%0 Journal Article
%T ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation
%A Alexandre Bruni-Cardoso
%A Ana Paula Zen Petisco Fiore
%A Carlos Frederico Martins Menck
%A Clarissa Ribeiro Reily Rocha
%A Davi Jardim Martins
%A Gabriela Sarti Kinker
%A Luciana Rodrigues Gomes
%J Archive of "Cell Death & Disease".
%D 2019
%R 10.1038/s41419-019-1689-8
%X a Cellular viability analysis of cells exposure to different doses of cisplatin for 72£¿h, by XTT assays. b, c Cell death assessment in cells treated with cisplatin for 72 and 120£¿h, by fluorescence microscopy. A combination of dyes (fluorescein di-acetate, propidium iodide, and Hoechst) was used to distinguish viable cells from those in necrosis or apoptosis. c Cells were counted, and the percentages were plotted on a graph. d Cellular viability assay, in response to 72£¿h of cisplatin treatment, in MCF-7 cells cultured under 2D, 3D, or non-adherent 3D (with or without media supplemented with 4% Matrigel£¿) conditions. e Immunodetection of cisplatin¨CDNA adducts, by slot-blot assays, upon 1£¿h of treatment. f, g ¦Ã-H2AX detection, followed by flow cytometry, for evaluation of DNA damage induced by cisplatin (4£¿h). In all graphs, the results are presented as mean£¿¡À£¿SEM from at least two independent experiments performed in duplicate. Two-way analysis of variance and Bonferroni post hoc test were used for statistical analysis and the differences were considered significant for *P£¿¡Ü£¿0.05 and ***P£¿¡Ü£¿0.001. ns highlights non-significant changes. b The red arrows indicate necrotic cells (type of cell death mostly detected in MCF-7 cells) whose standard staining is characterized by the red nuclei and cytoplasm homogeneously stained with propidium iodide. The white scale bar correspond to 10£¿¦Ì
%K Breast cancer
%K Cancer microenvironment
%K Mechanisms of disease
%K DNA damage checkpoints
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6561919/