promoter had been revealed in patients with EC — 42.80 ± 3.55% (р < 0.05). The patients of the I group had the lowest values of methylation of SFRP2 gene promoter — 10.66 ± 0.85%, while in patients of the II group this indicator was higher — 20.60 ± 0.95% (р < 0.05). In healthy women of the control group, methylation of SFRP2 gene promoter was detected in none of the samples. Conclusion: The content of the methylated SFRP2 gene in endometrial tissue of patients with hyperplastic processes higher than 20–25% allows relate these women to the risk group of EC development and dictates the need of intensive observation of such patients. Key Words: methylation, SFRP2 gene, hyperplastic processes of the endometrium.
|
| issn |
1812-9269 |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/145574 |
| citation_txt |
The analysis of methylation of DNA promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium / V.G. Marichereda, N.A. Bykovа, V.V. Bubnov, G.S. Manasova, T.Y. Moskalenko, A.G. Volyanska, I.M. Shevchenko, T.M. Adamovska // Experimental Oncology. — 2018 — Т. 40, № 2. — С. 109–113. — Бібліогр.: 21 назв. — англ. |
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Experimental Oncology 40, 109–113, 2018 (June) 109
THE ANALYSIS OF METHYLATION OF DNA PROMOTER
OF SFRP2 GENE IN PATIENTS WITH HYPERPLASTIC PROCESSES
OF THE ENDOMETRIUM
V.G. Marichereda1, N.A. Bykovа1, V.V. Bubnov1, G.S. Manasova1, T.Y. Moskalenko2, A.G. Volyanska1,
I.M. Shevchenko1, T.M. Adamovska1
1Odesa National Medical University, Odesa 65082, Ukraine
2Maternity Hospital № 7, Odesa 65082, Ukraine
The hyperplastic processes of the endometrium can arise not only against the background of excessive influence of estrogen, but
also against the background of epigenetic damages that affect apoptosis, cell proliferation, differentiation, and adhesion, and DNA
reparation. The aim of our study was to investigate and analyze the status of methylation of the promoter of SFRP2 gene in patients
with hyperplastic processes of the endometrium. Materials and Methods: The study groups were the following: I — patients with
endometrial hyperplasia (EH, n = 9); II — patients with endometrial intraepithelial neoplasia (EIN, n = 10), III — control groups:
1) with endometrial cancer (EC, n = 4), and 2) healthy women (n = 4). Determination of promoter methylation of SFRP2 gene
was carried out by the semiquantitative method of methylation-specific PCR assay. Results: The maximum level of methylation
of SFRP2 gene promoter had been revealed in patients with EC — 42.80 ± 3.55% (р < 0.05). The patients of the I group had
the lowest values of methylation of SFRP2 gene promoter — 10.66 ± 0.85%, while in patients of the II group this indicator was
higher — 20.60 ± 0.95% (р < 0.05). In healthy women of the control group, methylation of SFRP2 gene promoter was detected
in none of the samples. Conclusion: The content of the methylated SFRP2 gene in endometrial tissue of patients with hyperplastic
processes higher than 20–25% allows relate these women to the risk group of EC development and dictates the need of intensive
observation of such patients.
Key Words: methylation, SFRP2 gene, hyperplastic processes of the endometrium.
Hyperplastic processes of the endometrium (HPE)
are related to the most actual problems in gynecology
and require attention of obstetricians-gynecologists
due to high risk of a malignization and recurrence [1,
2]. HPE is considered to be the pathology of the en-
dometrium, which is characterized by diffuse or focal
enlargement of the endometrium, and can arise not only
at the background of excessive exposure to estrogen,
but also via epigenetic disorders that affect apoptosis,
inactivation of cell proliferation, differentiation, cell ad-
hesion, DNA repair and as a result, lead to an imbalance
of proliferation and apoptosis [3–6].
Also the tendency of “rejuvenation” of this patho-
logy is supervised for last years. It is known, that endo-
metrial cancer (EC) develops against the background
of an endometrial hyperplasia (EH) in about 50%
of cases. Different morphological types of the HPE
(from a simple EH without atypia to an EC) represent
the progressing changes in histo- and cytoarchitecto-
nics with accumulation of the nosospecific molecular
and genetic changes of cells [7, 8].
An abnormal methylation of DNA is the earliest
event, which occurs in cell before the morphological
manifestation of an EH. It is revealed that methylation
of CpG-islands is the cause of loss of an expression
of the corresponding genes due to the blocking of their
transcription [7, 9]. Determination of the status of DNA
methylation of nosospecific genes helps with early and
differential diagnosis of tumors, and also is an impor-
tant event for observation and the prognosis.
The study [10] showed that methylat ion
of FOXP3 gene promoter in patients with EC is around
56–77%. Another study [11] showed that the de-
velopment of HPE is not associated with WIF1 gene
methylation.
SFRPs family secreting proteins are antagonists
of a Wnt-regulatory pathway, they suppress this signal-
ing pathway in healthy individuals. The Wnt-signaling
pathway promotes the regulation of proliferation
and differentiation of cells. It is proved that in some
malignant tumors, such as a colorectal cancer, tu-
mors of head and neck, gastric cancer, methylation
of genes-antagonists of a Wnt-signaling pathway is the
reason of uncontrollable cellular proliferation [12, 13].
Traditionally, ultrasound, endoscopic and patho-
morphological methods of a research are used for
diagnostics of HPE [14]. But none of these methods
shows an exact assessment; even the discrepancy
an atypical hyperplasia and EC in differential diagnos-
tics by morphologists constitutes 11–20% of cases [7].
There is a need for search of more reliable, early cri-
teria of HPE diagnostics by determination of a condi-
tion of DNA methylation. Therefore, research of the
methylation status of SFRP2 gene in patients with HPE
and patients with EC is actual for modern gynecologic
practice.
The aim of our study was to investigate and
analyze the DNA methylation status of the promoter
of SFRP2 gene in patients with HPE.
Submitted: January 16, 2017.
*Correspondence: E-mail: dimitrovanatalie@gmail.com
Abbreviations used: EC — endometrial cancer; EH — endometrial
hyperplasia; EIN — endometrial intraepithelial neoplasia; HPE —
hyperplastic processes of the endometrium; PE — proliferative
endometrium; SE — secretory endometrium.
Exp Oncol 2018
40, 2, 109–113
110 Experimental Oncology 40, 109–113, 2018 (June)
MATERIALS AND METHODS
27 (100%) patients at the age of 18–76 years, have
participated in a research, criteria of inclusion were
the following: the informed patient’s consent to par-
ticipation in a research; age — reproductive, peri- and
postmenopausal; pathomorphologically confirmed
diagnosis of HPE. Criteria of an exclusion were the
age < 18 years or pregnancy.
Involved in the research patients were treated in the
Gynecology Department of Multidisciplinary Medical
Center University Clinic # 1 of Odesa National Medi-
cal University in a planned or urgent order and were
operated thereon due to abnormal uterine bleeding,
HPE. Diagnoses were confirmed by ultrasound dia-
gnostics before an operative intervention. Preoperative
examination and preparation of patients were carried
out by standard principles according to the order
of Ministry of Health Care of Ukraine No. 620 from
29.12.2003. The extent of an operative intervention
was chosen individually for each patient. It depended
on a clinical situation: a hysteroscopy with a biopsy
of endometrium, curettage of a mucous cavity of the
uterus, a hysterectomy with/without uterus appenda-
ges, pipelle biopsy of the endometrium. The existence
of HPE was confirmed pathomorphologically in all pa-
tients who participated in a research according to EIN-
classification of HPE: EH, endometrial intraepithelial
neoplasia (EIN) and classification of histopathologi-
cal subtypes of EC: endometrioid adenocarcinoma,
mucinous carcinoma, serous carcinoma, clear cell
carcinoma, squamous cell carcinoma, undifferentiated
carcinoma, mixed carcinoma, metastatic carcinoma
[1, 15–17].
Depending on the HPE morphological type, exam-
ined patients were distributed into the groups: I) EH —
9 (33.3%) patients; II) EIN — 10 (37.1%) patients;
III) control groups: 1) EC — 4 (14.8%) patients, and
2) healthy women — 4 (14.8%).
The age of patients of the I group was 18–59 years,
among them 6 (66.7%) patients were of reproductive
age, 2 (22.2%) patients — of perimenopausal age,
1 (11.1%) patient of postmenopausal age. 3 (33.3%)
patients, mainly perimenopausal and postmeno-
pausal age, were observed by the therapist because
of an idiopathic hypertension. 2 (22.2%) patients had
high index of body weight — 29.2; 33.4. A hysteroscopy
with a “cold” curettage of endometrium was carried out
to 5 (55.6%) patients, indications to which were sterility
in 2 (40.0%) cases and HPE according to an ultrasound
examination in 3 (60.0%) cases. The hysteroscopy was
carried out with the use of the endoscopic equipment
Karl Storz (Germany). The fractional diagnostic curet-
tage of endometrium was carried out to 3 (33.3%)
women, due to an abnormal uterine bleeding. 1 (11.1%)
patient underwent a hysterectomy without appendages
(basic disease is hysteromyoma with a hemorrhagic
syndrome). USE of organs of the small pelvis, which
was carried out by the device ALOKA Pro Sound SSD
5000 (Japan), showed disharmony of thickness of en-
dometrium to a phase of a menstrual cycle, heteroge-
neity and hyperechogenicity of the endometrium. USE
of organs of a small pelvis had shown the existence
of non-uniform hyperechoic-thickened endometrium
(11–19 mm — in reproductive age, 5–11 mm — in post-
menopausal period). Also it was established that
1 (11.1%) patients had polycystic ovaries, 3 (33.3%) —
hysteromyoma, 1 (11.1%) — adenomyosis.
The age of patients of the II group was 39–64 years:
2 (20.0%) patients of reproductive age and 4 (40.0%)
patients of perimenopausal and postmenopausal age.
5 (50.0%) women had high index of body weight —
28.3–37.4. 5 (50.0%) patients had normal weight. A half
of patients were observed by the therapist because
of a hypertension. 7 (70.0%) patients had appealed
because of menstrual disorders, which turned into ab-
normal uterine bleeding. 3 (30.0%) patients with a dis-
ease of endometrium had no symptoms. USE of organs
of a small pelvis had shown the existence of non-uniform
hyperechoic thickened endometrium (14–23 mm in re-
productive age, 9–17 mm — in postmenopausal period).
Also ultrasound revealed uterine myoma in 5 (50.0%)
women and 1 (10.0%) patient — adenomyosis. 4 (40.0%)
patients had an operative intervention — a hysterectomy
with appendages, 2 (20.0%) women had a hysteroscopic
resection of the endometrium, and 4 (40.0%) patients —
the dilation and curettage of the uterus.
The age of patients of the control group with EC was
49–72 years. 3 (75.0%) patients were at postmeno-
pause, 1 (25.0%) — of reproductive age. The index
of body weight of patients with EC was more than 30, that
confirmed an obesity; 2 (50.0%) patients were on the reg-
ular medical check-up at the therapist because of an id-
iopathic hypertension, 1 (25.0%) — under observation
of the endocrinologist because of a II type diabetes mel-
litus. All patients with EC had complained on menstrual
disorders in the form of a heavy irregular menses and ab-
normal uterine bleedings. 2 (50.0%) patients underwent
the uterus curettage, 2 (50.0%) patients — a hystero-
scopy for biopsy of endometrium. According to results
of USE of organs of the small pelvis, which were carried
out before an operative intervention, all 4 (100%) patients
had the enlargement of the endometrium (in reproduc-
tive age — 18–28 mm, 11–24 mm — at postmenopausal
period), its heterogeneity, in equality of an external con-
tour, absence of accurate margins between the tumor
center and a myometrium. The hysteromyoma had been
diagnosed in 2 (50.0%) patients, and adenomyosis had
been diagnosed in 1 (25.0%) patient.
The healthy women of the control group (4 (100.0%))
without pathology of the endometrium participated
in the study and were examined in the clinic for prepara-
tion to the in vitro fertilization program because of male
infertility factor. The age of the examined control group
was 29–36 years. The absence of the pathology of the
endometrium is confirmed morphologically (pipelle
biopsy of the endometrium) and by ultrasound of the
pelvic organs (M-echo 5–13 mm). Genital and extra-
genital pathology in the control group women were not
detected.
Experimental Oncology 40, 109–113, 2018 (June) 111
Determination of methylation of the DNA promoter
of SFRP2 gene was carried out by the semiquantitative
method of methylation-specific PCR assay.
DNA from samples of endometrium tissues were
isolated with use of DNeasy Blood & Tissue Kit (Qiagen,
USA). Bisulfite processing of DNA was carried out with
using EpiTect Bisulfite Kit (Qiagen, USA) according
to the protocol of the manufacturer. The amount of DNA
for bisulfite processing of DNA in all tests was 1 µg/ml.
Amplification of DNA after bisulfite processing was car-
ried out using Hot Start DNA Polymerase Kit according
to the program 95 °C — 15 min; 95 °C — 30 s, 50 °C — for
a unmethylated DNA and 56 °C — for the methylated
DNA, 72 °C — 30 s, 39 cycles; 72 °C — 10 min (Table 1).
Table 1. The sequence of primers for amplification of the methylation- specific
site of SFRP2 promoter
Type Sequence of primers t, °С
For methylated
SFRP2 gene
F_M_GGGTCGGAGTTTTTCGGAGTTGCGC
R_M_CCGCTCTCTTCGCTAAATACGACTCG
56
For unmethylated
SFRP2 gene
F_U_TTTGGGTTGGAGTTTTTTGGAGTTGTGT
R_U_AACCCACTCTCTTCACTAAATACAACTCA
50
The PCR products were examined using 2%
agarose gel electrophoresis, stained with ethidium
bromide. The relative content the methylated and
unmethylated DNA was assessed using the Quantity
One 1D Analysis Software program (BIO-RAD, USA)
on the device Versa Doc МР 4000 System (BIO-RAD,
USA) according to the formula:
M (%) = A • 100 / (A + B),
where A — is the area of peak of methylated DNA, B —
is the area of peak of unmethylated DNA, M — relative
content the methylated DNA.
The molecular-biological studies of the DNA
methylation were controled by EpiTech Control DNA
unmethylated (Qiagen, USA) and EpiTect Control DNA
methylated (Qiagen, USA).
Statistical analysis of the data obtained was
validated using MedCalc 14.8.1 and Microsoft Excel
2010 packages with the AtteStat add-in 12.5. The
average sample values of the quantitative attributes
are given as M ± m, where M is the mean selective,
m is the mean error. Taking into account the small
number of observations in the groups for comparison
of the main parameters, nonparametric methods were
used — the medication test of Kruskal — Wallis, the
Mann — Whitney test and the Wald — Wolfowitz series.
RESULTS AND DISCUSSION
The analysis of the obtained data on methylation
of the promoter of SFRP2 gene has revealed significant
differences between the patients with EH, EIN, EC and
healthy women (Table 2, Fig. 1, 2).
Table 2. The content of methylated DNA of a gene SFRP2 in the studied
samples of endometrium
Case Morphological type of the
endometrium
The content of SFRP2 gene
methylation, %
1 EH 8
2 EH 12
3 EH 16
4 EH 9
5 EH 11
6 EH 11
7 EH 12
8 EH 8
9 EH 9
10 EIN 22
11 EIN 17
12 EIN 17
13 EIN 19
14 EIN 24
15 EIN 26
16 EIN 23
17 EIN 19
18 EIN 20
19 EIN 19
20 EC 33
21 EC 45
22 EC 49
23 EC 51
24 PE 0
25 PE 0
26 PE 0
27 SE 0
Notе: PE — proliferative endometrium; SE — secretory endometrium.
0
10
20
30
40
50
60
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27
%
Cases
The level of DNA methylation of SFRP2 gene (%)
Fig. 1. The level of methylated SFRP2 gene in the studied endometrial samples. Cases 1–9 — patients with EH, 10–19 — patients
with EIN, 20–23 — patients with EC, 24–27 — healthy women
112 Experimental Oncology 40, 109–113, 2018 (June)
The maximum level of methylation of the stud-
ied gene had been found in patients with EC —
42.80 ± 3.55% (р < 0.05), that is nearly by 4-fold
higher, than in patients with EH and by 2-fold than
in patients with EIN. It was revealed that patients
with EH had the lowest values of this index; the av-
erage content of the methylated SFRP2 gene was
10.66 ± 0.85%, while in patients with EIN this index
was 20.60 ± 0.95% (р < 0.05) (Fig. 1, 2, Table 2).
In healthy women of the control group, methylation
of the investigated gene was detected in none of the
cases, therefore, the graphic representation of the
results was not performed (Table 2).
It is necessary to note that methylation of the pro-
moter of SFRP2 gene didn’t depend on the age of the
patients, existence of the accompanying gynecologic
pathology (hysteromyoma, adenomyosis, polycystic
ovaries).
5
10
15
20
25
30
35
40
45
50
55
Groups
Le
ve
l o
f D
N
A
m
et
hy
la
tio
n
of
th
e
SF
RP
2
ge
ne
, %
EC EH EIN
Fig. 2. Methylation of the promoter of SFRP2 gene in patients
with EH, EIN and EC
Also, an assessment of the intensity of methylation
of the promoter of SFRP2 gene confirmed the results
(Fig. 3) shown above. The methylation of DNA in con-
trol sample (№ 1) was taken as 50%. In comparison
with the control, the minimum intensity of methylation
was noted in patients with EH (samples № 3, № 4,
№ 5). The maximal intensity of DNA methylation was
found in patients with EC (samples № 6, № 7), being
higher in comparison with patients with EH (№ 3, № 4,
№ 5) and the EIN (№ 2).
Fig. 3. Intensity of methylation of the promoter of SFRP2 gene
in endometrial samples from different groups under study. M —
methylated DNA; U — unmethylated DNA
The dependence between the level of DNA methy-
lation and a morphological stage of HPE has shown
its lowest level in patients with EH, and the highest —
in patients with EC. Average values of DNA methylation
were characteristic for patients with EIN.
The data on the status of SFRP2 methylation
in patients with HPE and EC in the literature are
scarce. In the paper of Di Domenico et al. [18] in has
been shown that methylation of SFRP2 gene could
not serve as a diagnostic criterion for the diagnosis
of EC, as it was detected in 2 out of 39 examined
patients with HPE and EC. There is a high incidence
of methylation of another gene from the Secreted
Frizzled Related Protein family, SFRP1, among the
EC specimens and this index allow clearly distinguish
benign and malignant areas of the mucosal tissue
of the uterine cavity. The results of our study showed
a high frequency of SFRP2 gene methylation among
patients with HPE, with a direct relationship between
the degree of methy lation and the morphological type
of the endometrium pathology: EH → EIN → EC. There
are some data that the state of SFRP2 gene methyla-
tion could also be used to diagnosis of oncological
processes of other localizations (colorectal, ovarian,
lung cancer), not only EC [19–21].
CONCLUSION
Thus, the presence of not only morphological
changes in case of HPE, but also characteristic
changes of SFRP2 methylation in different types
of HPE, expands diagnostic criteria of this nosology.
Low content of the methylated DNA (10.66 ± 0.85%)
in case of EH and high content of the methylated
DNA (42.80 ± 3.55%) in patients with EC, allow to use
determination of relative content of the methylated
SFRP2 gene as a marker of differential diagnostics
of HPE and early prognosis of risk of its maligniza-
tion. The SFRP2 gene can be used as a marker for
the differential diagnosis of HPE and EC. The content
of the methylated SFRP2 gene in endometrial tissue
of patients with hyperplastic processes higher than
20–25% allows relate these women to the risk group
of EC development and dictates the need of intensive
observation of such patients.
REFERENCES
1. Tatarchuk TF, Kalugina LV, Tutchenko TМ. Hyperplas-
tic processes of endometrium: what’s new? Repr Endocrinol
2015; 5: 7–13 (in Russian).
2. Sidorova I, Unanyan A, Vlasov R, et al. Endometrial
hyperplastic processes: clinical and therapeutic features. Doc-
tor 2011; 6: 58–60 (in Russian).
3. Gulyaeva LF, Krasilnikov SE. Molecular mechanisms
of endometrial carcinoma. Bull ESSC SD RAMS 2012;
3: 110–5 (in Russian).
4. Delpu Yа, Cordelier P, C. Cho W. DNA methylation and
cancer diagnosis. Int J Mol Sci 2013; 14: 15029–58.
5. Gros C, Fahy J, Halby L, et al. DNA methylation in-
hibitors in cancer: Recent and future approaches. Biochimie
2012; 94: 2280–96.
6. Sivova EN, Pachov АI, Bukreev AV, Fokina AP. Some
questions of etiology and pathogenesis of endometrial prolif-
erative processese. Siber Med Rev 2015; 1: 16–22 (in Russian).
7. Stanoyevich IV, Zemlyakova VV, Fen I, et al. Abnormal
methylation of a number of genes in endometrial hyperplasia
in the presence of chronic endometritis. Russ Bull Obst Gy-
necol 2012; 1: 20–3 (in Russian).
8. Matias-Guiu X, Catasus L, Bussaglia E, et al. Molecular
pathology of endometrial hyperplasia and carcinoma. Hum
Pathol 2001; 32: 569–77.
Experimental Oncology 40, 109–113, 2018 (June) 113
9. Prat J, Gallardo A, Cuatrecasas M, et al. Endometrial
carcinoma: pathology and genetics. Pathology 2007; 39: 72–87.
10. Buchynska LG, Iurchenko NP, Verko NP, et al.
FOXP3 gene promoter methylation in endometrial cancer cells.
Exp Oncol 2015; 4: 246–9.
11. Zaporozhan VN, Dubinina VG, Bubnov VV, et al.
Analysis of gene WIF1 methylation in women with hyperprolif-
erative processes of endometrium and endometrial carcinoma.
Ach Biol Med 2015; 2: 55–7 (in Russian).
12. Ma XY, Ma CX, Wang JN. Endometrial carcinogen-
esis and molecular signaling pathways. Am J Mol Biol 2014;
4: 134–49.
13. Komiya Y, Habas R. Wnt signal transduction pathways.
Organogenesis 2008; 4: 68–75.
14. Zaporozhan VN, Tatarchuk TF, Dubinina VG, et al.
Modern diagnostics and treatment of endometrial hyperplastic
processes. Repr Endocrinol 2012; 1: 5–12 (in Russian).
15. Khanna R, Rupala G, Khanna V, et al. Endometrial
intraepithelial neoplasia and its correlation with WHO classi-
fied endometrial hyperplasia. Int J Pathol 2010; 1: 1–4.
16. Ordi J, Bergeron C, Hardisson D, et al. Reproducibility
of current classifications of endometrial endometrioid glan-
dular proliferations: further evidence supporting a simplified
classification. Histopathology 2013; 64: 284–92.
17. Salman MC, Usubutun A, Boynukalin K, Yuce K. Com-
parison of WHO and endometrial intraepithelial neoplasia clas-
sifications in predicting the presence of coexistent malignancy
in endometrial hyperplasia. J Gynecol Oncol 2010; 21: 97–101.
18. Di Domenico M, Santoro A, Ricciardi C, et al.
Epigenetic fingerprint in endometrial carcinogenesis: the
hypothesis of a uterine field cancerization. Cancer Biol Ther
2011; 5: 447–57.
19. Shunlin L, Xiaoying C, Ruhua C, et al. Diagnostic role
of Wnt pathway gene promoter methylation in non small cell
lung cancer. Oncotarget 2017; 22: 36354–67.
20. Ashktorab H, Brim H. DNA methylation and colorec-
tal cancer. Curr Colorectal Cancer Rep 2014; 10: 425–30.
21. Nikbakht M, Shabanizadeh A, Salehi M, et al.
BRCA1 promoter methylation status in ovarian cancer. Lab
Med 2012; 43: 18–21.
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