Figure 4
In vitro and in vivo effect of AS602868 combined with CPT-11/SN-38 on apoptosis. (A) Apoptosis was measured by ELISA. HT-29 cells were incubated for 5 days with AS602868±SN-38 at indicated concentrations. Data are expressed as mean OD±s.d. of duplicates of one representative experiment from 4. The positive control is a DNA–histone complex included in the kit. (B) Tumours were removed from mice treated with AS602868 vehicle (control group), with AS602868 (20 mg kg−1) alone or combined with CPT-11 (30 mg kg−1). Then, they were minced, put into liquid nitrogen, and stored at −80°C. Frozen tumour sections (7 μm) were mounted in Fluoromount-G solution and processed following the protocol described in the In situ Cell Death Detection Kit (Roche Diagnostics). Analyses were performed using an LSM 510 confocal laser-scanning microscope with an oil objective × 40. (C) HT-29 cells were treated with AS602868±SN-38±z-VADfmk (50 μ M) for 5 days. Cell viability was evaluated using the MTT assay. Data are expressed as means±s.d. of quadruplicates of one representative experiment from 3.
