rpmjResearch'n Practical Medicine JournalResearch and Practical Medicine Journal2410-1893"QUASAR", LLC10.17709/2409-2231-2020-7-1-3rpmj-497Research ArticleОригинальные статьи. Онкология, лучевая терапияOriginal Articles. ОncologyВлияние нокаута по гену урокиназы у мышей C57BL/6-PlautmI. IBugThisPlau6FDhu/ GFDhu на факторы роста при злокачественной меланомеInfluence of urokinase gene-knockout in C57BL/6-PlautmI. IBugThisPlau6FDhu/GFDhu mice on growth factors in malignant melanomahttps://orcid.org/0000-0003-3618-6890ФранциянцЕ. М.FrantsiyantsE. M.

Франциянц Елена Михайловна – д.б.н., профессор, заместитель генерального директора по научной работе, руководитель лаборатории изучения патогенеза злокачественных опухолей 

SPIN: 9427-9928

Elena M. Frantsiyants – Dr. Sci. (Biol.), professor, deputy director general for science, head of the laboratory for the study of the pathogenesis of malignant tumors 

SPIN: 9427-9928

https://orcid.org/0000-0002-3972-2452КаплиеваИ. В.KaplievaI. V.

Каплиева Ирина Викторовна – д.м.н., старший научный сотрудник лаборатории изучения патогенеза злокачественных опухолей 

SPIN: 5047-1541

Irina V. Kaplieva – Dr. Sci. (Med.), senior researcher of the laboratory for the study of pathogenesis of malignant tumors

SPIN: 5047-1541

https://orcid.org/0000-0002-7395-3086НескубинаИ. В.NeskubinaI. V.

Нескубина Ирина Валерьевна – к.б.н., старший научный сотрудник лаборатории изучения патогенеза злокачественных опухолей 

SPIN: 3581-8531

Irina V. Neskubina – Cand. Sci. (Biol.), senior researcher of the laboratory for the study of pathogenesis of malignant tumors 

SPIN: 3581-8531

https://orcid.org/0000-0002-2302-8271БандовкинаВ. А.BandovkinaV. A.

Бандовкина Валерия Ахтямовна – к.б.н., старший научный сотрудник лаборатории изучения патогенеза злокачественных опухолей 

SPIN: 8806-2641

Valeriya A. Bandovkina – Cand. Sci. (Biol.), senior researcher of the laboratory for the study of pathogenesis of malignant tumors 

SPIN: 8806-2641

https://orcid.org/0000-0002-9749-2747ТрепитакиЛ. К.TrepitakiL. K.

Трепитаки Лидия Константиновна – научный сотрудник лаборатории изучения патогенеза злокачественных опухолей 

SPIN: 2052-1248

Lidiya K. Trepitaki – assistant researcher of the laboratory for the study of pathogenesis of malignant tumors

SPIN: 2052-1248

https://orcid.org/0000-0002-4318-7587СуриковаЕ. И.SurikovaE. I.

Сурикова Екатерина Игоревна – к.б.н., старший научный сотрудник лаборатории изучения патогенеза злокачественных опухолей 

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

SPIN: 2401-4115

Ekaterina I. Surikova – Cand. Sci. (Biol.), senior researcher of the laboratory for the study of pathogenesis of malignant tumors 

SPIN: 2401-4115

sunsur2000@mail.ruhttps://orcid.org/0000-0002-3711-8155ЧеряринаН. Д.CheryarinaN. D.

Черярина Наталья Дмитриевна – врач-лаборант лаборатории изучения патогенеза злокачественных опухолей 

SPIN: 2189-3404

Natalya D. Cheryarina – doctor-laboratory assistant of the laboratory for studying the pathogenesis of malignant tumors 

SPIN: 2189-3404

https://orcid.org/0000-0003-2713-8598НемашкаловаЛ. А.NemashkalovaL. A.

Немашкалова Людмила Анатольевна – научный сотрудник лаборатории изучения патогенеза злокачественных опухолей 

SPIN: 1355-8652

Lyudmila A. Nemashkalova – researcher at the laboratory for the study of the pathogenesis of malignant tumors 

SPIN: 1355-8652

https://orcid.org/0000-0001-5686-8659ЛесоваяН. С.LesovayaN. S.

Лесовая Наталья Сергеевна – младший научный сотрудник лаборатории изучения патогенеза злокачественных опухолей 

SPIN: 6995-9917

Natalya S. Lesovaya – junior researcher of the laboratory for the study of pathogenesis of malignant tumors

SPIN: 6995-9917

ФГБУ «Национальный медицинский исследовательский центр онкологии» Министерства здравоохранения Российской ФедерацииРоссияNational Medical Research Centre fоr Oncology of the Ministry of Health of RussiaRussian Federation202012032020712537Copyright © Франциянц Е.М., Каплиева И.В., Нескубина И.В., Бандовкина В.А., Трепитаки Л.К., Сурикова Е.И., Черярина Н.Д., Немашкалова Л.А., Лесовая Н.С., 20202020Франциянц Е.М., Каплиева И.В., Нескубина И.В., Бандовкина В.А., Трепитаки Л.К., Сурикова Е.И., Черярина Н.Д., Немашкалова Л.А., Лесовая Н.С.Frantsiyants E.M., Kaplieva I.V., Neskubina I.V., Bandovkina V.A., Trepitaki L.K., Surikova E.I., Cheryarina N.D., Nemashkalova L.A., Lesovaya N.S.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.rpmj.ru/rpmj/article/view/497Цель исследования

Цель исследования. Изучить особенности динамики факторов роста в условно здоровой коже, опухоли и перифокальной зоне меланомы у мышей с нокаутом по урокиназе (uPA).

Материалы и методы

Материалы и методы. Исследование проведено на разнополых мышах линий С57 ВL/6 (n=47) и C57BL/ 6-Plautm1.1BugThisPlauGFDhu/GFDhu – с нокаутом по uPA (n=31). Меланому В16/F10 перевивали под кожу в дозе 0,5 мл (1:10 в физ. растворе). Контроль – интактные мыши соответствующей линии. В коже, опухоли и перифокальной зоне, выделенных на 21‑й день роста опухоли, методом иммуноферментного анализа (ИФА) определяли уровни VEGFA, VEGFC, sVEGFR1, sVEGFR3, IGF1, IGF2, TGFβ1 и FGF21.

Результаты

Результаты. Нокаут по uPA замедлял рост (больше у самок) и метастазирование (больше у самцов) меланомы у мышей. Тормозить миграцию злокачественных клеток у самцов мог низкий уровень TGF-β1 – меньший, чем у мышей линии C57BL/6: в коже – в 5,0 раз, в опухоли – в 1,8 раза, в перифокальной зоне – в 6,1 раза. У самок с нокаутом по uPA меньшая, чем у самцов, редукция TGF-β1 в опухоли (в 1,4 раза) ограничивала метастазирование, но полностью его не подавляла – регистрировались единичные очаги в легких. Высокая концентрация IGF1 в тканях у всех мышей с нокаутом по uPA: у самцов в опухоли – в 1,4 раза, в перифокальной зоне – в 2,6 раза, в коже – в 3,6 раза, у самок в опухоли – в 2,6 раза, в перифокальной зоне – в 25,0 раз, в коже – в 13,9 раза по сравнению с мышами линии С57 ВL/6 могла поддерживать метастатический фенотип раковых клеток (у самок) или бóльшую пролиферативную активность клеток меланомы (у самцов). Низкий уровень FGF‑21 в опухоли (у самцов – в 5,3 раза, у самок – в 18,4 раза), перифокальной зоне (у самцов – в 9,6 раза, у самок – в 8,5 раза) и коже (у самцов – в 6,7 раза, у самок – в 3,3 раза) у животных с нокаутом по uPA мог быть обусловлен ростом IGF‑1, поскольку известно об их реципрокном взаимодействии. Неожиданным оказалось значительное, хотя и меньшее, чем у мышей с нормальным генотипом, накопление VEGFА в ткани меланомы: у самцов в опухоли – в 44,9 раза, в перифокальной зоне – в 6,8 раза, в коже – в 2,4 раза, у самок в опухоли – в 5,6 раза, в перифокальной зоне – в 2,6 раза, в коже – в 3,3 раза по сравнению с соответствующим интактным контролем вследствие вероятного участия рецептора uPA (uPAR) в реализации VEGF-индуцированных процессов.

Заключение

Заключение. Нокаут по гену uPA, изменяя активность системы ряда ростовых факторов, модифицирует метаболизм меланомы, замедляя ее рост и устраняя или снижая ее метастатическую активность.

Purpose of the study

Purpose of the study. Studying characteristics of the growth factor dynamics in the intact skin, tumors and perifocal tissues of melanoma in urokinase (uPA) gene-knockout mice.

Materials and methods

Materials and methods. The study included male and female С57 ВL/6 mice (n=47) and C57BL/6‑Plautm1.1BugThisPlauGFDhu/GFDhu mice with uPA gene-knockout (n=31). В16/F10 melanoma was transplanted subcutaneously at a dose of 0.5 mL (1:10 in normal saline). Intact mice of the same strain served as controls. Levels of VEGFA, VEGFC, sVEGFR1, sVEGFR3, IGF1, IGF2, TGFβ1 and FGF21 were determined by ELISA in the skin, tumor and perifocal tissues isolated on the 21st day of the tumor growth.

Results

Results. uPA gene-knockout inhibited the growth (mostly in females) and metastasis (predominantly in males) of melanoma in mice. Inhibition of the migration of malignant cells in males could be due to low levels of TGF-β1 compared to С57 ВL/6 mice: in the skin – by 5.0 times, in tumors – by 1.8 times and in perifocal tissues – by 6.1 times. In uPA gene-knockout females, lower levels of TGF-β1 were observed in tumors – by 1.4 times inhibited metastasis, but not completely, and solitary metastatic foci were registered in the lungs. Нigh levels of IGF1 in tissues of all uPA gene-knockout mice (males: in tumors by 1.4 times, in perifocal tissues by 2.6 times, in the skin by 3.6 times; females: in tumors by 2.6 times, in perifocal tissues by 25.0 times, in the skin by 13.9 times, compared to С57 ВL/6 mice) could maintain the metastatic phenotype of cancer cells (in females) or hiher proliferative activity of melanoma cells (in males). Lower levels of FGF‑21 in tumors (males – by 5.3 times, females – by 18.4 times), perifocal tissues (males – by 9.6 times, females – by 8,5 times) and skin (males – by 6.7 times, females – by 3.3 times) in uPA gene-knockout animals could be due to the IGF‑1 growth, as their reciprocal interaction is known. Interestingly, a significant, although lesser than in mice with a normal genotype, accumulation of VEGFA in melanoma tissues was observed: in males – in tumors by 44.9 times, in perifocal tissues by 6.8 times, in the skin by 2.4 times; in females – in tumors by 5.6 times, in perifocal tissues by 2.6 times, in the skin by 3.3 times, compared to the corresponding intact controls, due to the probable involvement of the uPA receptor (uPAR) in the implementation of VEGF-induced processes.

Conclusion

Conclusion. Changing the activity of a system of some growth factors, uPA gene-knockout modifies melanoma metabolism by inhibiting its growth and eliminating or reducing its metastatic activity.

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The authors declare that there are no conflicts of interest present.