TNF increases camptothecin-induced apoptosis by inhibition of NF-κB

rHuTNF potentiates CPT-cytotoxicity in human ovarian A2780 cells. In this study, we examined the role of NF-kappaB in this potentiation. A pulse-labelled DNA study indicated that the combination CPT+ TNF had little effect on the rate of DNA elongation at 6 h after drug removal, whereas CPT alone produced a complete inhibition for at least 6 h after drug removal. Flow cytometry analyses showed that CPT + TNF arrested cells in the G(2)-M phase, whereas CPT blocked cells in S phase. Looking at the persistence of the NF-kappaB complexes in cells, it appeared that they were still present at 24 h in TNF-treated cells. In contrast, in CPT-treated cells they persisted for 6 h. In CPT + TNF-treated cells, the NF-kappaB complexes disappeared quickly and became undetectable at 6 h. The induction of apoptosis was detected only in the CPT+ TNF treated cells (using flow cytometry, a filter binding assay and ApopTag staining). These findings show that TNF, in combination with CPT, reduces the time that NF-kappaB complexes persist in cells likely resulting in the induction of apoptosis. (C) 2003 Elsevier Science Ltd. All rights reserved.