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PROGNOSTIC SIGNIFICANCE OF p53 AND ki67 EXPRESSION IN CERVICAL
CANCER
Khudjamkulova G.I.
xujamkuloveldor916@gmail.com
https://doi.org/10.5281/zenodo.13888971
The aim of this study
is to investigate the prognostic significance of
immunohistochemical expression profiles in patients with cervical cancer who have p53 and
Ki-67 mutations.
Methods:
The paper analyzes data from primary medical records (medical histories and
outpatient records) for 120 women who underwent special treatment at the Republican
specialized scientific and practical medical center of oncology and radiology (RSSPMCOR) of
the Ministry of Health of the Republic of Uzbekistan. A histological examination and analysis
of the content of immunohistochemical markers Ki-67 and p53 in tumor samples (archived
paraffin blocks) was performed.
Results.
A low level of Ki-67 expression was found in 53 (44.2%) patients, intermediate
in 22 (18.3%) and high in 45 (37.5%). Low expression of the mutant protein p53 was detected
in 53 (44.2%) women, moderate in 19 (15.8%) and high in 48 (29%). Proliferative activity
proved to be the most prognostic factor (in terms of the proportion of Ki-67-positive cells), as
with an increase in this indicator by 1 point, the overall survival decreased by 0.99 times. A
multivariate analysis of the combined effect of the studied oncogenes on overall survival,
considering the extent of tumor spread, allowed us to develop a formula for predicting the
risk of disease progression in each individual case.
Conclusion:
The expression of the Ki-67 and p53 markers, along with the extent of
tumor spread, serves as a predictive indicator for cervical cancer prognosis.
Keywords:
cervical cancer, cancer markers, Ki-67, p53, immunohistochemistry.
ПРОГНОСТИЧЕСКАЯ ЗНАЧИМОСТЬ ЭКСПРЕССИИ p53 И ki-67 ПРИ РАКЕ
ШЕЙКИ МАТКИ
Цель.
Изучить прогностическую значимость иммуногистохимического профиля
экспрессии у p53 и ki-67 пациенток, страдающих раком шейки матки.
Методы.
В работе проанализированы данные первичной медицинской
документации (истории болезни и амбулаторные карты) 120 женщинах, получивших
специальное лечение в Республиканском специализированном научно-практическом
медицинском центре онкологии и радиологии (РСНПМЦОиР) Минздрава РУз.
Проведено
гистологическое
исследование
и
изучение
содержания
иммуногистохимических маркёров Ki-67 и р53 в опухоли (архивные парафиновые
блоки).
Результаты.
Низкий уровень экспрессии Ki-67 выявлен у 53 (44,2 %) ,
промежуточный — у 22 (18,3%), высокий у — 45 (37,5%) пациенток. Низкий уровень
экспрессии мутантного протеина p53 диагностирован у 53 (44,2 %), умеренный — у 19
(15,8 %), высокий — у 48 (29%) женщин. Наиболее прогностически значимой оказалась
пролиферативная активность (по доле Ki-67-позитивных клеток), поскольку при
увеличении этого показателя на 1 единицу общая наблюдаемая выживаемость
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снижалась в 0,99 раза, pУалда =0,09. Многофакторный анализ совокупного влияния
изучаемых онкогенов на общую наблюдаемую выживаемость с учётом степени
распространения опухолевого процесса позволил разработать формулу расчёта риска
прогрессирования заболевания в конкретном клиническом наблюдении.
Вывод.
Экспрессия маркёров Ki-67 и р53 совокупности со степенью
распространения опухоли служит прогностическим критерием при раке шейки матки.
Ключевые слова:
рак шейки матки, онкологические маркёры, Ki-67, р53,
иммуногистохимическое исследование.
Introduction
Cervical cancer is one of the most common types of malignant tumors in women,
accounting for a significant proportion of genital cancer cases [3]. Although there have been
advances in the diagnosis and treatment of cervical cancer, the incidence of the disease has
been increasing, as has its aggressiveness [1].
Currently, there is a lot of attention being paid to the study of the effect of the level of
expression of various antigens (Ki-67, p53, cyclin E, p150, metalloproteinases, and other
factors related to proliferative activity, apoptosis, angiogenesis, and so on) on the
effectiveness of cervical cancer treatment.
One of the most widely studied indicators of tumor aggressiveness is cellular
proliferation. This can be assessed by measuring the mitotic index and the level of Ki-67
expression. Ki-67 is a nuclear protein that is expressed throughout the cell cycle except for the
G0 phase and the early stages of G1. The proliferative index, which is the level of Ki-67
expression, can serve as an independent predictor of recurrence, overall survival, relapse-free
survival, and response to chemotherapy and radiation therapy in various tumor types.
The presence of functional and structural abnormalities in cell cycle regulators, such as
cyclins and cyclin-dependent kinases, has been identified in cells from various types of
cancerous tumors. A higher expression level of the cyclin B1 gene was observed in cells from
invasive cervical cancer compared to cells from normal tissue (p = 0.019), while there were no
significant differences in the level of expression of cyclin D1.
According to individual studies, overexpression of cyclin D1 has been found to be an
independent unfavorable prognostic factor in cervical cancer, regardless of its stage and
histological variant.
However, the significance of immunohistochemical marker evaluation in cervical
cancer remains poorly understood, and there is no consensus on the role of Ki-67, Her2/neu,
Bcl-2 and p53 expression as prognostic factors, nor on their combination with other clinical
and morphological tumor features [7, 8, 10]. This study aims to investigate the prognostic
significance of antigen expression profiles in patients with cervical cancer.
Methods and materials.
The data from the primary medical records (medical history and outpatient records) for
120 women who received specialized treatment at the Republican specialized scientific and
practical medical center of oncology and radiology (RSSPMCOR) of the Ministry of Health of
the Republic of Uzbekistan were analyzed. Histological analysis and expression of
immunohistochemistry markers Ki-67 and p53 were performed on tumor tissue. Of the total
number of patients, 39 had Stage I cervical cancer, 31 had Stage II cervical cancer, 26 had
Stage III cervical cancer, and 4 had Stage IV cervical cancer.
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The age group of women in the first stage of maturity included 15 individuals, the
second stage of maturity - 69 individuals, and the elderly age group - 16 individuals .
According to the histological examination, 94 patients had squamous cell carcinoma and 6
patients had adenocarcinoma.
Immunohistochemical analysis of the expression of p-53 and Ki-67 tissue antigens was
performed using commercial monoclonal antibodies from DakoCytomation (Denmark).
Before conducting the immunohistochemical study, a staining method was developed by
selecting the optimal primary antidiv dilution, exposure time, and unmasking buffer.
Serial sections with a thickness of 5 microns were prepared from archival paraffin
blocks. These were then placed on Super Frost precut slides (Menzel-Glaser, Germany) and
allowed to dry at room temperature for the day. Before staining, the sections were placed
upright in a thermostat at 60 °C for 60 minutes.
After this, they were refined in orthoxylene for 10 minutes in a battery of two tanks.
They were then rehydrated in ethyl alcohol in a descending concentration for 3 minutes in a
series of three tanks. Finally, they were rinsed in distilled water. The slides with sections were
then transferred to a heated unmasking buffer (Target Retrieval Solution) and placed in a
water bath at 98 °C for 20-40 minutes. Once cooled to room temperature, they were washed
2-3 times with an aqueous solution with a pH of 7.4. To block endogenous peroxidase activity,
the sections were treated with a 3% hydrogen peroxide solution for 15 minutes.
Results
As a result of the studies conducted, it was found that the level of expression of the Ki-67
tumor proliferation activity antigen varied from 0% to 98%, and the level of mutant protein
p53 varied from 0% to 100%.
A low level of Ki-67 expression was found in 44% of patients, intermediate in 19%, and
high in 37%. At the same time, low proliferative activity was diagnosed in two women with
stage I cervical cancer, 10 with stage II, and 8 with stage III. Moderate levels were noted in 19
cases at stage I, 5 at stage II, and 8 at stage III. High antigen expression was detected in eight
patients with stage I cancer, 16 at stage II, and 10 at stage III (p<0.05). The average Ki-67 level
in stage I was 23.03% ±0.52, stage II - 52.94% ±34.5, and stage III - 44.81% ±34.17 (p<0.05).
There were significant differences between stages I and II (p<0.005) and between stages I and
III (p<0.01), but not between stages II and III (p>0.05).
Low expression of the mutant p53 protein was observed in 44 (55%) women, moderate
in 16 (16%) and high in 29 (29%). Low expression of p53 was seen in 34 patients with stage I
cervical cancer, 14 at stage II and 7 at stage III. High levels of the mutant p53 protein were
seen in 1 patient at each stage (I, II and III), respectively (p < 0.05).
The average value of p53 level was 6.03 ± 13.42% in stage I, 36.58 ± 34.53% in stage II
and 53.08 ± 32.21% in stage III (p < 0.05). Statistical significance of the difference between
the average expression of p53 between stages I and II (p < 0.05), I and III (p < 0.05) was
revealed, but there was no significant difference between stage II and III levels (p > 0.05).
Discussion
Then, by substituting the values of the coefficients, the risk of cervical
cancer progression was calculated for each of the patients who underwent
immunohistochemical testing (120 patients). Based on previously obtained data on
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median survival time in 120 patients with stage I-IV cervical cancer, they were divided into
groups with different prognoses.
The value of the progression probability coefficient in stage II of the disease ranged from
1.596 to 4.084, with an average value of 3.307. In cases where death occurred, this indicator
ranged from 2.946 to 3.878, with an average value of 3.237. Out of the 31 patients diagnosed
with stage II cervical cancer, 10 (32.26%) passed away, and in nine of these cases, the median
life expectancy was below 87 months, with values ranging from 6 to 32 months.
The risk factor for disease progression in stage III had a value
between 2.427 and 4.7, with an average value of 3.814. For death, the indicator ranged from
2,538 to 4,392, with an average of 3,690. Out of the 26 patients with stage III cervical cancer,
13 (50%) died. Of these 13 deaths, 8 had a median life span below 18 months, with a
range of 4 to 13 months.
Statistically significant differences were found between the average values of the Ki-67
nuclear antigen expression indicators in patients with stages I and II of the disease, as well as
between stages I and III (p <0.05). However, no significant differences were observed
between the average marker values at stages II and III (stage I - 23.03±0.52%, stage II -
52.94±34.5%, stage III - 44.81±34.17%).
An increase in p53 expression was observed with an increase in tumor process
prevalence (the average p53 level value at stage I was 6.03±13.42%, at stage II -
36.58±34.53%, and at stage III - 53.08 ±32.21%, p<0.05).
Conclusion.
The expression of Ki-67 and p53 markers, in combination with the extent of
tumor spread, serves as a prognostic indicator for cervical cancer.
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