Figure 5.

Role of NF-κB in tumor cell sensitivity to gemcitabine alone or in combination with Ad-dCK::UMK and TS/RRM2 siRNA. BxPc3 and Panc1 tumor cells were cotreated with Ad-dCK::UMK and TS/RR siRNA. One day later, cells further received 2 and 40 µM of gemcitabine, respectively, and NF-κB activation was studied after 48 hours. (A) NoShift NF-κB colorimetric assay. Cells were harvested, and NF-κB activity was evaluated according to the manufacturer's recommendations. (B) NF-κB luciferase gene reporter assay. Herein, cells were primary transfected with a plasmid-expressing luciferase under control of an NF-κB response element. Luciferase assays were then performed according to the manufacturer's protocol. All experiments were performed in triplicate and repeated at least three times. Asterisks indicate significant difference (**P < .01 and ***P < .001) observed in cells treated with the tritherapy gemcitabine plus Ad-dCK::UMK plus TS/RR siRNA compared with cells treated with gemcitabine alone. *P < .05 indicates the significant difference observed between gemcitabine-treated Panc1 cells relative to untreated Panc1 cells.