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Down-Regulation of Nuclear Factor B Is Required for p53-dependent Apoptosis in X-Ray-irradiated Mouse Lymphoma Cells and Thymocytes¹

Department of Regulatory Radiobiology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553 [H. K., Y. Y., M. T., F. S.]; Radiation Biology Center, Kyoto University, Kyoto 606-8501 [O. N.]; and Cancer Research Institute, Kanazawa University, Kanazawa 920-0934 [K-i. Y.], Japan

Transcription factors p53 and nuclear factor B (NF-B) have been implicated in apoptosis induced by DNA-damaging agents, but the relationship between these two factors at the molecular level is largely unknown. We have isolated apoptosis-resistant mutant sublines from a radiosensitive mouse lymphoma 3SB cell line that undergoes p53-depen-dent apoptosis after X-ray irradiation, and we have analyzed the NF-B activity. Two of these apoptosis-resistant sublines expressed mutant p53 protein and exhibited a defect in the induction of cyclin-dependent kinase inhibitor p21 after X-ray irradiation. A decrease in the DNA binding activity of NF-B was observed in the parental 3SB cells after exposure to X-rays, whereas the same activity was unaffected by radiation in the two mutant sublines. A similar down-regulation of NF-B activity by X-rays was observed in thymocytes derived from p53 wild-type and heterozygous mice, but not in thymocytes from p53 homozygous knockout mice. These results suggest that NF-B inactivation is p53 dependent and is required for X-ray-induced apoptosis in thymic lymphoma cells and normal thymocytes.

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