Skip to main content
Figure 6 | BMC Cancer

Figure 6

From: Enhancement of hypoxia-activated prodrug TH-302 anti-tumor activity by Chk1 inhibition

Figure 6

(A) AZD7762 enhanced apoptosis in cells co-treated with TH-302. HT29 cells were exposed to TH-302 at the indicated concentrations and 0.1 μM of AZD7762 for 2 h under either normoxia (21% O2) or hypoxia (N2). After wash, cells were continuously cultured for additional 46 h in the presence of 0.1 μM AZD7762. Caspase 3/7 activity was detected using luminescence-based caspase activity assay kit. The specificity of caspase 3/7 activity was confirmed by including pan-caspase inhibitor ZVAD. (B) AZD7762 inhibited Chk1 autophosphorylation. HT29 cells were treated with TH-302, AZD7762, or combination of TH-302 and AZD7762 for 2 h, washed, and AZD7762 added back and incubated for an additional 46 h. Cell lysates were harvested and immunoblotted with antibodies against Chk1 autophosphorylation (S296) and total Chk1. Equal loading was confirmed by actin blot. (C) TH-302 AZD7762 co-treatment abrogated cell cycle arrest biomarkers and decreased Rad51 expression. Cells were treated as described in Figure 6B, and then immunoblotted with antibodies against cell-cycle biomarkers phospho-histone H3, phospho-Cdc2 Y15, total Cdc2 and Rad51. Equal loading was confirmed by actin blot. The blot data are the representative of two independent experiments.

Back to article page