Fig. 8

SS tumor growth inhibition and apoptotic cell death are enhanced by co-treatment with SAHA and SST0001. a Growth curves of CME-1 xenografts grown in the leg muscle of mice. Animals were treated with vehicle (controls), or SST0001 s.c. at 60 mg/kg, 2qdx5/w for 4 weeks, or SAHA by oral gavage at 100 mg/kg, qdx5/w for 4 weeks, or with the two drugs in combination, starting 1 day after tumor cell injection. Each point is the mean tumor volume in 6/8 mice ± SD. *P < 0.05 referred to the entire curves and the last time point. b, d CME-1 cells were treated with SAHA (0.8 and 1.6 μM) and SST0001 (0.5 mg/ml) alone or in combination at the indicated times. The effect of treatments on ERK and AKT activation, EGR1 expression, caspase 3 and PARP cleavage (b) and on heparanase expression (d) was analyzed by Western blotting. Vinculin and actin are shown as loading controls in immunoblots. Numbers represent the intensity of relevant bands normalized with respect to the respective loading controls. c Cells were exposed to SAHA (1.6 μM) and SST0001 (0.5 mg/ml), alone or in combination for 72 h, to detect caspase 3 cleavage by Western blotting and apoptosis by cytoplasmic histone-associated DNA fragmentation assay. Bars represent mean values referred to control cells ± SE obtained in four independent biological replicates. *P < 0.05 vs single agents and controls