Journal: Nature Communications
Article Title: Simultaneous CK2/TNIK/DYRK1 inhibition by 108600 suppresses triple negative breast cancer stem cells and chemotherapy-resistant disease
doi: 10.1038/s41467-021-24878-z
Figure Lengend Snippet: A Schematic diagram showing the strategy used for the generation of paclitaxel-resistant TNBC cell lines. MDA-MB-231 or BT-20 cells were grown in increasing concentrations of paclitaxel and GI 50 values were determined after every passage. Each subsequent passage was maintained in the highest possible concentration of paclitaxel-containing medium. Selection was completed when resistance reached > 200-fold; B GI 50 values for paclitaxel and 108600 for parental, paclitaxel-sensitive (TS) and paclitaxel-resistant (TR) MDA-MB-231 and BT-20 cells using CellTiter Blue in conjunction with clonogenic assays. C Expression of MDR-1, CD44, CK2, DYRK1A, EGFR, β-Catenin in paclitaxel-sensitive and resistant MDA-MB-231 cells as determined by western blot analysis. GAPDH serves as a loading control. Images are representative of two independent experiments. Samples derive from the same experiment and the blots processed in parallel. D Paclitaxel-resistant MDA-MB-231 cells were treated with increasing concentrations of 108600 for 24 h. Lysates derived from these cells were subjected to western blot analysis using the indicated antibodies. The blot is representative of three independent experiments. Samples derive from the same experiment and the blots processed in parallel. E Paclitaxel-resistant MDA-MB-231 cells were treated with increasing concentrations of paclitaxel or 108600 for 72 h, washed and allowed to grow for 7 days in drug-free growth medium. Colonies were stained with crystal violet. Source data are provided as a source data file.
Article Snippet: TNBC cell lines, sorted BCSCs from cell lines and patient-derived human breast cancer stem cells (XLC479, Creative Bioarray) were cultured in suspension in serum free DMEM/F12 1:1 supplemented with EGF (20 ng/ml, BD Biosciences), B27 (1:50, Invitrogen), 0.4% bovine serum albumin (Sigma–Aldrich), and 4 μg/ml insulin (Sigma–Aldrich) on Costar ultralow attachment multi-well plates (Corning Cat#3471).
Techniques: Concentration Assay, Selection, Expressing, Western Blot, Control, Derivative Assay, Staining
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