FIGURE 2.
AZD8055-mediated cytoprotection is independent of cell cycle arrest. A, DLD1 and HT-29 cells were treated with rapamycin or AZD8055 at the indicated doses for 48 h and p27Kip1 expression levels were analyzed by immunoblotting. B, DLD1 cells were transduced with a lentivirus encoding GFP or p27Kip1. Cells were then treated with 5-FU or oxaliplatin for 5 days, and expression of p-H2Ax and cleaved caspase-9 and -3 proteins were measured by immunoblotting. C, AZD8055 enhances oxaliplatin-induced autophagy, as shown by LC3I-II conversion and reduced p62/SQSTM 1 expression, which is associated with attenuated DNA damage and caspase cleavage. DLD1 cells were treated with increasing doses of oxaliplatin alone or combined with AZD8055 for 5 days and expression of LC3, p62, p-H2Ax, and cleaved caspase-9 and -3 were analyzed by immunoblotting.