THE ROLE OF EPITHELIAL-TO-MESENCHYMAL TRANSITION AND AUTOPHAGY IN ANTITUMORAL RESPONSE OF MELANOMA CELL LINES TO TARGET INHIBITION OF MEK AND mTOR KINASES

Introduction. Cutaneous melanoma is a challenge to treat due to rapid progression of disease and acquired resistance to therapy. Autophagy and the epithelial-to-mesenchymal transition (EMT) are closely interrelated and play a key role in tumor progression. Targeted co-inhibition of MEK and mTOR kina...

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Main Authors: A. S. Lozhkomoev, O. V. Bakina, A. N. Fomenko, A. V. Avgustinovich, S. G. Afanasyev, A. Yu. Dobrodeev, L. V. Spirina, A. S. Tarasova, U. B. Urmonov
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Language:Russian
Published: Russian Academy of Sciences, Tomsk National Research Medical Center 2019-06-01
Series:Сибирский онкологический журнал
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Online Access:https://www.siboncoj.ru/jour/article/view/1097
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author A. S. Lozhkomoev
O. V. Bakina
A. N. Fomenko
A. V. Avgustinovich
S. G. Afanasyev
A. Yu. Dobrodeev
L. V. Spirina
A. S. Tarasova
U. B. Urmonov
author_facet A. S. Lozhkomoev
O. V. Bakina
A. N. Fomenko
A. V. Avgustinovich
S. G. Afanasyev
A. Yu. Dobrodeev
L. V. Spirina
A. S. Tarasova
U. B. Urmonov
author_sort A. S. Lozhkomoev
collection DOAJ
description Introduction. Cutaneous melanoma is a challenge to treat due to rapid progression of disease and acquired resistance to therapy. Autophagy and the epithelial-to-mesenchymal transition (EMT) are closely interrelated and play a key role in tumor progression. Targeted co-inhibition of MEK and mTOR kinases is a potential target for melanoma therapy by downregulatoin of the EMT.Objective: to study the effect of MEK and mTOR co-inhibition on cell viability, ability to form 3D-spheroids and migratory capacity of melanoma cell lines, and correlation of these changes with EMTand autophagy-related markers.Material and Methods. Melanoma cell lines Mel Z and Mel MTP were derived from patients, who were treated at the N.N. Blokhin National Medical Research Center of Oncology. The antiproliferative effect of binimetinib and/or rapamycin was studied by the MTT -test. 3D spheroids were formed using RGD peptides. Cell migration and invasion were assessed by a Boyden chamber migration assay. The expression levels of autophagy and EMT markers were investigated by immunocytochemistry or immunoblotting.Results. Rapamycin increased cytotoxicity of binimetinib in both 2D and 3D melanoma cell line cultures. At the same time, binimetinib and rapamycin reduced invasion, but not migration capacity of melanoma cells in vitro. The effectiveness of the combination was associated with a decrease in the EMT markers (N-cadherin and β-catenin) and autophagy markers (Beclin 1, p62/SQST M1 and LC3BII ) in melanoma cells.Conclusion. Inactivation of autophagy and EMT leads to overcoming the resistance to current anti-melanoma therapy and can be considered as a promising target for the treatment of melanoma.
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spelling doaj-art-97d52665bdd14001be9cffa09cdba62a2025-08-20T03:56:24ZrusRussian Academy of Sciences, Tomsk National Research Medical CenterСибирский онкологический журнал1814-48612312-31682019-06-01183647010.21294/1814-4861-2019-18-3-64-70643THE ROLE OF EPITHELIAL-TO-MESENCHYMAL TRANSITION AND AUTOPHAGY IN ANTITUMORAL RESPONSE OF MELANOMA CELL LINES TO TARGET INHIBITION OF MEK AND mTOR KINASESA. S. Lozhkomoev0O. V. Bakina1A. N. Fomenko2A. V. Avgustinovich3S. G. Afanasyev4A. Yu. Dobrodeev5L. V. Spirina6A. S. Tarasova7U. B. Urmonov8Institute of Strength Physics and Materials Science of the Siberian Branch of the Russian Academy of SciencesInstitute of Strength Physics and Materials Science of the Siberian Branch of the Russian Academy of SciencesInstitute of Strength Physics and Materials Science of the Siberian Branch of the Russian Academy of SciencesTomsk National Research Medical Center, Russian Academy of SciencesTomsk National Research Medical Center, Russian Academy of SciencesTomsk National Research Medical Center, Russian Academy of SciencesTomsk National Research Medical Center, Russian Academy of SciencesTomsk National Research Medical Center, Russian Academy of SciencesTomsk National Research Medical Center, Russian Academy of SciencesIntroduction. Cutaneous melanoma is a challenge to treat due to rapid progression of disease and acquired resistance to therapy. Autophagy and the epithelial-to-mesenchymal transition (EMT) are closely interrelated and play a key role in tumor progression. Targeted co-inhibition of MEK and mTOR kinases is a potential target for melanoma therapy by downregulatoin of the EMT.Objective: to study the effect of MEK and mTOR co-inhibition on cell viability, ability to form 3D-spheroids and migratory capacity of melanoma cell lines, and correlation of these changes with EMTand autophagy-related markers.Material and Methods. Melanoma cell lines Mel Z and Mel MTP were derived from patients, who were treated at the N.N. Blokhin National Medical Research Center of Oncology. The antiproliferative effect of binimetinib and/or rapamycin was studied by the MTT -test. 3D spheroids were formed using RGD peptides. Cell migration and invasion were assessed by a Boyden chamber migration assay. The expression levels of autophagy and EMT markers were investigated by immunocytochemistry or immunoblotting.Results. Rapamycin increased cytotoxicity of binimetinib in both 2D and 3D melanoma cell line cultures. At the same time, binimetinib and rapamycin reduced invasion, but not migration capacity of melanoma cells in vitro. The effectiveness of the combination was associated with a decrease in the EMT markers (N-cadherin and β-catenin) and autophagy markers (Beclin 1, p62/SQST M1 and LC3BII ) in melanoma cells.Conclusion. Inactivation of autophagy and EMT leads to overcoming the resistance to current anti-melanoma therapy and can be considered as a promising target for the treatment of melanoma.https://www.siboncoj.ru/jour/article/view/1097melanoma. autophagyepithelial-to-mesenchymal transition3d spheroidsexperimental therapy
spellingShingle A. S. Lozhkomoev
O. V. Bakina
A. N. Fomenko
A. V. Avgustinovich
S. G. Afanasyev
A. Yu. Dobrodeev
L. V. Spirina
A. S. Tarasova
U. B. Urmonov
THE ROLE OF EPITHELIAL-TO-MESENCHYMAL TRANSITION AND AUTOPHAGY IN ANTITUMORAL RESPONSE OF MELANOMA CELL LINES TO TARGET INHIBITION OF MEK AND mTOR KINASES
Сибирский онкологический журнал
melanoma. autophagy
epithelial-to-mesenchymal transition
3d spheroids
experimental therapy
title THE ROLE OF EPITHELIAL-TO-MESENCHYMAL TRANSITION AND AUTOPHAGY IN ANTITUMORAL RESPONSE OF MELANOMA CELL LINES TO TARGET INHIBITION OF MEK AND mTOR KINASES
title_full THE ROLE OF EPITHELIAL-TO-MESENCHYMAL TRANSITION AND AUTOPHAGY IN ANTITUMORAL RESPONSE OF MELANOMA CELL LINES TO TARGET INHIBITION OF MEK AND mTOR KINASES
title_fullStr THE ROLE OF EPITHELIAL-TO-MESENCHYMAL TRANSITION AND AUTOPHAGY IN ANTITUMORAL RESPONSE OF MELANOMA CELL LINES TO TARGET INHIBITION OF MEK AND mTOR KINASES
title_full_unstemmed THE ROLE OF EPITHELIAL-TO-MESENCHYMAL TRANSITION AND AUTOPHAGY IN ANTITUMORAL RESPONSE OF MELANOMA CELL LINES TO TARGET INHIBITION OF MEK AND mTOR KINASES
title_short THE ROLE OF EPITHELIAL-TO-MESENCHYMAL TRANSITION AND AUTOPHAGY IN ANTITUMORAL RESPONSE OF MELANOMA CELL LINES TO TARGET INHIBITION OF MEK AND mTOR KINASES
title_sort role of epithelial to mesenchymal transition and autophagy in antitumoral response of melanoma cell lines to target inhibition of mek and mtor kinases
topic melanoma. autophagy
epithelial-to-mesenchymal transition
3d spheroids
experimental therapy
url https://www.siboncoj.ru/jour/article/view/1097
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