Dynamics of saliva cytokine levels during intraoperative photodynamic therapy in patients with locally advanced oral cancer Текст научной статьи по специальности «Клиническая медицина»

South Russian Journal of Cancer. 2024. Vol. 5, No. 2. P. 43-52

4.0

https://doi.org/10.37748/2686-9039-2024-5-2-5

https://elibrary.ru/nusqxr

South Russian

Journal of Cancer

ORIGINAL ARTICLE

Южно-Российский

онкологический журнал

Vol. 5

Dynamics of saliva cytokine levels during intraoperative

No. 2, 2024

photodynamic therapy in patients with locally advanced

oral cancer

E. Yu. Komarova1, E. Yu. Zlatnik1, E. F. Komarova1,2, I. A. Novikova1, E. A. Dzhenkova1, M. A. Engibaryan1, A. B. Sagakyants1, V. V. Pozdnyakova1, L. Ya. Rozenko1

1 National Medical Research Centre for Oncology, Rostov-on-Don, Russian Federation 2 Rostov State Medical University, Rostov-on-Don, Russian Federation lil_liza777@mail.ru

ABSTRACT

Purpose of the study. Assessment of the level of certain cytokines in the saliva of patients with primary locally advanced cancer of the oral mucosa in addition to surgical treatment with intraoperative PDT (IPDT).

Patients and methods. Patients with primary locally advanced cancer of the oral mucosa T3-4aN0-2M0 were divided into 2 groups: the main group (30 patients) underwent radical tumor removal supplemented with IPDT and the control group (30 patients) without addition. IPDT was performed using Latus- T (farah) and a chlorin E6 photosensitizer. Cytokine levels were determined in unstimulated whole saliva the day before, on the 3rd and on the 7th day after the operation by the ELISA multiplex analysis method.

Results. A similar dynamic of the cytokine profile of patients of both groups was shown: on the 3rd day after surgery, the levels of G-CSF, IL-6, MIP-1β increased, and GM-CSF and IFN-γ decreased compared with baseline values. On the 7th day, the dynamics of G-CSF, GM-CSF, IL-6 persisted, while IL-8, IL-10, IL-12 changed to the opposite.

Intergroup differences were revealed in the level of IL-1β – on day 3, an increase in the main group and a decrease in the control group. The level of IL-7 on day 7 decreased sharply in the control group and increased statistically significantly in patients receiving IPDT. The main group showed a 4.8-fold increase in IL-8 on day 3 and its 3.6-fold drop on day 7 with the opposite dynamics in the control group. The TNF-α level increased only in the main group on day 7, and in the control group it decreased by 3 and recovered on day 7. On day 3, the MCP-1 level increased in the main group and decreased in the control group. The level of IL-17 in the main group increased on the 3rd day with a further decrease below the baseline, and in the control group it decreased on the 3rd day, followed by a recovery on the 7th. An increase in IL-5 and IL-13 levels on day 3 was noted only in the control group, however, the level of IL-5 in both study periods in the main group was lower than in the control group.

Conclusion. IPDT in patients with primary locally advanced oral cancer causes changes in the cytokine composition of saliva during the first week after surgery, some of which can be associated with an elongation of the relapse-free period in such patients.

Keywords: intraoperative photodynamic therapy, cytokines, saliva, primary locally advanced oral cancer For citation: Komarova E. Yu., Zlatnik E. Yu., Komarova E. F., Novikova I. A., Dzhenkova E. A., Engibaryan M. A., Sagakyants A. B., Pozdnyakova V. V., Rozenko L. Ya. Dynamics of saliva cytokine levels during intraoperative photodynamic therapy in patients with locally advanced oral cancer. South Russian Journal of Cancer. 2024; 5(2):43-52. (In Russ.). https://doi.org/10.37748/2686-9039-2024-5-2-5, https://elibrary.ru/nusqxr For correspondence: Elizaveta Yu. Komarova – PhD student National Medical Research Centre for Oncology, Rostov-on-Don, Russian Federation Address: 63 14 line str., Rostov-on-Don 344037, Russian Federation E-mail: lil_liza777@mail.ru

ORCID: https://orcid.org/0000-0002-1252-8094

SPIN: 9372-1954, AuthorID: 1198314

Scopus Author ID: 57217081452

Compliance with ethical standards: the work followed the ethical principles set forth by the World Medical Association Declaration of Helsinki, 1964, ed. 2013. The study was approved by the Ethics Committee of the National Medical Research Center of Oncology (extract from the protocol of the meeting No. 15 dated 10/12/2021). Informed consent was received from all participants of the study Funding: this work was not funded

Conflict of interest: Zlatnik E. Yu., Dzhenkova E. A., Engibaryan M. A. have been the member of the editorial board of the South Russian Journal of Cancer since 2019, however he has no relation to the decision made upon publishing this article. The article has passed the review procedure accepted in the journal. The authors did not declare any other conflicts of interest The article was submitted 16.11.2023; approved after reviewing 08.02.2024; accepted for publication 09.05.2024

© Komarova E. Yu., Zlatnik E. Yu., Komarova E. F., Novikova I. A., Dzhenkova E. A., Engibaryan M. A., Sagakyants A. B., Pozdnyakova V. V., Rozenko L. Ya., 2024

43

Южно-Российский онкологический журнал. 2024. Т. 5, № 2. С. 43-52

https://doi.org/10.37748/2686-9039-2024-5-2-5

https://elibrary.ru/nusqxr

3.1.6. Онкология, лучевая терапия

ОРИГИНАЛЬНАЯ СТАТЬЯ

Динамика уровней цитокинов слюны при проведении интраоперационной

фотодинамической терапии у больных местно-распространенным раком полости рта

Е. Ю. Комарова1, Е. Ю. Златник1, Е. Ф. Комарова1,2, И. А. Новикова1, Е. А. Дженкова1, М. А. Енгибарян1, А. Б. Сагакянц1, В. В. Позднякова1, Л. Я. Розенко1

1 ФГБУ «Национальный медицинский исследовательский центр онкологии» Министерства здравоохранения Российской Федерации, г. Ростов-на-Дону, Российская Федерация

2 ФГБОУ ВО «Ростовский государственный медицинский университет» Министерства здравоохранения Российской Федерации, г. Ростов-на-Дону, Российская Федерация

lil_liza777@mail.ru

РЕЗЮМЕ

Цель исследования. Оценка уровня некоторых цитокинов в слюне больных первичным местно- распространенным

раком слизистой оболочки полости рта при дополнении хирургического лечения интраоперационной ФДТ (ИФДТ).

Пациенты и методы. Пациенты первичным местно- распространенным раком слизистой оболочки полости рта

T3 -4аN0-2M0 были распределены в 2 группы: основная группа (30 больных) – проведено радикальное удаление

опухоли, дополненное ИФДТ и контрольная группа (30 пациентов) – без дополнения. ИФДТ проводили с помощью

«Латус- Т» (фара) и фотосенсибизатором хлорин Е6. Уровни цитокинов определяли в нестимулированной цельной

слюне за сутки до, на 3-и и на 7-е сутки после проведения операции методом ИФА мультиплекс- анализа.

Результаты. Показана сходная динамика цитокинового профиля больных обеих групп: на 3-и сутки после операции

уровни G-CSF, IL-6, MIP-1β повышались, а GМ-CSF и IFN-γ снижались по сравнению с исходными показателями. На 7-е

сутки характер динамики G-CSF, GМ-CSF, IL-6 сохранялся, а IL-8, IL-10, IL-12 менялся на противоположный.

Межгрупповые различия выявлены по уровню IL-1β – на 3-и сутки повышение в основной и снижение в контрольной

группе. Уровень IL-7 на 7-е сутки резко снижался в контрольной группе и статистически значимо повышался у больных, получавших ИФДТ. В основной группе показано 4,8-кратное повышение IL-8 на 3-и сутки и его 3,6-кратное падение на

7-е с противоположной динамикой в контрольной. Уровень TNF-α возрастал только в основной группе на 7-е сутки, а в контрольной отмечено его снижение на 3-и и восстановление на 7-е сутки. На 3-и сутки уровень MCP-1 возрастал

в основной и снижался в контрольной группе. Уровень IL-17 в основной группе нарастал на 3-и сутки с дальнейшим

снижением ниже исходного, а в контрольной группе снижался на 3-и сутки с последующим восстановлением на 7-е.

Нарастание уровней IL-5 и IL-13 на 3-и сутки отмечены только в контрольной группе, однако, уровень IL-5 в оба срока

исследования в основной группе был ниже, чем в контрольной.

Заключение. ИФДТ у больных первичным местно- распространенным раком полости рта вызывает изменения цитокинового состава слюны в течение первой недели после операции, часть из которых можно связать с удлинением

безрецидивного периода у таких пациентов.

Ключевые слова: интраоперационная фотодинамическая терапия, цитокины, слюна, первичный

местно-распространенный рак полости рта

Для цитирования: Комарова Е. Ю., Златник Е. Ю., Комарова Е. Ф., Новикова И. А., Дженкова Е. А., Енгибарян М. А., Сагакянц А. Б., Позднякова В. В., Розенко Л. Я. Динамика уровней цитокинов слюны при проведении интраоперационной фотодинамической терапии

у больных местно- распространенным раком полости рта. Южно- Российский онкологический журнал. 2024; 5(2):43-52.

https://doi.org/10.37748/2686-9039-2024-5-2-5, https://elibrary.ru/nusqxr Для корреспонденции: Комарова Елизавета Юрьевна – аспирант, ФГБУ «Национальный медицинский исследовательский центр

онкологии» Министерства здравоохранения Российской Федерации, г. Ростов-на- Дону, Российская Федерация

Адрес: 344037, Российская Федерация, г. Ростов-на- Дону, ул. 14-я линия, д. 63

E-mail: lil_liza777@mail.ru

ORCID: https://orcid.org/0000-0002-1252-8094

SPIN: 9372-1954, AuthorID: 1198314

Scopus Author ID: 57217081452

Соблюдение этических стандартов: в работе соблюдались этические принципы, предъявляемые Хельсинкской декларацией Всемирной

медицинской ассоциации (World Medical Association Declaration of Helsinki, 1964, ред. 2013). Исследование одобрено этическим

комитетом ФГБУ «Национальный медицинский исследовательский центр онкологии» Министерства здравоохранения Российской

Федерации (выписка из протокола заседания № 15 от 12.10.2021 г.). Информированное согласие получено от всех участников

исследования

Финансирование: финансирование данной работы не проводилось

Конфликт интересов: Златник Е. Ю., Дженкова Е. А., Енгибарян М. А. являются членами редакционной коллегии журнала «Южно-Российский онкологический журнал» с 2019 г., но не имеют никакого отношения к решению опубликовать эту статью. Статья прошла

принятую в журнале процедуру рецензирования. Об иных конфликтах интересов авторы не заявляли

Статья поступила в редакцию 16.11.2023; одобрена после рецензирования 08.02.2024; принята к публикации 09.05.2024

44

Южно-Российский онкологический журнал 2024. Т. 5, № 2. С. 43-52

Комарова Е. Ю., Златник Е. Ю., Комарова Е. Ф., Новикова И. А., Дженкова Е. А., Енгибарян М. А., Сагакянц А. Б., Позднякова В. В., Розенко Л. Я. Динамика

уровней цитокинов слюны при проведении интраоперационной фотодинамической терапии у больных местно- распространенным раком полости рта

primary locally advanced cancer of the oral mucosa

INTRODUCTION

in addition to surgical treatment with intraoperative

PDT (IPDT).

Currently, it is recognized that under the influence

of inflammation, neoplastic and stromal cells interact

PATIENTS AND METHODS

and control the evolution of the tumor, producing

cytokines that modulate the antitumor immune re-

The study group included 60 patients with primary

sponse [1–3]. The similarity of the cytokine pattern

locally advanced cancer of the oral mucosa who were

for tumors of different localization and histogen-

being treated by the Department of Head and Neck

esis has been demonstrated [4], however, there is

Tumors of the Federal State Budgetary Institution

no consensus on whether cytokines play a decisive

"National Medical Research Center of Oncology" of pro-oncogenic or anti-oncogenic role [5]. Both can be

the Ministry of Health of the Russian Federation. The

assumed based on the known biological properties

average age of the patients was 63.1 ± 14.3 years.

of cytokines as stimulants, on the one hand, prolifer-

The distribution of patients by gender was as follows:

ation and neo angiogenesis, and on the other hand,

male – 48 people, female – 12 people. In all patients,

the immune response.

the diagnosis was morphologically verified and cor-

In recent decades, the participation of various

responded to stages III–IV (T3-4aN0-2M0) of cancer.

pro- or anti-inflammatory cytokines of the tumor

According to the morphological structure, the tumors

microenvironment of squamous cell carcinoma of

were represented by squamous cell carcinoma, of

the oral cavity in the oncogenesis of a tumor of this

which moderate–differentiated – 79 %, highly differ-

localization has been shown [6–8]. Most studies

entiated – 16 % and low–differentiated – 5 %.

describe elevated levels of IL-1β, 6, 8, 10, TNF-α in

The patients were randomized into 2 groups:

patients with oral cancer compared with healthy

the main group (30 patients) – patients who un-

ones at both the local and systemic levels [9, 10].

derwent radical tumor removal supplemented with

Moreover, a higher content of IL-1β and IL-6 was

IPDT and the control group (30 patients) – with-

demonstrated in tumors of the oral mucosa than

out the supplement. The patients in the groups

in areas of unchanged mucosa [11]. Literature data

were comparable in age, gender and location of

show that hyperproduction of cytokines by tumor

the primary focus. All patients underwent surgical

cells and its microenvironment in patients with oral

intervention in the volume of radical removal of the

cancer is one of the causes of tumor spread.

tumor of the tongue and oral mucosa with cervical

Thus, when developing approaches to the treat-

lymphadenectomy in a volume adequate to the

ment of oral cancer, it is important to analyze the

prevalence of the tumor process, on the affected

local and systemic cytokine status of such patients,

side or bilateral (with damage to the anterior parts

as well as the induction of an antitumor immune

of the oral cavity).

response. The latter can be achieved with photo-

The study was approved by the Ethics Committee

dynamic therapy (PDT), one of the mechanisms

of the National Medical Research Center of Oncol-

of antitumor effectiveness of which is immuno-

ogy Protocol No. 15 dated 10/12/2021, all patients

genic cell death [12]. PDT induces it through the

signed a voluntary informed consent.

release of damage- associated molecular patterns

Intraoperative PDT was performed in accordance

(DAMPs) [13]. PDT-induced acute inflammation and

with the developed and patented method (RF Pat-

immunogenic cell death are considered the initial

ent No. 2797433) [15]. At the first stage, patients

step in the implementation of its immunostimulat-

with locally advanced cancer of the mucous mem-

ing effect [14], which manifests itself in a tumor-

brane of the oral cavity and tongue underwent sur-

specific immune response [15]. The role and place

gical treatment, and then, after hemostasis of the

of immune mechanisms in the realization of the

postoperative wound of the oral cavity, PDT was

antitumor effect of PDT are summarized in a recent

performed after covering the healthy surrounding

review [14].

tissues around the surgical field with sterile eight-

The purpose of the study was to assess the level

layer gauze wipes. Latus- T (headlight) was used for

of certain cytokines in the saliva of patients with

the session, with parameters: wavelength 662 nm,

45

South Russian Journal of Cancer 2024. Vol. 5, No. 2. P. 43-52

Komarova E. Yu., Zlatnik E. Yu., Komarova E. F., Novikova I. A., Dzhenkova E. A., Engibaryan M. A., Sagakyants A. B., Pozdnyakova V. V., Rozenko L. Ya. Dynamics of saliva cytokine levels during intraoperative photodynamic therapy in patients with locally advanced oral cancer power density 45 MW, light energy 200–300 J/cm2.

compared with baseline values; the content of the

The lamp was placed in such a way that the area

remaining cytokines was multidirectional in the

of laser light emission covered both the bed of the

compared groups of patients.Some of the indicators

removed tumor and the edges of the surgical wound.

changed statistically significantly in only one of the

The duration of exposure was calculated depending

groups, which is apparently related to the effect of

on the size of the bed of the removed tumor of the

PDT (Fig. 1B, 1G).

oral cavity according to the formula: T = T0 × nw/ kp,

On the 7th day, the dynamics of G-CSF, GM-CSF,

where T0 is the tabular value of the irradiation time,

IL-6 persisted, IL-8, IL-10, IL-12 changed to the op-

nw is a coefficient showing how many times the

posite, the levels of other cytokines changed in the

energy density WS (J/ cm2) to be collected by the

main and control groups in different ways (Fig. 2A-D).

surface differs from the tabular WS/0 = 100 J/ cm2:

Differences were revealed between the main and

nw = WS/100, kp is a coefficient showing how

control groups in terms of colony- stimulating factors:

many times the laser power differs from the tabular

an increase in G-CSF and a decrease in GM-CSF on

P0 = 100 MW: kp = P/100. After completion of the

both the 3rd and 7th days after surgery compared

photodynamic therapy stage, plastic surgery of the

with baseline values, and in the main group, chang-

postoperative defect was performed. Chloride E6

es in G-CSF are less pronounced than in the control

was used as a photosensitizer, which was adminis-

group, and in both The duration of the study was at

tered intravenously in a single dose of 2 mg per 1 kg

lower values (Fig. 1A, 2A). So, in the control group,

of patient weight 3–3.5 hours before the expected

the level of G-CSF increased 22.8 times on the 3rd

end time of tumor removal. The immediate results

day after surgery, and 13 times in the main group,

of complex treatment supplemented with IPDT were

and on the 7th day it was 22.2 and 4.8 times higher

evaluated in patients with locally advanced oral mu-

than the initial one, respectively, i. e. the decrease on

cosal cancer.

the 7th day after the initial increase by 3 occurred

Cytokine levels (G-CSF, GM-CSF, IFN-γ, IL-1β, IL-2,

only after the effect of PDT.

IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17,

IL-1β levels on day 3 increased in the main

MCP-1, MIP-1β, TNF-α) were determined in saliva

group from 410.1 [321.9; 522.6] to 550.8 [528.7;

by ELISA multiplex analysis (Bio- Plex Pro Human

590.6] pg/ ml and decreased in the control group to

Cytokine Assays 17-Plex Panel, Bio- Rad, USA). The

102.1 [22.1; 159] pg/ml, and on the 7th the reverse

collection of unstimulated whole saliva for the study

pattern was observed (Fig. 1B, 2B).

was carried out from 8 to 9 a. m. the day before the

There was a 3.3-fold increase in IL-6 in the sali-

operation, on the 3rd and on the 7th day after it. The

va of patients of both groups on day 3 compared

results were expressed in pg/ml.

with the baseline, which continued on day 7 with

Statistical analysis of the study results was carried

higher rates in the main group (1856.2 [1753.1;1975]

out using the Statistica 12.0 program (StatSoft Inc.,

versus 1356.9 [1261.3; 1450.7] pg/ml, p < 0.05), USA), MedCalc (version 9.3.5.0). Since the distribution

(Fig. 1A, 2A).

was not normal, the Mann- Whitney criterion was used

The dynamics of IL-7 consisted in an increase in

to compare intergroup indicators, and the Wilcoxon

the level of this cytokine in the saliva of patients of

criterion was used to compare indicators in dynamics;

both groups on the 3rd day after surgery, statisti-

the differences were considered statistically signifi-

cally significant only in the control group (by 53 %)

cant at p < 0.05. The data are presented as a median and multidirectional on the 7th day – a sharp drop

with upper and lower quartiles (Me [LQ, UQ]).

(to 0) in the control group and a continued increase

that reached statistical significance (15.3 [13.6; 17.2]

STUDY RESULTS AND DISCUSSION

versus 10.2 [6.8; 13.5] before treatment; p < 0.05) in patients receiving IPDT.

The results of the study are shown in Fig. 1A-1G,

In the main group, there was a pronounced

2A-2G.

(4.8-fold) transient increase in IL-8 content on day

On the 3rd day after surgery in patients of both

3 and its 3.6-fold drop on day 7; in the control group,

groups, the levels of G-CSF, IL-6, MIP-1β increased

on the contrary, it decreased on day 3 and increased

(Fig. 1A), and GM-CSF and IFN-γ decreased (Fig. 1B)

on day 7, exceeding the indicator of the main group

46

Южно-Российский онкологический журнал 2024. Т. 5, № 2. С. 43-52

Комарова Е. Ю., Златник Е. Ю., Комарова Е. Ф., Новикова И. А., Дженкова Е. А., Енгибарян М. А., Сагакянц А. Б., Позднякова В. В., Розенко Л. Я. Динамика

уровней цитокинов слюны при проведении интраоперационной фотодинамической терапии у больных местно- распространенным раком полости рта

in the last period (Me 2883.1 [2621.8;3338.2] and

dynamics in both groups, increasing by 3 and re-

2006.4 [1934.3;2310.4] pg/ml, respectively, p < 0.05), turning to the baseline on the 7th day after surgery

(Fig. 1A, 2B). The TNF-α level increased only in the

(Fig. 1A, 2G).

main group on day 7, and in the control group it de-

There was a decrease in IFN-γ and IL-4, expressed

creased by 3 and recovered on day 7 (Fig. 1D, 2D).

in the main group only on the 7th day with indicators

On the 3rd day, there was a multidirectional dynam-

at this time lower than the control ones (Fig. 2B).

ics of MCP-1 levels: an increase in the main group

The median level of IFN-γ in the main group on

and a decrease in the control group (Fig. 1D), and

day 7 was 4.7 [4.2; 5.2] pg/ml, and in the control

on the 7th in both groups, the indicators become

group 6.8 [6.3; 7.4] pg/ml versus 10.6 [8.4; 12.4]

3.5 and 4.6 times lower, respectively, than the initial

pg/ ml before treatment; for IL-4, the indicators were

one (Fig. 2D). The content of MIP-1β has a similar

4.1 [3.2; 4.7] and 8,6 [7,2; 8,9] versus 10,9 [8,7; 14,2]

values 2500

values

40

e

e

ativ 2000

ativ

20

eoper 1500

eoper

0

pr

pr

om

om

fr 1000

fr

- 20

ences

500

ences

- 40

er

er

0

- 60

% of diff -500

% of diff

- 80

G-CSE

IL-6

IL-8

IL-13

MIP-1β

GM-CSF

IFN-γ

IL-1β

IL-4

Main group, 3rd day

А

Main group, 3rd day

B

Control group, 3rd day

Control group, 3rd day

30

values

60

values

20

e

e

10

ativ

50

ativ

0

eoper

40

eoper

pr

pr

-10

om

30

om

-20

fr

fr

-30

ences

20

ences

-40

er

er

-50

10

-60

% of diff

% of diff

0

-70

IL-5

IL-7

IL-10

IL-12

IL-2

IL-17

MCP-1

TNF-α

Main group, 3rd day

Main group, 3rd day

C

D

Control group, 3rd day

Control group, 3rd day

Fig. 1. The concentration of cytokines in the saliva of patients on the 3rd postoperative day.

Note: * – statistically significant differences compared to the level before surgery, p < 0.05; ** – statistically significant differences compared to the level of the control group, p < 0.05

47

South Russian Journal of Cancer 2024. Vol. 5, No. 2. P. 43-52

Komarova E. Yu., Zlatnik E. Yu., Komarova E. F., Novikova I. A., Dzhenkova E. A., Engibaryan M. A., Sagakyants A. B., Pozdnyakova V. V., Rozenko L. Ya. Dynamics of saliva cytokine levels during intraoperative photodynamic therapy in patients with locally advanced oral cancer pg/ml, respectively; for both cytokines, the differenc-14.7 [13.5; 16.7] pg/ml before treatment; p < 0.05) es between the indicators of the main and control

only for the control group), which can be regarded

groups are statistically significant ( p < 0.05). The positively, given the role of this cytokine in the po-dynamics of IL-2 levels in the form of an increase

larization of T-helper cells in Th1, providing cellular

was detected only in the control group on day

immunity.

7 (Me 29.6 [26.9; 30.7] versus 21.4 [17.4; 25.5] pg/ ml

IL-17 levels demonstrate different phases of

before treatment, p < 0.05), (Fig. 2B).

changes: in the main group, a transient increase on

A decrease in IL-12 levels occurs in both groups

the 3rd day with a further decrease below the initial

on day 7 (Fig. 2B), and it is less pronounced in the

one, and in the control group, a decrease on the 3rd

main group than in the control group (Me 11.7 [10.6;

day followed by a recovery on the 7th (Fig. 1G, 2G).

12.8] and 7.9 [7.2; 9.3], respectively, versus

The increase in the levels of cytokines polarizing

100

values 2500

values

80

e

e

60

ativ

ativ

2000

40

eoper

eoper

20

pr

pr

1500

0

om

om

fr

fr

-20

1000

-20

ences

ences

-40

er

er

500

-60

-80

% of diff

0

% of diff -100

G-CSF

IL-6

IFN-γ

IL-1β

IL-4

IL-7

IL-8

IL-10

Main group, 7th day

A

Main group, 7th day

Б

Control group, 7th day

Control group, 7th day

40

30

values

values

20

e

30

e

10

ativ

20

ativ

0

eoper

10

eoper

-10

pr

0

pr

-20

om fr

om

-10

fr

-30

-20

-40

ences

ences

er

-50

-30

er

-60

-40

-70

% of diff

-50

% of diff

-80

GM-CSF

IL-2

IL-5

IL-12

IL-13

IL-17

MCP-1

MIP-1β

TNF-α

Main group, 7th day

В

Main group, 7th day

Г

Control group, 7th day

Control group, 7th day

Fig. 2. The concentration of cytokines in the saliva of patients on the 7th postoperative day.

Note: * – statistically significant differences compared to the level before surgery, p < 0.05; ** – statistically significant differences compared to the levels in the control group, p < 0.05

48

Южно-Российский онкологический журнал 2024. Т. 5, № 2. С. 43-52

Комарова Е. Ю., Златник Е. Ю., Комарова Е. Ф., Новикова И. А., Дженкова Е. А., Енгибарян М. А., Сагакянц А. Б., Позднякова В. В., Розенко Л. Я. Динамика

уровней цитокинов слюны при проведении интраоперационной фотодинамической терапии у больных местно- распространенным раком полости рта

CD4+ in Th2, IL-5 and IL-13 on day 3, noted only in

the importance of TNF-α, IL-6, IL-1β, as well as

the control group, is also transient. However, the

heat shock proteins, metabolites of arachidon-

level of IL-5 in both study periods in the main group

ic acid [17]. In this regard, cytokines that have

was lower than in the control group (Fig. 1B, 2b):

a pro-inflammatory effect, in particular, the ability

on day 3, 136.3 [130.2; 144.4] versus 173.6 [168.7;

to increase vascular permeability, and are usually

176.8] pg/ml, on day 7, 117.4 [106.6; 125.2] versus

considered pro-oncogenic due to their stimulating

138.5 [131.7; 147.2] pg/ml, respectively, at both pe-

effect on neoangiogenesis, have a positive value

riods p < 0.05), which, taking into account the data in PDT for the realization of its effect. The com-described in the literature on its association with the

parative dynamics of saliva cytokines obtained

recurrence of some tumors [16], is probably one of

by us in patients of the main and control groups

the positive aspects of the action of IPDT.

suggests earlier peaks in IL-1β and IL-8 concen-

The importance of chronic inflammation and

trations occurring on the 3rd day after intraoper-

its accompanying cytokine production in tumor

ative PDT and high local levels of TNF-α and IL-6,

growth is known. The pro-oncogenic role of most

which persist throughout the follow-up period.

known cytokines in neoplasia has been repeat-

On the contrary, anti-inflammatory cytokines, for

edly described, realized through the activation

example, IL-10 and TGF-β, suppress the effects of

of various transcription factors and signaling

PDT [18]. It is known about the importance of IL-7

pathways, stimulation of proliferation, neoangio-

in inflammation that its local content increases

genesis, epithelial- mesenchymal transition, and

due to the production of macrophages, dendritic

other processes [4]. However, it seems difficult

cells, fibroblasts, and its target are T lymphocytes

to distinguish the importance of cytokines for

expressing a receptor for it. IL-7 stimulates main-

malignant growth and for the immune response,

ly Th1 and Th17 lymphocytes mediating cellular

of which they are an integral component, as a re-

immunity [19]. A recent review on the role of IL-7

sult of which their antitumor activity is carried

in tumors indicated that it inhibits the growth of

out. Probably, a lot depends on their number, ra-

melanoma, enhances the action of IFN-γ, restores

tio, expression of receptors to them, etc. Unlike

the activity of CD8+ T lymphocytes by reducing

chronic, acute inflammation, moreover, is not mi-

their expression of PD-1 [20]. Based on this, it can

crobial, but induced by a physical factor such as

be assumed that the decrease in the level of this

PDT, apparently exhibits an inhibitory effect on

cytokine observed by us after a transient increase

tumor growth both directly by destroying its cells

in the saliva of patients in the control group is

and indirectly through the immune system [15].

prognostically unfavorable for the state of local

In this regard, PDT-induced hyperproduction of

cellular immunity, and, on the contrary, its stable

cytokines in the lesion, especially after removal

increase in the saliva of patients receiving IPDT

of the tumor, not only contributes to its sanitation,

reflects stimulation of the T-cell link.

accelerating wound cleansing and healing, but

The role of granulocytes and related cytokines

also may have positive long-term consequences

in the body of the tumor carrier is twofold. There

associated with an increase in the recurrence-free

are numerous literature data on the pro-oncogenic

period due to the destruction of residual tumor

effect of neutrophils [21], as well as chemokines

cells and induction of an immune response to

that attract them to the focus. Neutrophilic "traps"

DAPMs. Our results partially confirm this. Oxi-

promote the survival of circulating tumor cells in the

dative stress caused by PDT stimulates the re-

blood and tumor metastasis [22, 23]. Nevertheless,

lease of proinflammatory cytokines into the tu-

neutrophils, as participants in the generation of

mor microenvironment, which also affects their

ROS, are given important importance in the action

content in saliva. At the beginning of PDT-induced

of PDT [24]. However, since G-CSF promotes the

inflammation, tumor vessels become permeable

release of immature granulocytes with immunosup-

to adhesion proteins (ICAM 1, VCAM 1, selectins),

pressive effects (MDSC) from the bone marrow into

thus contributing to massive infiltration of the

the peripheral blood and tissues, we consider a low-

tumor by immune cells producing a wide range

er level of it in our patients receiving IPDT compared

of cytokines, of which the literature emphasizes

with the control group as a positive moment.

49

South Russian Journal of Cancer 2024. Vol. 5, No. 2. P. 43-52

Komarova E. Yu., Zlatnik E. Yu., Komarova E. F., Novikova I. A., Dzhenkova E. A., Engibaryan M. A., Sagakyants A. B., Pozdnyakova V. V., Rozenko L. Ya. Dynamics of saliva cytokine levels during intraoperative photodynamic therapy in patients with locally advanced oral cancer CONCLUSION

with the resulting clinical effect, consisting in pro-

longation of the relapse-free period in such patients

Intraoperative use of PDT in patients with locally

(RF Patent No. 2797433). However, further studies

advanced oral cancer causes changes in the cytokine

are required to clarify the role of cytokines in the

composition of saliva that develop during the first

antitumor effectiveness of the applied photodynamic

week after surgery, some of which can be associated

effect in patients with locally advanced oral cancer.

References

1. Zlatnik EYu, Sagakyants AB, Shulgina OG, Shevchenko AN, Filatova EV, Belyakova LI, et al. Local cytokine levels as prognostic factors for early relapse of non-muscle-invasive bladder carcinoma. Medical Immunology. 2023;25(5):1135–1140.

(Russ.). https://doi.org/10.15789/1563-0625-LCL-2723, EDN: JEPHWY

2. Rybkina VL, Adamova GV, Oslina DS. The role of cytokines in the pathogenesis of malignant neoplasms. Siberian Scientific Medical Journal. 2023;43(2):15–28. (In Russ.). https://doi.org/10.18699/SSMJ20230202, EDN: MRRKJB

3. Kartikasari AER, Huertas CS, Mitchell A, Plebanski M. Tumor-Induced Inflammatory Cytokines and the Emerging Diagnos-tic Devices for Cancer Detection and Prognosis. Front Oncol. 2021;11:692142. https://doi.org/10.3389/fonc.2021.692142

4. Lan T, Chen L, Wei X. Inflammatory Cytokines in Cancer: Comprehensive Understanding and Clinical Progress in Gene Therapy. Cells. 2021 Jan 8;10(1):100. https://doi.org/10.3390/cells10010100

5. Aghayev T, Titerina EK, Khoreva MV, Gankovskaya LV. Role of cytokines in hepatocellular carcinoma. Medical Immunology.

2022;24(5):889–902. (In Russ.). https://doi.org/10.15789/1563-0625-ROC-2512, EDN: FXTNRU

6. Ralli M, Grasso M, Gilardi A, Ceccanti M, Messina MP, Tirassa P, et al. The role of cytokines in head and neck squamous cell carcinoma: A review. Clin Ter. 2020;171(3):e268–e274. https://doi.org/10.7417/CT.2020.2225

7. Caruntu A, Scheau C, Codrici E, Popescu ID, Calenic B, Caruntu C, et al. The Assessment of Serum Cytokines in Oral Squamous Cell Carcinoma Patients: An Observational Prospective Controlled Study. J Clin Med. 2022 Sep 14;11(18):5398.

https://doi.org/10.3390/jcm11185398

8. Chadwick JW, Macdonald R, Ali AA, Glogauer M, Magalhaes MA. TNFα Signaling Is Increased in Progressing Oral Potentially Malignant Disorders and Regulates Malignant Transformation in an Oral Carcinogenesis Model. Front Oncol.

2021;11:741013. https://doi.org/10.3389/fonc.2021.741013

9. Dikova V, Jantus-Lewintre E, Bagan J. Potential Non-Invasive Biomarkers for Early Diagnosis of Oral Squamous Cell Carcinoma. J Clin Med. 2021 Apr 13;10(8):1658. https://doi.org/10.3390/jcm10081658

10. Val M, Sidoti Pinto GA, Manini L, Gandolfo S, Pentenero M. Variations of salivary concentration of cytokines and chemokines in presence of oral squamous cell carcinoma. A case-crossover longitudinal prospective study. Cytokine. 2019

Aug;120:62–65. https://doi.org/10.1016/j.cyto.2019.04.009

11. Zlatnik EYu, Svetitskii PV, Novikova IA, Nistratov GP, Aedinova IV, Volkova VL et al. Factors of local immunity in patients with oral mucosa and tongue carcinoma with absence and presence of regional metastases and recurrences. Basic Research. 2015:1-5;953–957. EDN: TWTPFJ

12. Falk-Mahapatra R, Gollnick SO. Photodynamic Therapy and Immunity: An Update. Photochem Photobiol. 2020

May;96(3):550–559. https://doi.org/10.1111/php.13253

13. Donohoe C, Senge MO, Arnaut LG, Gomes-da-Silva LC. Cell death in photodynamic therapy: From oxidative stress to anti-tumor immunity. Biochim Biophys Acta Rev Cancer. 2019 Dec;1872(2):188308. https://doi.org/10.1016/j.bbcan.2019.07.003

14. Huis In ’t Veld RV, Heuts J, Ma S, Cruz LJ, Ossendorp FA, Jager MJ. Current Challenges and Opportunities of Photodynamic Therapy against Cancer. Pharmaceutics. 2023 Jan 18;15(2):330. https://doi.org/10.3390/pharmaceutics15020330

15. RF Patent No. 2797433 Russian Federation, IPC A61B 18/20 (2006.1), A61N5/067 (2006.01), A61K 6/00 (2006.01), A61P

35/00 (2006.01) Method of intraoperative photodynamic therapy in the combined treatment of primary locally advanced 50

Южно-Российский онкологический журнал 2024. Т. 5, № 2. С. 43-52

Комарова Е. Ю., Златник Е. Ю., Комарова Е. Ф., Новикова И. А., Дженкова Е. А., Енгибарян М. А., Сагакянц А. Б., Позднякова В. В., Розенко Л. Я. Динамика

уровней цитокинов слюны при проведении интраоперационной фотодинамической терапии у больных местно- распространенным раком полости рта

tongue cancer: N 2022118036: application 07/01/2022: published 06/05/2023. Kit OI, Komarova EYu, Yengibaryan MA, Volkova VL, Chertova NA, Komarova EF, et al; applicant of the National Medical Research Center of Oncology. 10 p. (In Russ.).

16. Kumar P, Ranmale S, Mehta S, Tongaonkar H, Patel V, Singh AK, et al. Immune profile of primary and recurrent epithelial ovarian cancer cases indicates immune suppression, a major cause of progression and relapse of ovarian cancer. J Ovarian Res. 2023 Jun 15;16(1):114. https://doi.org/10.1186/s13048-023-01192-4

17. Maeding N, Verwanger T, Krammer B. Boosting Tumor-Specific Immunity Using PDT. Cancers (Basel). 2016 Oct 6;8(10):91.

https://doi.org/10.3390/cancers8100091

18. Reginato E, Wolf P, Hamblin MR. Immune response after photodynamic therapy increases anti-cancer and anti-bacterial effects. World J Immunol. 2014 Mar 27;4(1):1–11. https://doi.org/10.5411/wji.v4.i1.1

19. Bikker A, Hack CE, Lafeber FPJG, van Roon JAG. Interleukin-7: a key mediator in T cell-driven autoimmunity, inflammation, and tissue destruction. Curr Pharm Des. 2012;18(16):2347–2356. https://doi.org/10.2174/138161212800165979

20. Wang C, Kong L, Kim S, Lee S, Oh S, Jo S, et al. The Role of IL-7 and IL-7R in Cancer Pathophysiology and Immunotherapy.

Int J Mol Sci. 2022 Sep 8;23(18):10412. https://doi.org/10.3390/ijms231810412

21. Szczerba BM, Castro-Giner F, Vetter M, Krol I, Gkountela S, Landin J, et al. Neutrophils escort circulating tumour cells to enable cell cycle progression. Nature. 2019 Feb;566(7745):553–557. https://doi.org/10.1038/s41586-019-0915-y 22. Vorobjeva NV, Chernyak BV. NETosis: Molecular Mechanisms, Role in Physiology and Pathology. Biochemistry (Mosc).

2020 Oct;85(10):1178–1190. https://doi.org/10.1134/S0006297920100065

23. Azzouz D, Khan MA, Palaniyar N. ROS induces NETosis by oxidizing DNA and initiating DNA repair. Cell Death Discov. 2021

May 18;7(1):113. https://doi.org/10.1038/s41420-021-00491-3

24. Brackett CM, Muhitch JB, Evans SS, Gollnick SO. IL-17 promotes neutrophil entry into tumor-draining lymph nodes following induction of sterile inflammation. J Immunol. 2013 Oct 15;191(8):4348–4357. https://doi.org/10.4049/jimmunol.1103621

Information about authors:

Elizaveta Yu. Komarova – PhD student, National Medical Research Centre for Oncology, Rostov-on-Don, Russian Federation ORCID: https://orcid.org/0000-0002-1252-8094, SPIN: 9372-1954, AuthorID: 1198314, Scopus Author ID: 57217081452

Elena Yu. Zlatnik – Dr. Sci. (Med.), professor, chief researcher of the Laboratory of Tumor Immunophenotyping, National Medical Research Centre of Oncology, Rostov-on-Don, Russian Federation

ORCID: https://orcid.org/0000-0002-1410-122X, SPIN: 4137-7410, AuthorID: 327457, ResearcherID: AAI-1311-2020, Scopus Author ID: 6603160432

Ekaterina F. Komarova – Dr. Sci. (Biol.), Professor of the RAS, Leading Researcher, National Medical Research Centre for Oncology, Rostov-on-Don, Russian Federation; Dean of the Faculty of Clinical Psychology, Head of the Department of Biomedicine (and Psychophysiology), Rostov State Medical University, Rostov-on-Don, Russian Federation

ORCID: https://orcid.org/0000-0002-7553-6550, SPIN: 1094-3139, AuthorID: 348709, ResearcherID: T-4520-2019, Scopus Author ID: 55890096600

Inna A. Novikova – Dr. Sci. (Med.), deputy director for science, National Medical Research Centre for Oncology, Rostov-on-Don, Russian Federation ORCID: https://orcid.org/0000-0002-6496-9641, SPIN: 4810-2424, AuthorID: 726229, ResearcherID: E-7710-2018, Scopus Author ID: 7005153343

Elena A. Dzhenkova – Dr. Sci. (Biol.), professor, academic secretary, National Medical Research Centre, Rostov-on-Don, Russian Federation ORCID: https://orcid.org/0000-0002-3561-098X, SPIN: 6206-6222, AuthorID: 697354, ResearcherID: K-9622-2014, Scopus Author ID: 6507889745

Marina A. Engibaryan – Dr. Sci. (Med.), head of Department of Head and Neck Tumors, National Medical Research Centre for Oncology, Rostov-on-Don, Russian Federation

ORCID: https://orcid.org/0000-0001-7293-2358, SPIN: 1764-0276, AuthorID: 318503, Scopus Author ID: 57046075800

Aleksandr B. Sagakyants – Cand. Sci. (Biol.), associate professor, head of the laboratory of Tumor Immunophenotyping, National Medical Research Centre for Oncology, Rostov-on-Don, Russian Federation

ORCID: https://orcid.org/0000-0003-0874-5261, SPIN: 7272-1408, AuthorID: 426904, ResearcherID: M-8378-2019, Scopus Author ID: 24329773900

Viktoriya V. Pozdnyakova – Dr. Sci. (Med.), Professor, Oncologist of the Department of Reconstructive Plastic Surgery and Oncology, National Medical Research Centre for Oncology, Rostov-on-Don, Russian Federation ORCID: https://orcid.org/0000-0002-3782-6899, SPIN: 7306-2034, AuthorID: 700139, ResearcherID: ATT-67-07-2020, Scopus Author ID: 54380529400

Lyudmila Ya. Rozenko – Dr. Sci. (Med.), professor, radiotherapist at the Radiotherapy Department No. 2, National Medical Research Centre for Oncology, Rostov-on-Don, Russian Federation

ORCID: https://orcid.org/0000-0001-7032-8595, SPIN: 8879-2251, AuthorID: 421802, Scopus Author ID: 5397560100

51

South Russian Journal of Cancer 2024. Vol. 5, No. 2. P. 43-52

Komarova E. Yu., Zlatnik E. Yu., Komarova E. F., Novikova I. A., Dzhenkova E. A., Engibaryan M. A., Sagakyants A. B., Pozdnyakova V. V., Rozenko L. Ya. Dynamics of saliva cytokine levels during intraoperative photodynamic therapy in patients with locally advanced oral cancer Contribution of the authors:

Komarova E. Yu. – writing the source text;

Zlatnik E. Yu. – development of methodology, final conclusions; Komarova E. F. – research concept;

Novikova I. A. – scientific guidance, final conclusions;

Dzhenkova E. A. – participation in the development of research design; scientific guidance; Engibaryan M. A. – scientific guidance;

Sagakyants A. B. – development of methodology, revision of the text; Pozdnyakova V. V. – writing the source text;

Rozenko L. Ya. – revision of the text, final conclusions.

52