Increased NF-κB DNA binding but not transcriptional activity during apoptosis induced by the COX-2-selective inhibitor NS-398 in colorectal carcinoma cells

Nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit colorectal neoplasia, an effect that is associated with their ability to induce apoptosis. Although NSAIDs have been reported to inhibit NF-κB, more recent studies show activation of NF-κB by NSAIDs. NF-κB commonly shows antiapoptotic activity and is implicated in the therapeutic resistance of cancer cells. The effects of highly COX-2-selective NSAIDs such as NS-398 on NF-κB in colorectal tumour cells have not been reported. Therefore, we addressed whether NF-κB has a role in NS-398-induced apoptosis of colorectal cancer cells. Treatment of HT-29 colorectal carcinoma cells with doses of NS-398 (50–75 μM) known to induce apoptosis had no effect on NF-κB for up to 48 h. However after 72 and 96 h NF-κB DNA-binding activity was increased by NS-398, in parallel with apoptosis induction. NS-398-treated HT-29 cells showed increased p50 homodimer binding and an induction of p50/p65 heterodimers, as demonstrated by supershift assay. However, although NS-398 increased NF-κB DNA binding it did not increase NF-κB-dependent reporter activity and inhibition of NF-κB DNA binding did not enhance NS-398-induced apoptosis. This indicates that NF-κB activated by NS-398 is transcriptionally inactive and is an encouraging result for the use of COX-2-selective NSAIDs not only in chemoprevention but also as novel therapies for colon cancer.