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ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation

Tissue architecture and cell–extracellular matrix (cell–ECM) interaction determine the organ specificity; however, the influences of these factors on anticancer drugs preclinical studies are highly neglected. For considering such aspects, three-dimensional (3D) cell culture models are relevant tools...

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Autores principales: Gomes, Luciana Rodrigues, Rocha, Clarissa Ribeiro Reily, Martins, Davi Jardim, Fiore, Ana Paula Zen Petisco, Kinker, Gabriela Sarti, Bruni-Cardoso, Alexandre, Menck, Carlos Frederico Martins
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6561919/
https://www.ncbi.nlm.nih.gov/pubmed/31189884
http://dx.doi.org/10.1038/s41419-019-1689-8
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author Gomes, Luciana Rodrigues
Rocha, Clarissa Ribeiro Reily
Martins, Davi Jardim
Fiore, Ana Paula Zen Petisco
Kinker, Gabriela Sarti
Bruni-Cardoso, Alexandre
Menck, Carlos Frederico Martins
author_facet Gomes, Luciana Rodrigues
Rocha, Clarissa Ribeiro Reily
Martins, Davi Jardim
Fiore, Ana Paula Zen Petisco
Kinker, Gabriela Sarti
Bruni-Cardoso, Alexandre
Menck, Carlos Frederico Martins
author_sort Gomes, Luciana Rodrigues
collection PubMed
description Tissue architecture and cell–extracellular matrix (cell–ECM) interaction determine the organ specificity; however, the influences of these factors on anticancer drugs preclinical studies are highly neglected. For considering such aspects, three-dimensional (3D) cell culture models are relevant tools for accurate analysis of cellular responses to chemotherapy. Here we compared the MCF-7 breast cancer cells responses to cisplatin in traditional two-dimensional (2D) and in 3D-reconstituted basement membrane (3D-rBM) cell culture models. The results showed a substantial increase of cisplatin resistance mediated by 3D microenvironment. This phenotype was independent of p53 status and autophagy activity and was also observed for other cellular models, including lung cancer cells. Such strong decrease on cellular sensitivity was not due to differences on drug-induced DNA damage, since similar levels of γ-H2AX and cisplatin–DNA adducts were detected under both conditions. However, the processing of these cisplatin-induced DNA lesions was very different in 2D and 3D cultures. Unlike cells in monolayer, cisplatin-induced DNA damage is persistent in 3D-cultured cells, which, consequently, led to high senescence induction. Moreover, only 3D-cultured cells were able to progress through S cell cycle phase, with unaffected replication fork progression, due to the upregulation of translesion (TLS) DNA polymerase expression and activation of the ATR-Chk1 pathway. Co-treatment with VE-821, a pharmacological inhibitor of ATR, blocked the 3D-mediated changes on cisplatin response, including low sensitivity and high TLS capacity. In addition, ATR inhibition also reverted induction of REV3L by cisplatin treatment. By using REV3L-deficient cells, we showed that this TLS DNA polymerase is essential for the cisplatin sensitization effect mediated by VE-821. Altogether, our results demonstrate that 3D-cell architecture-associated resistance to cisplatin is due to an efficient induction of REV3L and TLS, dependent of ATR. Thus co-treatment with ATR inhibitors might be a promising strategy for enhancement of cisplatin treatment efficiency in breast cancer patients.
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spelling pubmed-65619192019-06-21 ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation Gomes, Luciana Rodrigues Rocha, Clarissa Ribeiro Reily Martins, Davi Jardim Fiore, Ana Paula Zen Petisco Kinker, Gabriela Sarti Bruni-Cardoso, Alexandre Menck, Carlos Frederico Martins Cell Death Dis Article Tissue architecture and cell–extracellular matrix (cell–ECM) interaction determine the organ specificity; however, the influences of these factors on anticancer drugs preclinical studies are highly neglected. For considering such aspects, three-dimensional (3D) cell culture models are relevant tools for accurate analysis of cellular responses to chemotherapy. Here we compared the MCF-7 breast cancer cells responses to cisplatin in traditional two-dimensional (2D) and in 3D-reconstituted basement membrane (3D-rBM) cell culture models. The results showed a substantial increase of cisplatin resistance mediated by 3D microenvironment. This phenotype was independent of p53 status and autophagy activity and was also observed for other cellular models, including lung cancer cells. Such strong decrease on cellular sensitivity was not due to differences on drug-induced DNA damage, since similar levels of γ-H2AX and cisplatin–DNA adducts were detected under both conditions. However, the processing of these cisplatin-induced DNA lesions was very different in 2D and 3D cultures. Unlike cells in monolayer, cisplatin-induced DNA damage is persistent in 3D-cultured cells, which, consequently, led to high senescence induction. Moreover, only 3D-cultured cells were able to progress through S cell cycle phase, with unaffected replication fork progression, due to the upregulation of translesion (TLS) DNA polymerase expression and activation of the ATR-Chk1 pathway. Co-treatment with VE-821, a pharmacological inhibitor of ATR, blocked the 3D-mediated changes on cisplatin response, including low sensitivity and high TLS capacity. In addition, ATR inhibition also reverted induction of REV3L by cisplatin treatment. By using REV3L-deficient cells, we showed that this TLS DNA polymerase is essential for the cisplatin sensitization effect mediated by VE-821. Altogether, our results demonstrate that 3D-cell architecture-associated resistance to cisplatin is due to an efficient induction of REV3L and TLS, dependent of ATR. Thus co-treatment with ATR inhibitors might be a promising strategy for enhancement of cisplatin treatment efficiency in breast cancer patients. Nature Publishing Group UK 2019-06-12 /pmc/articles/PMC6561919/ /pubmed/31189884 http://dx.doi.org/10.1038/s41419-019-1689-8 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Gomes, Luciana Rodrigues
Rocha, Clarissa Ribeiro Reily
Martins, Davi Jardim
Fiore, Ana Paula Zen Petisco
Kinker, Gabriela Sarti
Bruni-Cardoso, Alexandre
Menck, Carlos Frederico Martins
ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation
title ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation
title_full ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation
title_fullStr ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation
title_full_unstemmed ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation
title_short ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation
title_sort atr mediates cisplatin resistance in 3d-cultured breast cancer cells via translesion dna synthesis modulation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6561919/
https://www.ncbi.nlm.nih.gov/pubmed/31189884
http://dx.doi.org/10.1038/s41419-019-1689-8
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