Expression OF p53, p21WAF1/CIP1 , p16INK4A and Ki-67 proteins in serous ovarian tumors
Aim: The analysis of p53, p21WAF1/CIP1, p16INK4a and Ki-67 expression in serous ovarian carcinomas of different grade. Materials and Methods: In total, 43 ovarian adenocarcinomas and 8 non-altered ovarian epithelial tissues were immunohistochemically investigated for expression of Кі-67, р53, р21WAF...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2007
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nasplib_isofts_kiev_ua-123456789-1385612025-02-09T20:45:53Z Expression OF p53, p21WAF1/CIP1 , p16INK4A and Ki-67 proteins in serous ovarian tumors Экспрессия белков p53, p21WAF1/CIP1 , p16INK4A И Ki-67 в серозных опухолях яичника Buchynska, L.G. Nesina, I.P. Yurchenko, N.P. Bilyk, O.O. Grinkevych, V.N. Svintitsky, V.S. Original contributions Aim: The analysis of p53, p21WAF1/CIP1, p16INK4a and Ki-67 expression in serous ovarian carcinomas of different grade. Materials and Methods: In total, 43 ovarian adenocarcinomas and 8 non-altered ovarian epithelial tissues were immunohistochemically investigated for expression of Кі-67, р53, р21WAF1/CIP1 and р16INK4a. Results: It has been shown that expression of Кі-67, р53, р21WAF1/CIP1 and р16INK4a in non-altered ovarian epithelial tissue is absent. Serous ovarian carcinomas are characterized by high proliferative activity (PI Кі-67 = 30.0 ± 0.3%), р53 and р16INK4a overexpression (LI is 40.3 ± 0.3% and 31.1 ± 0.6% respectively) and low expression of р21WAF1/CIP1 (LI = 6.8 ± 0.3%). The association between expression of these markers and ovarian tumor grade was defined: the maximal level of Кі-67, р53 and p16/INK4a and minimal of p21WAF1/CIP1 expression were observed in G3 tumors. So, low p21WAF1/CIP1 expression (LI < 7.0%) combined with р16INK4a overexpression is considered to be the factor for a poor prognosis in serous ovarian cancer. Conclusions: The present study has indicated that biomolecular markers of cell proliferation along with traditional clinical and morphologic characteristics can be used for differential diagnostics of ovarian tumors. Цель: анализ экспрессии белков р53, р21WAF1/CIP1, р16INK4a и Кі-67 в серозных опухолях яичника разной степени дифференциации. Материалы и методы: иммуногистохимическое определение уровня экспрессии белков Кі-67, р53, р21WAF1/CIP1 и р16INK4 в образцах операционного материала 43 больных раком яичника и 8 пациенток с фибромиомой матки, эпителиальная ткань яичника которых не изменена. Результаты: установлено, что в неизмененном поверхностном эпителии яичников экспрессия белков Кі-67, р53, р21WAF1/CIP1 и р16INK4a не выявлялась. Для серозных аденокарцином характерна высокая пролиферативная активность (индекс пролиферации (ИП) Кі-67 = 30,0 ± 0,3%), гиперэкспрессия р53 (индекс метки (ИМ) = 40,3 ± 0,3%) и р16INK4 (ИМ = 31,1 ± 0,6%), а также низкий уровень экспрессии р21WAF1/CIP1 (ИМ = 6,8 ± 0,3%). Установлена зависимость уровня экспрессии изученных маркеров от степени дифференцировки серозных опухолей яичника: максимальный уровень экспрессии Кі-67, р53 и p16/INK4a и минимальный p21WAF1/CIP1 отмечали в низкодифференцированных аденокарциномах яичника. Таким образом, низкий уровень экспрессии белка p21WAF1/CIP1 (ИМ < 7,0%) с одновременной гиперэкспрессией р16INK4a можно считать фактором неблагоприятного течения серозного рака яичника. Выводы: результатыпроведенного исследования показали, что молекулярно-биологические маркеры пролиферации клеток наряду с традиционными клиническими и морфологическими характеристиками могут быть использованы для дифференциальной диагностики опухолевого процесса в яичнике. 2007 Article Expression OF p53, p21WAF1/CIP1 , p16INK4A and Ki-67 proteins in serous ovarian tumors / L.G. Buchynska, I.P. Nesina, N.P. Yurchenko, O.O. Bilyk, V.N. Grinkevych, V.S. Svintitsky // Experimental Oncology. — 2007. — Т. 29, № 1. — С. 49-53. — Бібліогр.: 23 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/138561 en Experimental Oncology application/pdf Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| institution |
Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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DSpace DC |
| language |
English |
| topic |
Original contributions Original contributions |
| spellingShingle |
Original contributions Original contributions Buchynska, L.G. Nesina, I.P. Yurchenko, N.P. Bilyk, O.O. Grinkevych, V.N. Svintitsky, V.S. Expression OF p53, p21WAF1/CIP1 , p16INK4A and Ki-67 proteins in serous ovarian tumors Experimental Oncology |
| description |
Aim: The analysis of p53, p21WAF1/CIP1, p16INK4a and Ki-67 expression in serous ovarian carcinomas of different grade. Materials and Methods: In total, 43 ovarian adenocarcinomas and 8 non-altered ovarian epithelial tissues were immunohistochemically investigated for expression of Кі-67, р53, р21WAF1/CIP1 and р16INK4a. Results: It has been shown that expression of Кі-67, р53, р21WAF1/CIP1 and р16INK4a in non-altered ovarian epithelial tissue is absent. Serous ovarian carcinomas are characterized by high proliferative activity (PI Кі-67 = 30.0 ± 0.3%), р53 and р16INK4a overexpression (LI is 40.3 ± 0.3% and 31.1 ± 0.6% respectively) and low expression of р21WAF1/CIP1 (LI = 6.8 ± 0.3%). The association between expression of these markers and ovarian tumor grade was defined: the maximal level of Кі-67, р53 and p16/INK4a and minimal of p21WAF1/CIP1 expression were observed in G3 tumors. So, low p21WAF1/CIP1 expression (LI < 7.0%) combined with р16INK4a overexpression is considered to be the factor for a poor prognosis in serous ovarian cancer. Conclusions: The present study has indicated that biomolecular markers of cell proliferation along with traditional clinical and morphologic characteristics can be used for differential diagnostics of ovarian tumors. |
| format |
Article |
| author |
Buchynska, L.G. Nesina, I.P. Yurchenko, N.P. Bilyk, O.O. Grinkevych, V.N. Svintitsky, V.S. |
| author_facet |
Buchynska, L.G. Nesina, I.P. Yurchenko, N.P. Bilyk, O.O. Grinkevych, V.N. Svintitsky, V.S. |
| author_sort |
Buchynska, L.G. |
| title |
Expression OF p53, p21WAF1/CIP1 , p16INK4A and Ki-67 proteins in serous ovarian tumors |
| title_short |
Expression OF p53, p21WAF1/CIP1 , p16INK4A and Ki-67 proteins in serous ovarian tumors |
| title_full |
Expression OF p53, p21WAF1/CIP1 , p16INK4A and Ki-67 proteins in serous ovarian tumors |
| title_fullStr |
Expression OF p53, p21WAF1/CIP1 , p16INK4A and Ki-67 proteins in serous ovarian tumors |
| title_full_unstemmed |
Expression OF p53, p21WAF1/CIP1 , p16INK4A and Ki-67 proteins in serous ovarian tumors |
| title_sort |
expression of p53, p21waf1/cip1 , p16ink4a and ki-67 proteins in serous ovarian tumors |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| publishDate |
2007 |
| topic_facet |
Original contributions |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/138561 |
| citation_txt |
Expression OF p53, p21WAF1/CIP1 , p16INK4A and Ki-67 proteins in serous ovarian tumors / L.G. Buchynska, I.P. Nesina, N.P. Yurchenko, O.O. Bilyk, V.N. Grinkevych, V.S. Svintitsky // Experimental Oncology. — 2007. — Т. 29, № 1. — С. 49-53. — Бібліогр.: 23 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
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| first_indexed |
2025-11-30T15:29:40Z |
| last_indexed |
2025-11-30T15:29:40Z |
| _version_ |
1850229741628424192 |
| fulltext |
Experimental Oncology ���� ������� ����� ��arc��� ������ ������� ����� ��arc��� ���arc��� ���� �� ��
Nowadays etiology and pat�ogenesis of ovarian
cancer �OC�� remain still unclear. T�e risk factors of t�is
pat�ology include age and peculiarities of reproductive
function�� �ormone factors�� especially estrogens and pro-
gesterone level�� �armful environmental influence [1].
�any of current studies focus on t�e involvement
of oncogenic viruses �t�e Eps�teine-Barr virus�� Hu-
man Papilloma virus type 16�� 18�� �8�� �6�� Herpes virus��
Cytomegalovirus and TT-virus�� and bacterial infec-
tions �Chlamydia trachomatis, Mycoplasma spp.,
Ureaplasma urealiticum�� in t�e occurrence of epit�elial
ovarian tumors [�].
Besides�� referred above suc� genetic factors as
function of tumor suppressor genes and DNA reparation
genes play an important and maybe t�e main role in OC
origin [�]. Application of molecular-biological met�ods
into oncology contributed to forming t�e concept about
presence of certain expression profile in tumors of dif-
ferent origin. Recently�� a group of ovarian tumors wit�
�ig� cell cycle genes expression was distinguis�ed. Ac-
cording to t�e literature data�� about 8�.�% of �ereditary
OC forms are accompanied by alterations in BRCA1 and
BRCA2 genes �in some cases in bot� genes simultane-
ously���� w�ic� are caused by germline and somatic muta-
tions�� loss of �eterozygosity or aberrated met�ylation of
CpG islands in promotor region [�]. �et�ylation of CpG
islands is proved to be one of t�e main ways of inactiva-
tion of suppressor genes in OC [�].
�any aut�ors s�owed t�at mutation of ТР53 tumor
suppressor gene is t�e most frequent genetic feature
in sporadic cancer forms �about ��.�% of cases�� [�].
T�e number of cases wit� mutated ТР53 among ovar-
ian serous carcinomas and endometrioid malignant
tumors is ���.�% and ��.�% respectively wit� maximum
value in poorly differentiated tumors in patients wit� III
or IV stage of disease. Ovarian serous carcinomas �ave
mutated ТР53 gene even at t�e initial stage of illness.
T�e missense mutation in specific DNA-binding domains
leads to cancer development in t�e most cases because
of t�e loss of normal р�� function. W�en wild type and
mutated alleles of ТР53 gene are bot� present in t�e cell��
t�e respective products may form an oligomeric protein
complex. T�is p�enomenon is called “dominant-negative
effect of ТР53 mutation” [6]. In t�is case�� wild type р��
can be only partly inactivated and detected by immuno-
�istoc�emistry. Anot�er type of ТР53 mutation resulting in
t�e loss of gene’s functional activity is premature stop co-
don �nonsense mutation��. Protein products of suc� genes
cannot be detected by immuno�istoc�emistry [��].
One of t�e main р�� effectors is р�1WAF1/CIP1 gene — t�e
cyclin-dependent kinases in�ibitor from CIP/KIP family.
T�e product of t�is gene is р�1WAF1/CIP1 protein�� w�ic� is
able to in�ibit proliferation during almost all cell cycle
p�ases �G1�� S and G���. T�e р�1WAF1/CIP1 protein is acti-
vated by р�� protein encoded by wild�� but not mutant
TP�� type. From ot�er �and�� t�e increase of р�1WAF1/CIP1
expression under t�e presence of mutant ТР�� can
evidence on р��-independent pat�way of regulation of
р�1WAF1/CIP1 expression �e. g. by t�e protein products of
BRCA1, WT1, ТР63 genes or by progesterone�� [8�1�].
Some researc�ers considered t�at t�e c�anges
in p16INK4a gene functions �ave great significance
in OC pat�ogenesis as well [11�1�]. T�e mutations
of t�is gene usually appear on t�e early stages of
transformation of ovarian epit�elium [1�]. First of all��
t�e occurrence of neoplasia is caused by deletion in
�q�1 region�� w�ere р16INK4а gene is located and by
�ypermet�ylation of р16INK4а promotor region. If t�e
mutant gene is �eterozygous�� anot�er allele�� w�ic�
is not mutated�� continues to work as suppressor
and t�e cell can function wit�out abnormalities for a
EXPRESSION OF p53, p21WAF1/CIP1, p16INK4A AND Ki-67 PROTEINS
IN SEROUS OVARIAN TUMORS
L.G. Buchynska1,*, I.P. Nesina1, N.P. Yurchenko1, O.O. Bilyk1, V.N. Grinkevych1, V.S. Svintitsky2
1R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine, Kyiv, Ukraine
2Institute of Oncology, AMS of Ukraine, Kyiv, Ukraine
Aim: The analysis of p53, p21WAF1/CIP1, p16INK4a and Ki-67 expression in serous ovarian carcinomas of different grade. Materials and
Methods: In total, 43 ovarian adenocarcinomas and 8 non-altered ovarian epithelial tissues were immunohistochemically investigated
for expression of Кі-67, р53, р21WAF1/CIP1 and р16INK4a. Results: It has been shown that expression of Кі-67, р53, р21WAF1/CIP1 and
р16INK4a in non-altered ovarian epithelial tissue is absent. Serous ovarian carcinomas are characterized by high proliferative activity
(PI Кі-67 = 30.0 ± 0.3%), р53 and р16INK4a overexpression (LI is 40.3 ± 0.3% and 31.1 ± 0.6% respectively) and low expression
of р21WAF1/CIP1 (LI = 6.8 ± 0.3%). The association between expression of these markers and ovarian tumor grade was defined: the
maximal level of Кі-67, р53 and p16/INK4a and minimal of p21WAF1/CIP1 expression were observed in G3 tumors. So, low p21WAF1/CIP1
expression (LI < 7.0%) combined with р16INK4a overexpression is considered to be the factor for a poor prognosis in serous ovarian
cancer. Conclusions: The present study has indicated that biomolecular markers of cell proliferation along with traditional clinical
and morphologic characteristics can be used for differential diagnostics of ovarian tumors.
Key Words: ovarian adenocarcinoma, tumor grade, Кі-67, р53, p21WAF1/CIP1 and p16/INK4a expression.
Received: March 13, 2007
*Correspondence: Fax: 380(44) 258-16-56
E-mail: lubov@onconet.kiev.ua
Abbreviations used: PI — proliferation index; LI — labeling index;
OC — ovarian cancer.
Exp Oncol �����
���� 1�� �����
�� Experimental Oncology ���� ������� ����� ��arc���
w�ile. However�� �ypermet�ylation of even one allele of
p16INK4а gene makes cell be susceptible to malignant
transformation. Loss of p16INK4a in�ibitory function oc-
curs in ��.�% of serous and 1��.�% of mucinous ovarian
tumors and is more likely for tumor cells wit� wild type
ТР53 (approximately ��.�% of tumors �ave wild type
and �6.�% mutant ТР53�� [11�� 1�].
As ovarian epit�elial cells undergo transformation��
t�ere are not only alteration in ТР53 suppressor gene��
but abnormalities of regulator cell cycle genes as well
�p16INK4a, р21WAF1/CIP1, р27KIP1, р14ARF and ot�ers��.
Taking into consideration all mentioned data�� t�e
aim of our study was to investigate t�e р���� р�1WAF1/CIP1��
р16INK�a and Кі-6�� expression in serous ovarian carci-
nomas of different grade.
MATERIAL AND METHODS
In t�e study�� t�e surgical material of �� patients
wit� OC of I�IV stages was used. T�e age of t�e pa-
tients ranged from �1 to ��6 years�� wit� a mean value
of �1.� ± �.�. T�e unaltered epit�elial ovarian tissue
from 8 women wit� mean age ��.� ± �.� ������� years
old���� w�ic� underwent surgery for uterine fibromyoma��
was used as relative control. All patients were cured in
t�e Department of Oncogynecology�� Institute of On-
cology�� A�S of Ukraine �t�e Head of t�e Department
is prof. L.I. Vorobyova��. T�e informed consent of all
patients �as been received.
Processing of operation material�� immuno�is-
toc�emical detection and evaluation of biomarkers
expression and statistic analysis of obtained results
were accomplis�ed according to t�e standard proce-
dure [1��� 16] wit� modifications of our laboratory [1��].
Histologic diagnosis was performed according to t�e
criteria of t�e World Healt� Organization [18].
T�e operation material was fixed in 1�% neutral
formalin solution and processed for embedding in
paraffin wax. T�e �istologic diagnosis was verified by
studying �ematoxylin & eosin stained sections. T�e
immuno�istoc�emical detection of biomarkers was
conducted using monoclonal primary antibodies �Da-
koCytomation�� Denmark��: anti-Ki-6�� — �IB-1 clone;
anti-р�� — DO-�� clone; anti-р�1WAF1/CIP1 — SX118 clone
and anti-р16INK�a — CINtectm clone. Reaction was visu-
alized by EnVision system wit� next DAB staining. Cell
nuclei were conterstained by �ayer’s �ematoxylin.
T�e results of immuno�istoc�emistry were evaluated
semiquantitatively via calculation of positively stained
cells �labelling index — LI��. T�e expression of markers
was evaluated in 6������� tumor cells. T�e statistic
met�od of median �Ме�� determination for eac� single
parameter was used for correct interpretation of evalu-
ation of t�e biomolecular markers expression. We �ave
found t�at р�1WAF1/CIP1 median is ��.�%�� р16INK�a = 1����%
and р�� = ��.�%. According to t�ese results t�e criteria
for evaluation of markers expression were t�e follow-
ing: t�e protein expression level was considered as low
if LI < 1�.�% for р�� and p16INK�a�� and if LI < ��.�% for
p�1WAF1/CIP1; and as �ig� if 1�.�% ≤ LI < ��.�% for р����
1�.�% ≤ LI < ��.�% for p16INK�a and ��.�% ≤ LI < 1�.�%
for p�1WAF1/CIP1. T�e criteria for protein overexpression
were LI ≥ ��.�% for р��; LI ≥ ��.�% for p16INK�a and
LI ≥ 1�.�% for p�1WAF1/CIP1. T�e proliferative potential
was determined according to t�e number of ʳ-6��-
positive cells: PI < 1�.�% — low proliferative activity��
PI ≥ 1�.�% — �ig� level of proliferation.
RESULTS
By �istologic differentiation�� adenocarcinomas were
subclassified into Grade I �G1�� n = 8���� Grade II �G��� n = 1���
and Grade III �G��� n = ����. Immuno�istoc�emical inves-
tigation s�owed a lack of Кі-6���� р���� р�1WAF1/CIP1 and
р16INK�a expression �except some cases�� in epit�elial
cells of ovarian normal tissue. In contrast to normal
tissue�� t�e expression of t�ese markers was observed
in t�e most of ovarian carcinomas �Table 1��.
Тable 1. The level of expression of biomolecular markers in ovarian
adenocarcinomas with different histologic grade
Pathohis-
tological
diagnosis
The
number
of pa-
tients
The number of positive cells, %,
Ki-67 р53 р21WAF1/CIP1 р16INK4a
Unaltered
ovarian
tissue
8 1.0 (1 case) 0 0 5.0
(1 case)
Adenocar-
cinomas
43 30.0 ± 0.3
(18.0–76.3)
40.3 ± 0.3
(6.7–72.5)
6.8 ± 0.3
(0–31.3)
31.1 ± 0,6
(0–66.7)
G1 8 14.0 ± 0.4
(18.0–28.8)
34.9 ± 0.7
(22.8–38.6)
9.4 ± 0.8
(0–31.3)
11.0 ± 0.8
(0–34.5)
G2 15 32.4 ± 0.5
(18.0–58.7)
37.0 ± 0.5
(12.6–70.0)
8.1 ± 0.8
(0–28.2)
27.0 ± 0.8
(0–42.2
G3 20 37.1 ± 0.4
(20.9–76.3)
45.8 ± 0.5
(6.7–72.5)
3.4 ± 0.3
(0–11.0)
35.9 ± 0.3
(0–66.7)
As s�own in Table 1�� proliferative potential of malig-
nant ovarian tumors as well as р���� р16INK�a expression
were �ig�. �eanw�ile t�e level of p�1WAF1/CIP1 expression
was low �Figure��. Wit� lowering of degree of differen-
tiation ovarian tumor t�e quantity of proliferating cells
increased. Similarly�� t�e increase of t�e quantity of р��
and р16INK�a positive cells was detected in less differenti-
ated ovarian tumors. At t�e same time t�e percentage
of cells wit� p�1WAF1/CIP1 expression in G1 and G� ovarian
adenocarcinomas was low and decreased significantly
in G� tumors �up to ��� �%��.
Expression of some biomolecular markers in OC tu-
mors of certain �istologic grade is s�own in Table �.
Tаble 2. Expression of biomolecular markers in OC tumors
of different grade
The expression
level of biomolecu-
lar markers
Ovarian adenocar-
cinomas (the num-
ber of cases, %)
The grade of differentiation
(the number of cases, %)
G1 G2 G3
р53
Low
High
Overexpression
7.3
19.5
73.2
0
25.0
75.0
6.7
33.3
60.0
11.1
5.6
83.3
р21WAF1/CIP1
Low
High
Overexpression
66.7
25.0
8.3
60.0
20.0
20.0
62.5
25.0
12.5
72.7
27.3
0
р16INK4a
Low
High
Overexpression
25.0
8.3
66.7
75.0
0
25.0
20.0
10.0
70.0
9.1
9.1
81.8
Кі-67
Low
High
11.6
88.4
37.5
62.5
13.3
86.7
0
100.0
T�e most of ovarian adenocarcinomas �88.�%�� were
�ig�-proliferating tumors. T�e p�� expression was ob-
Experimental Oncology ���� ������� ����� ��arc��� �1���� ������� ����� ��arc��� �1�arc��� �1�� �1 �1
served in 1��% of ovarian tumors and was mainly �ig�
and very �ig� �in ��.��% of investigated tumors��.
T�e expression of p�1WAF1/CIP1 protein was revealed
in 6�.�% of tumors�� but t�e �ig� expression level was
detected only in ��.�% and overexpression — in 8.�% of
cases. T�e р16INK�a expression was determined in 8�.�%
of ovarian tumors and its overexpression was prevalent
�66.��% cases��. It is necessary to mark t�e inverse relation
between biomolecular markers overexpression and t�e
grade of ovarian adenocarcinomas differentiation.
T�e decrease of a �istologic grade of ovarian tumor
cells was associated wit� increase in quantity of cases
wit� p�� and р16INK�a overexpression and �ig� level
of proliferative activity�� reac�ing t�e maximum in G�
�8�.��� 81.8 and 1��%�� respectively��. In contrast to it��
t�e number of cases wit� р�1WAF1/CIP1 overexpression
was t�e �ig�est in G1�� decreased in G� and wasn’t
revealed in G� tumors.
DISCUSSION
T�e results of our researc� �ave revealed t�at t�e
malignant ovarian tumors are c�aracterized by t�e
significant proliferative activity and level of р�� and
p16INK�a expression. At t�e same time t�ese tumors were
c�aracterized by low p�1WAF1/CIP1expression. T�e maxi-
mal values of Кі-6���� р�� and p16INK�a and t�e minimal
values of p�1WAF1/CIP1 expression were determinated in
G� ovarian tumors. It is possible t�at suc� decrease
of p�1WAF1/CIP1 expression may be a result of a lack of
transactivating influence of p�� protein on p21WAF1/CIP1
gene. Summing up t�e carried out researc��� it is neces-
sary to mark t�at t�e c�anges of expression of studied
proteins�� w�ic� were determined by us�� coincided wit�
t�e data of ot�er aut�ors [���1�]. In t�ose studies were
described t�at p��-dependent pat�way of p�1WAF1/CIP1
activation is preserved only in clear cell and endometri-
oid ovarian carcinomas but disrupted in t�e most serous
carcinomas�� presumably associated wit� t�e loss of
normal p�� function. In ot�er words�� in serous ovarian
carcinomas t�e activation of p21WAF1/CIP1 gene occurs by
p��-dependent pat�way and expression of p�1WAF1/CIP1
protein determinates t�e activity of Cdk complex — D1-
D�/Cdk��� Cdk6 and proliferative potential of ovarian
cancer�� at t�e same time t�e �ig� proliferative activity
in ovarian tumors caused by t�e low level of p�1WAF1/CIP1
expression [1�]. T�is conclusion is in line wit� our finding
t�at G� tumors wit� a �ig� level of p�� accumulation ex-
press p�1WAF1/CIP1 at low levels. It �as been s�own t�at t�e
patients wit� p�1+р�� - p�enotype �ave more favorable
prognosis t�an t�e patients wit� p�1- р��+ p�enotype��
and t�e determination of p�1WAF1/CIP1/р�� p�enotype is
better prognostic c�aracteristics in patients wit� ovarian
cancer t�an separate determination of level of t�ese
markers expression.
One more reason for derangements of p�1WAF1/CIP1
and р�� proteins functioning can be ВRCA1 gene
Figure. Immuno�istoc�emical detection of expression of ʳ-6�� in G� ���� p�� in G� �b���� p�1WAF1/CIP1 in G� �c�� and p16INK�a in G1 �d��
serous ovarian adenocarcinomas. Original magnification x ��� �a, b, d�� and x ��� �c��
�� Experimental Oncology ���� ������� ����� ��arc���
mutations �trans-activator of ТР53 and p21WAF1/CIP1
genes���� w�ic� are frequent event in �ereditary ovarian
cancer. Alt�oug� t�ey �ave generally been considered
to �ave more limited roles in sporadic ovarian cancer
t�ere are data according to w�ic� t�e mutations at bot�
genes ТР53 and ВRCA1�� w�ic� are associated wit� a
decrease of ВRCA1 expression in sporadic ovarian
cancers are observed [1�]. On t�e ot�er �and�� several
reports �ave focused on cell signal cascades�� w�ic�
occur wit� ТР53 participation and are often interrelated
wit� signal pat�ways t�at are regulated by RB gene.
�utative events or c�anges of pat�ways of ТР53 and
RB are observed in more t�an 8�% of malignant tumors
[��]. It is known t�at some factors can cause t�e RB
inactivation: partial loss of �eterozygosity of RB-locus��
t�e increase of expression of cyclins and cyclin-depen-
dent kinases of serous tumors of ovary is associated
wit� p16INK4a gene overexpression�� probably reflecting
accumulation of inactive p16INK4a products [1�]. It can
be a result of carcinogenic action of Herpes and Hu-
man Papilloma �HPV�� viruses since viral infection con-
tributes to significant increase of p16INK4a expression
[�]. At t�e same time t�ere were data about one more
mec�anism of simultaneous suppression of рRb�� р��
and p�1WAF1/CIP1 proteins by formation of inactivating
complexes between t�em and E6�� E�� HPV oncopro-
teins �for 16 and 18 HPV types�� [��]. T�e functional
in�ibition of Cdk’ in�ibitors by viral oncoproteins may
allow t�ese viruses to promote constitutive activation
of cyclin-Cdk complexes and cell cycle progression.
T�e E6 protein binds and inactivates р�� and E�� — pRb
protein t�at can cause t�e proteolytic degradation of
t�ese suppressors and release of E�F transcriptional
factor. E�� destabilizes pRB by induction its degrada-
tion via an ubiquitin-proteosome pat�way. E�F can
activate p16MTS/INK4a gene transcription and cause sud-
den increase of p16INK�a expression. Besides�� E�� can
also bind and inactivate Cdk in�ibitors p�1WAF1/CIP1 and
p���КIP1�� t�us providing anot�er mec�anism t�roug�
w�ic� E�� can disrupt t�ese cellular processes. E��
also interacts indirectly wit� cyclin E-Cdk��� mediated
t�roug� p1����� pRb-related proteins. T�e significant
distinctions of р�� and p�1WAF1/CIP1 expression were
observed in HPV16-positive and negative tumors in
patients wit� breast cancer [��]. It �as been s�own
t�at t�e level of р�� and p�1WAF1/CIP1expression was sig-
nificantly lower �or was absent�� in t�e most of HPV16-
positive carcinomas and on t�e contrary — HPV16-
negative tumors �ad mainly �ig� expression level of
р�� and p�1WAF1/CIP1 proteins. Considering t�e fact t�at
t�ese viral oncoproteins inactivate and degrade t�e
р�� and p�1WAF1/CIP1 proteins�� t�ese data indicate t�e
resistance of p�� mutant protein to HPV16 action.
In conclusion�� our present data demonstrate t�at
Кі-6���� р���� p�1WAF1/CIP1 and р16INK�a proteins are differ-
ently expressed in normal ovarian surface epit�elium
and ovarian serous adenocarcinomas�� t�at can be
used for differential diagnostics of ovarian malignant
process.
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ЭКСПРЕССИЯ БЕЛКОВ p53, p21WAF1/CIP11/CIP1CIP11, p16INK4A4AA И KKi-67
В СЕРОЗНЫХ ОПУХОЛЯХ ЯИЧНИКА
Цель: анализ экспрессии белков р53, р21WAF1/CIP1, р16INK4a и Кі-67 в серозных опухолях яичника разной степени дифференциации.
Материалы и методы: иммуногистохимическое определение уровня экспрессии белков Кі-67, р53, р21WAF1/CIP11/CIP1CIP11 и р16INK4a4aa в
образцах операционного материала 43 больных раком яичника и 8 пациенток с фибромиомой матки, эпителиальная ткань
яичника которых не изменена. Результаты: установлено, что в неизмененном поверхностном эпителии яичников экспрессия
белков Кі-67, р53, р21WAF1/CIP1 и р16INK4a не выявлялась. �ля серозных аденокарцином характерна высокая пролиферативная ак-�ля серозных аденокарцином характерна высокая пролиферативная ак-
тивность (индекс пролиферации (ИП) Кі-67 = 30,0 ± 0,3%), гиперэкспрессия р53 (индекс метки (ИМ) = 40,3 ± 0,3%) и р16INK4a4aa
(ИМ = 31,1 ± 0,6%), а также низкий уровень экспрессии р21WAF1/CIP11/CIP1CIP11 (ИМ = 6,8 ± 0,3%). �становлена зависимость уровня�становлена зависимость уровня
экспрессии изученных маркеров от степени дифференцировки серозных опухолей яичника: максимальный уровень экспрессии
Кі-67, р53 и p16/INK4a и минимальный p21WAF1/CIP1 отмечали в низкодифференцированных аденокарциномах яичника. Таким
образом, низкий уровень экспрессии белка p21WAF1/CIP1 (ИМ < 7,0%) с одновременной гиперэкспрессией р16INK4a можно считать
фактором неблагоприятного течения серозного рака яичника. Выводы: результаты проведенного исследования показали, что
молекулярно-биологические маркеры пролиферации клеток наряду с традиционными клиническими и морфологическими
характеристиками могут быть использованы для дифференциальной диагностики опухолевого процесса в яичнике.
Ключевые слова: рак яичника, степень дифференцировки, экспрессия биомолекулярных маркеров, Кі-67, р53, p21WAF1/CIP1
и p16/INK4a.
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