Figure 5

NF-κB Inhibition Differentially Sensitizes Thyroid Cancer Cell Lines to TNFα-induced Apoptosis. (A) BCPAP, SW1736, TPC1, and C643 cells were transduced with either Ad-GFP or Ad-mIκBα at an MOI of 200, 100, 50, and 100, respectively. After 24 hours, cells were treated with TNFα (10 ng/ml) or vehicle for 3 days. Cell viability was then assessed by MTS assay. Data is reported as relative cell viability (%) compared to vehicle-treated cells (normalized to 100%), which is denoted by the dashed line. Mean ± S.E.M. of 3 independent experiments performed in octuplicate is reported. [p < 0.05 (*); p < 0.01 (**); p < 0.001 (***)] (B) BCPAP, SW1736, TPC1, and C643 cells were transduced with either Ad-GFP or Ad-mIκBα at an MOI of 200, 100, 50, and 100, respectively. After 24 hours, cells were treated with TNFα (10 ng/ml) for 15 min. Nuclear fractions were prepared, and nuclear levels of p65 were analyzed by Western blot analysis. PARP was used as a nuclear loading control. (C) SW1736 and TPC1 cells were transduced with either Ad-GFP or Ad-mIκBα at an MOI of 100 and 50, respectively. After 24 hours, the cells were serum-starved overnight and then treated with 10 ng/ml TNFα for 0, 4, 8, and 16 hours. Whole-cell extracts were harvested, and Western blot analysis was performed to assess levels of cleaved PARP (denoted as PARP) as a marker of apoptosis.