Enhancement of radiosensitivity by proteasome inhibition: Implications for a role of NF-κB

Abstract

Purpose: NF-κB is activated by tumor necrosis factor, certain chemotherapeutic agents, and ionizing radiation, leading to inhibition of apoptosis. NF-κB activation is regulated by phosphorylation of IκB inhibitor molecules that are subsequently targeted for degradation by the ubiquitin-proteasome pathway. PS-341 is a specific and selective inhibitor of the proteasome that inhibits NF-κB activation and enhances cytotoxic effects of chemotherapy in vitro and in vivo. The objective of this study was to determine if proteasome inhibition leads to enhanced radiation sensitivity. Methods and Materials: Inhibition of NF-κB activation in colorectal cancer cells was performed by treatment of LOVO cells with PS-341 or infection with an adenovirus encoding IκB super-repressor, a selective NF-κB inhibitor. Cells were irradiated at 0, 2, 4, 6, 8, and 10 Gy with or without inhibition of NF-κB. NF-κB activation was determined by electrophoretic mobility gel shift assay, and apoptosis was evaluated using the TUNEL assay. Growth and clonogenic survival data were obtained to assess effects of treatment on radiosensitization. In vitro results were tested in vivo using a LOVO xenograft model. Results: NF-κB activation was induced by radiation and inhibited by pretreatment with either PS-341 or IκBα super-repressor in all cell lines. Inhibition of radiation-induced NF-κB activation resulted in increased apoptosis and decreased cell growth and clonogenic survival. A 7–41% increase in radiosensitivity was observed for cells treated with PS-341 or IκBα. An 84% reduction in initial tumor volume was obtained in LOVO xenografts receiving radiation and PS-341. Conclusions: Inhibition of NF-κB activation increases radiation-induced apoptosis and enhances radiosensitivity in colorectal cancer cells in vitro and in vivo. Results are encouraging for the use of PS-341 as a radiosensitizing agent in the treatment of colorectal cancer.