Cell and Tissue BiologyCell and Tissue BiologyЦитология0041-37713034-6061The Russian Academy of Sciences66949810.31857/S0041377124040025QDCKFTArticlesСтатьиResearch ArticleMelanoma cells adhesive properties activation in 3d spheroidsАктивация адгезивных свойств клеток меланомы в условиях 3d-культивированияChernykhD. V.ЧерныхД. В.
Russian Federation
tatyana_ruksha@mail.ruZinchenkoI. S.ЗинченкоИ. С.
Russian Federation
tatyana_ruksha@mail.ruRukshaT. G.РукшаТ. Г.
Russian Federation
tatyana_ruksha@mail.ruVoino-Yasenetsky Krasnoyarsk State Medical UniversityКрасноярский государственный медицинский университет им. проф. В.Ф. Войно-Ясенецкого, кафедра патологической физиологииru15072024202466433034027022025Received:27022025Published:15072024Copyright ©; 2024, Russian Academy of SciencesCopyright ©; 2024, Российская академия наук2024Russian Academy of SciencesРоссийская академия наукhttps://vietnamjournal.ru/0041-3771/article/view/669498

In this study, the viability and adhesive features of BRO and SK-MEL-2 cell lines were evaluated using a melanoma model. It was revealed that in BRO cells, the development of apoptosis after exposure to dacarbazine was combined with the transition of the proportion of cells to the G0 phase of the cell cycle, that corresponded to previously obtained results. The absence of apoptosis in 3D spheroids and the absence of exit from the cell cycle were observed in SK-MEL-2 melanoma cells. It was also revealed that in the control spheroids (cells without exposure) of the BRO and SK-MEL-2 melanoma lines, adhesion to fibronectin was higher compared with the cells of the control monolayer, which is explained by the three-dimensional structure requiring cell communication with the extracellular matrix. In spheroids formed by SK-MEL-2 cells, dacarbazine induced a decrease in adhesion to fibronectin, which may be associated with the development of drug resistance. An increase in the expression of integrins AV and β8 in BRO and SK-MEL-2, as well as integrin β5 in SK-MEL-2, was determined in cells after exposure to dacarbazine, which may indicate the involvement of aforementioned molecules in the exit from proliferative stage of tumor cells.

В настоящей работе на модели меланомы оценивали жизнеспособность и адгезивные свойства клеток линий BRO и SK-MEL-2. Показано, что в клетках линии BRO развитие апоптоза после воздействия дакарбазином сочеталось с переходом доли клеток в фазу G0 клеточного цикла, что подтверждает ранее полученные результаты. В клетках меланомы SK-MEL-2 наблюдали отсутствие апоптоза в 3D-сфероидах и отсутствие выхода из клеточного цикла. Также выявлено, что в контрольных сфероидах (клетки без воздействия) линий меланомы BRO и SK-MEL-2 адгезия к фибронектину была выше по сравнению с клетками монослоя контроля, что объясняется трехмерной структурой, требующей коммуникации клеток с внеклеточным матриксом. В сфероидах, сформированных клетками SK-MEL-2, дакарбазин индуцировал снижение адгезии к фибронектину, что может быть связано с развитием лекарственной устойчивости. После воздействия дакарбазином повышаются уровни экспрессии интегринов αV и β8 в клетках BRO и SK-MEL-2, а также интегрина β5 в клетках SK-MEL-2, что может указывать на участие этих молекул в утрате пролиферативного статуса опухолевых клеток.

melanomaspheroidssolid tumorsdacarbazineadhesionapoptosisfibronectinмеланомасфероидысолидные опухолидакарбазин, адгезияапоптозфибронектинРоссийский научный фондRussian Science Foundation№ 19-15-00110, https://rscf.ru/project/19-15-00110/Achill T. M., Meye J., & Morgan J. R. 2012. Advances in the formation, use and understanding of multi-cellular spheroids. Exp. Op. Biol. Ther. V. 12. P. 1347. https://doi.org/10.1517/14712598.2012.707181Aksenenko M.B., Palkina N.V., Sergeeva O.N., Sergeeva E. Yu., Kirichenko A.K., Ruksha T.G. 2019. miR-155 overexpression is followed by downregulation of its target gene, NFE2L2, and altered pattern of VEGFA expression in the liver of melanoma B16-bearing mice at the premetastatic stage. Int. J. Exp. Pathol. V. 100. P. 311. https://doi.org/10.1111/iep.12342Azimian-Zavareh V., Dehghani-Ghobadi Z., Ebrahimi M., Mirzazadeh K., Nazarenko I., Hossein G. 2021. Wnt5A modulates integrin expression in a receptor-dependent manner in ovarian cancer cells. Sci. Rep. V. 11. P. 5885. doi: 10.1038/s41598-021-85356-6Chen G., Kawazoe N., Tateishi T. 2008. Effects of ECM proteins and cationic polymers on the adhesion and proliferation of rat islet cells. The Open Biotechnol. J. V. 2: e1874070727154. https://doi.org/10.2174/1874070700802010133Colella G., Fazioli F., Gallo M., De Chiara A., Apice G., Ruosi C., Cimmino A., De Nigris F. 2018. Sarcoma spheroids and organoids — promising tools in the era of personalized medicine. Int. J. Mol. Sci. V. 19. P. 615. https://doi.org/10.3390/ijms19020615Esimbekova A.R., Palkina N.V., Zinchenko I.S., Belenyuk V.D., Savchenko A.A., Sergeeva E.Y., Ruksha T.G. 2023. Focal adhesion alterations in G0-positivemelanoma cells. Cancer Med. V. 12. P. 7294. https://doi.org/10.1002/cam4.5510Kewitz S., Stiefel M., Kramm C.M., & Staege M.S. 2014. Impact of O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation and MGMT expression on dacarbazine resistance of Hodgkin’s lymphoma cells. Leukemia research. V. 38. P. 138. https://doi.org/10.1016/j.leukres.2013.11.001Komina A.V., Palkina N.V., Aksenenko M.B., Lavrentev S.N., Moshev A.V., Savchenko A.A., Averchuk A.S., Rybnikov Y.A., Ruksha T.G.2019.Semaphorin-5A downregulation is associated with enhanced migration and invasion of BRAF-positive melanoma cells under vemurafenib treatment in melanomas with heterogeneous BRAF status. Melanoma Res. V. 29. P. 544. https://doi.org/10.1097/CMR.0000000000000621Lee S. Y., Koo I. S., Hwang H. J., Lee D. W. 2023. In vitro three-dimensional (3D) cell culture tools for spheroid and organoid models. SLAS Discov. V. 29. P. 5 131. https://doi.org/10.1016/j.slasd.2023.03.006Lin T. C., Yang C. H., Cheng L. H., Chang W. T., Lin Y. R., Cheng H.C. 2019. Fibronectin in cancer: friend or foe. Cells. V. 9. P. 27. doi: 10.3390/cells9010027.Livak K. J., Schmittgen T. D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. V. 25. P. 402. https://doi.org/10.1006/meth.2001.1262Liu Y., Sheikh M. S. 2014. Melanoma: molecular pathogenesis and therapeutic management. Mol. Cell.Pharmacol. V. 6. P. 228.Sakalem M. E., De Sibio M. T., da Costa F. A. D. S., de Oliveira M. 2021. Historical evolution of spheroids and organoids, and possibilities of use in life sciences and medicine. Biotechnol. J. V. 16: 2000463. https://doi.org/10.1002/biot.202000463Srisongkram T., Weerapreeyakul N., Thumanu K. 2020. Evaluation of melanoma (SK-MEL-2) cell growth between three-dimensional (3D) and two-dimensional (2D) cell cultures with fourier transform infrared (FTIR) microspectroscopy. Int. J. Mol. Sci. V. 21: 4141. https://doi.org/10.3390/ijms21114141Syed A. M., Kundu S., Ram C., Kulhari U., Kumar A., Mugale M. N., Murty U.S., Sahu B. D. 2022. Aloin alleviates pathological cardiac hypertrophy via modulation of the oxidative and fibrotic response. Life Sci., V. 288: 120159. https://doi.org/10.1016/j.lfs.2021.120159Zanoni M., Cortesi M., Zamagni A., Arienti C., Pignatta S., Tesei A. 2020. Modeling neoplastic disease with spheroids and organoids. J. Hematol. Oncol. V. 13. P. 1. https://doi.org/10.1186/s13045-020-00931-0Zanoni M., Piccinini F., Arienti C., Zamagni A., Santi S., Polico R., Bevilacqua A., Tesei A. 2016. 3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained. Sci. Rep. V. 6: 19103. https://doi.org/10.1038/srep19103Zhou J., Yi Q., Tang L. 2019. The roles of nuclear focal adhesion kinase (FAK) on cancer: a focused review. J. Exp. Clin. Cancer Res. V. 38. P. 250. https://doi: 10.1186/s13046-019-1265-1Wu C., Weis S. M., Cheresh D. A. 2023. Upregulation of fibronectin and its integrin receptors – an adaptation to isolation stress that facilitates tumor initiation. J. Cell Sci. V. 136: 261483. https://doi.org/10.1242/jcs.261483