Fig. 8.

The effect of PtPz4, PtPz6, cisPt, and anti-MUC1 on caspase-9 mRNA in DLD-1 and HT-29 cells (A), pro- and active caspase-9 in DLD-1 (B), pro- and active caspase-9 in HT-29 cells (C) caspase-3 mRNA in DLD-1 and HT-29 cells (D), and pro-caspase-3 protein in both colon cancer cells (E). The cells were incubated for 24 h with PtPz4 (10 μM), PtPz6 (10 μM), cisPt (10 μM), anti-MUC1 (5 μg/mL), PtPz4 + anti-MUC1 (10 μM + 5 μg/mL), PtPz6 + anti-MUC1 (10 μM + 5 μg/mL), and cisPt + anti-MUC1 (10 μM + 5 μg/mL). mRNAs were determined by RT-PCR. The result is presented as a relative fold change in mRNA expression of gene in comparison of gene in control where expression was set as 1. ±SD are the mean of triplicate cultures. *P < 0.05, **P < 0.01, ***P < 0.001 compared to untreated control; ^##P < 0.01, ^###P < 0.001 compared to PtPz4 monotherapy; ^††P < 0.01 compared to cisPt monotherapy; ^‡P < 0.05, ^‡‡P < 0.01, ^‡‡‡P < 0.001 compared to anti-MUC1 monotherapy. Pro- and active caspase-9 and pro-caspase-3 expressions in cell lysates of both cell lines were evaluated by Western blotting. The intensities of the bands were quantified by densitometry. Data show the mean ±SD of the relative expression levels (from 3 assays) standardized to β-actin. *P < 0.05, **P < 0.01, ***P < 0.001 compared to the untreated control; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 compared to PtPz4 monotherapy; ^ΔΔP < 0.01, ^ΔΔΔP < 0.001 compared to PtPz6 monotherapy; ^††P < 0.01, ^†††P < 0.001 compared to cisPt monotherapy; ^‡P < 0.05, ^‡‡P < 0.01 compared to anti-MUC1 monotherapy.