Figure 3
Role of caspase-9/-3 and PARP in apoptotic cell death induced by ZOL. (A) Detection of caspase-9/-3 and PARP in PC cells after ZOL treatment. Western blotting analysis was performed on protein extracts from solubilized whole cell pellets from BxPC-3 and PANC1 PC cell lines after 48 h exposure to ZOL (50 μ M) demonstrating abundant cleaved species of both caspase-9 and PARP in lysates for drug-exposed cells, while caspase-3 remained almost completely uncleaved. The experiments were performed at least three times and the results were always similar. (B) Apoptotic effects induced by ZOL in the presence of caspase inhibitors. Analysis was performed by flow cytometric immunodetection of Annexin-V staining on BxPC-3 cells after 48 h exposure to ZOL (50 μ M). VAD is a caspase-9 inhibitor, VEID is a specific caspase-6 inhibitor and DEVD is a caspase-3 inhibitor. Pretreatment with 15 μ M VAD and 15 μ M VEID significantly antagonised ZOL-induced apoptosis (P<0.005).
