Cabozantinib induces cell-cycle arrest, polyploidy, and cell death
(A) 33, 33R, 33xR, and 33RR cells were treated with increasing concentrations of cabozantinib for 48 h. Viable cells were evaluated with ADAM assays. N = 4.
(B) 33, 33R, 33xR, and 33RR cells were treated with cabozantinib 2.5 or 5 μM, and clonogenic potential was evaluated after 10 days. Representative images are shown (see Figure S3D). N = 4.
(C) 33, 33R, 33xR, and 33RR cells were treated with cabozantinib 5 μM for 24 h. The cell cycle was analyzed by cytometry. N = 3.
(D–F) 33, 33R, 33xR, and 33RR cells were treated with cabozantinib 5 μM for 48 h.
(D) Cells were labeled for 15 min with propidium iodide (PI) and analyzed by flow cytometry. Histograms represent the percentage of cells with a DNA content of 4, 8, and 16N. N = 3.
(E) Plk1, PARP, and cleaved PARP (reflecting apoptosis) were detected by immunoblotting. HSP90 served as a loading control. N = 3.
(F) Cells were stained with PI/AnnexinV (AV). Histograms show AV+/PI- cells (apoptosis) and AV+/PI+ cells (post-apoptosis and/or another cell death). N = 3. Statistics were performed using ANOVA test: ∗∗p < 0.01, ∗∗∗p < 0.001.