PMS1077 sensitized cancer cells to TNF-α induced apoptosis and inhibited the TNF-α induced expression of anti-apoptotic gene.

<p>(A) DU145 cells were treated with vehicle, PMS1077 or PMS601 as the indicated concentration (0–80 µM) and then incubated with or without TNF-α (20 ng/ml) for 24 h. Cell viability was determined by MTT assay. (B) DU145 cells were treated with vehicle or PMS1077 (40 and 80 µM), and then incubated with TNF-α (20 ng/ml) for 24 h. Treated cells were observed by differential interference contrast microscope. (C) DU145 cells were treated as (B) and apoptosis analysis was performed by flow cytometry through annexin-V-FITC and Propidium iodide (PI) staining treated cells. (D) DU145 cells were treated with vehicle or PMS1077 (40 and 80 µM), then incubated with TNF-α (20 ng/ml) for 12 h. After treatment, the whole cell lysates were prepared and analyzed by Western blotting with antibodies as indicated. (E) DU145 cells were incubated with TNF-α (20 ng/ml) for 1 h, with vehicle or PMS1077 pretreatment for 6 h. The mRNA level of NF- κB target genes were examined using RT–PCR. (F) DU145 cells were treated as (D) and whole cell lysates were prepared and analyzed by Western blotting with antibodies for PARP and GAPDH. All data shown are representative of three independent experiments.