A Novel Berbamine Derivative Inhibits Cell Viability and Induces Apoptosis in Cancer Stem-Like Cells of Human Glioblastoma, via Up-Regulation of miRNA-4284 and JNK/AP-1 Signaling
Figure 3
BBMD3 induced apoptosis and increased cleavage of caspase-3 and PARP in GBM neurospheres.
(A) Apoptotic cells were analyzed for Annexin V-FITC reactivity and PI staining by flow cytometry. Cells from the PBT003, PBT008, PBT022 and PBT030 neurospheres were treated with BBMD3 (0, 1, 3, 5, 10 µM) for 48 hours. Apoptotic cells were identified as being reactive with Annexin V-FITC antibody (early stage of apoptosis) or PI and Annexin V-FITC/PI double-positive (late stage of apoptosis) cells. Each experiment was performed in duplicate or triplicate, and repeated twice as an independent replicate. The top of each bar graph represents the mean of the observed values from these replicates, and the error bars represent ± the standard deviation from the mean (SD). (B) The extent of cleavage of caspase-3 and PARP was determined following a 24 hour treatment with BBMD3 using Western blotting analyses. β-actin serves as an internal control.