Inhibition of NF- B, Clonogenicity, and Radiosensitivity of Human Cancer Cells

Abstract

BACKGROUND: Activation of the transcription factor NF-κB is part of the immediate early response of tissues to ionizing irradiation. This pathway has been shown to protect cells from tumor necrosis factor-α, chemotherapy, and radiation therapy-induced apoptosis (programmed cell death). However, because the role of NF-κB as a modifier of the intrinsic radiosensitivity of cancer cells is less clear, we have studied the impact of NF-κB on the intrinsic radiosensitivity of human cancer cells. METHODS: We used PC3 prostate cancer cells and HD-MyZ Hodgkin's lymphoma cells transduced with an adenovirus vector that contains a gene encoding a form of IκB (an inhibitor of NF-κB) that cannot be phosphorylated. This form of IκB will remain bound to NF-κB; thus, NF-κB cannot be activated. We monitored NF-κB activity with a gel-shift assay and used a colony-forming assay to assess clonogenicity and radiosensitivity. RESULTS: Constitutive DNA-binding activity of NF-κB was dramatically decreased in PC3 cells transduced with the IκB super-repressor gene. The clonogenicity of transduced PC3 cells declined to 19.6% of that observed for untreated control cells, a finding similar to one we have previously demonstrated for IκB-transduced HD-MyZ cells. However, inhibition of NF-κB activity in the surviving PC3 and HD-MyZ cells failed to alter their intrinsic radiosensitivity. CONCLUSIONS: We conclude that activation of NF-κB does not determine the intrinsic radiosensitivity of cancer cells, at least for the cell lines tested in this study.