Activation of AMP-Activated Protein Kinase and Extracelluar Signal-Regulated Kinase Mediates CB-PIC-Induced Apoptosis in Hypoxic SW620 Colorectal Cancer Cells
Cells were treated with or without CB-PIC (0 or 40 g/mL) under normoxic or hypoxic conditions for 6 hr. (a) Cell lysates were prepared and subjected to Western blotting to determine the expression of AMPKα, pAMPK, PARP, BCl-2, pERK, ERK, and -actin. Band density of pAMPK, cleaved PARP, BCl-2, and pERK were quantified using Gel-pro analyzer (Media Cybernetics, Bethesda, MD, USA). Values are means SD, . and compared with normoxia and hypoxia groups. (b) Cells ( cells) treated with CB-PIC for 6 hours were measured enzyme activity of the caspase-3 class of protease in apoptotic cells by using Caspase-3 Colorimetric Assay Kit. (c) Cells were transiently transfected with AMPK siRNA or control siRNA in the presence or absence of CB-PIC (40 g/mL), and PD 98059 was also treated to SW 620 in the presence or absence of CB-PIC (40 g/mL) for 6 hours in hypoxia. Western blotting was performed to determine the expression of PARP, AMPK, pAMPK, pERK, ERK, and -actin. (d) TUNEL staining was performed and visualized under fluorescence microscopy (200).