Journal Article

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

Gomes, Luciana Rodrigues, Rocha, Clarissa Ribeiro Reily, Kinker, Gabriela Sarti, Fiore, Ana Paula Zen Petisco, Martins, Davi Jardim, Bruni-Cardoso, Alexandre, Menck, Carlos Frederico Martins

2019

Overview

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.

• Publisher: Nature Publishing Group UK,Springer Nature B.V
• Subject: 13/106 / 13/31 / 13/89 / 13/95 / 38/1 / 38/77 / 631/337/1427/2567 / 631/67/1347 / 631/67/327 / 631/80/304 / A549 Cells / Antibodies / Antineoplastic Agents - pharmacology / Antineoplastic Agents - therapeutic use / Antineoplastic drugs / Antitumor agents / Ataxia Telangiectasia Mutated Proteins - antagonists & inhibitors / Ataxia Telangiectasia Mutated Proteins - genetics / Ataxia Telangiectasia Mutated Proteins - metabolism / Autophagy / Autophagy - drug effects / Autophagy - genetics / Biochemistry / Biomedical and Life Sciences / Breast cancer / Breast Neoplasms - drug therapy / Breast Neoplasms - genetics / Breast Neoplasms - metabolism / Cell Biology / Cell Culture / Cell Culture Techniques - methods / Cell cycle / Cellular Microenvironment - drug effects / Cellular Senescence - drug effects / Checkpoint Kinase 1 - genetics / Checkpoint Kinase 1 - metabolism / Chemotherapy / CHK1 protein / Cisplatin / Cisplatin - pharmacology / Cisplatin - therapeutic use / Deoxyribonucleic acid / DNA / DNA adducts / DNA biosynthesis / DNA damage / DNA Damage - drug effects / DNA polymerase / DNA Replication - drug effects / DNA-Binding Proteins - genetics / DNA-Binding Proteins - metabolism / DNA-directed DNA polymerase / DNA-Directed DNA Polymerase - chemistry / DNA-Directed DNA Polymerase - genetics / DNA-Directed DNA Polymerase - metabolism / Drug Resistance, Neoplasm / Extracellular matrix / Female / Histones - metabolism / Humans / Immunology / Life Sciences / Lung cancer / MCF-7 Cells / p53 Protein / Phagocytosis / Phenotypes / Pyrazines - pharmacology / S Phase Cell Cycle Checkpoints - drug effects / Senescence / Sulfones - pharmacology / Tumor Cells, Cultured / Tumor Suppressor Protein p53 - genetics / Tumor Suppressor Protein p53 - metabolism