Characterization of the epithelial and the mesenchymal breast cancer cell line BT474 and MDA-MB 436.

Abstract

<p>A) miR-200c levels in a panel of mesenchymal (MDA-MB 436, MDA-MB 231, MDA-MB 157) and epithelial (MCF-7, MDA-MB 468, BT474) breast cancer cell lines. Quantitative RT-PCR was performed to assess the levels of miR-200c. Expression was normalized to miR-191 and presented as ratio. B) Epithelial and mesenchymal phenotype of BT474 and MDA-MB 436 cells. To confirm the different phenotypes, microscopic pictures were taken from both cell lines (phase contrast, left panel). Furthermore, immunofluorescence and laser scanning microscopy was carried out to determine E-Cadherin (red, middle panel) and Vimentin (red, right panel) expression. Nuclei were stained with DAPI (blue). C) Dose reponse curves and IC₅₀ values for doxorubicin. BT474 and MDA-MB 436 cells were treated with different concentrations of doxorubicin for 72 hours. Subsequently, a CellTiter Glo assay was performed to determine the percentage of viable cells (normalized to mock treated cells). IC₅₀ values are shown in the table. Experiments were done in triplicates. For statistical analysis a student’s t-test was performed. (***p<0.001).</p