Deguelin Induces Both Apoptosis and Autophagy in Cultured Head and Neck Squamous Cell Carcinoma Cells
Figure 5
Deguelin activates AMPK signaling.
Hep-2 cells were exposed to 100 µM of deguelin for indicated time points, followed by western blot detecting phospho- and total- level of LKB1-AMPK-ACC using specific antibodies (A). Hep-2 cells were transfected with scramble or AMPKα1RNAi (100 nM each) for 48 hours. Western blot was utilized to test AMPKα1 expression after transfection. Successfully AMPK knockdown cells and their parental cells were treated with deguelin (100 µM) in the presence or absence of AICAR (1 mM), phospho- and total- level of AMPKα1 were analyzed in (B–C), MTT assay was then used to test cell viability after 48 hours (D). Hep-2 cells were pretreated with fumonisin B1 (20 µM), C6-ceramide (10 µg/ml) or NAC (500 µM) for 1 hr before deguelin exposure for indicated time points, phospho- and total level of AMPKα1 as well as tubulin were tested by western blots (E–F). The mean of at least three independent experiments performed in triplicate is shown. Statistical significance was analyzed by ANOVA.