Chelidonine Induces Caspase-Dependent and Caspase-Independent Cell Death through G2/M Arrest in the T98G Human Glioblastoma Cell Line
Figure 4
Chelidonine-mediated arrest induces apoptosis in T98G cells. (a) Chelidonine-mediated multipolar spindle assembly formation. T98G cells were synchronized at G1/0 through double thymidine inhibition. After the synchronization, the cells were released and cultured in the presence or absence of 0.6 μM chelidonine for 12 or 18 h. Cells were immunostained for α-pericentrin (green), α-tubulin (red), and DNA (DAPI; blue). Images were captured using confocal laser scanning microscope. Magnification, 600×. (b) Whole-cell T98G lysates were subjected to western blot analysis with antibodies against cyclin B1, total or phosphorylated CDK1 (Tyr15 and Thr161), aurora A, total and phosphorylated PLK-1 (Thr210). Tubulin served as a loading control. (c) After the synchronization, cells were cultured in the presence or absence of 10 μM RO-3306 and/or 0.6 μM chelidonine for 24 h. And then the whole-cell T98G lysates were subjected to western blot analysis with the indicated antibodies. (d and e) After the synchronization, cells were cultured in the presence or absence of 10 μM RO-3306 and/or 0.6 μM chelidonine for 24 h. And then the mitochondrial potential (d) and the size of sub G1/0 population (e) of T98G cells were determined by flow cytometry analysis. Each experimental result is shown as the mean and SEM of three independent experiments. The data was analyzed using t-test. p < 0.05, p < 0.01.