Susceptibility to Radiation Induced Apoptosis and Senescence in p53 Wild Type and p53 Mutant Breast Tumor Cells
Summary rept. 1 Jul 2003-30 Jun 2006
VIRGINIA COMMONWEALTH UNIV RICHMOND
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1,25-dihydroxyvitamin D3 1,25OH2D3 and vitamin D3 analogs such as EB 1089 potentiate the response to ionizing radiation in breast tumor cells. The current studies address the basis for this interaction by evaluating DNA damage and repair, the impact of interference with reactive oxygen generation, the involvement of p53 and caspase 3, signaling through c-myc, as well as the induction of senescence and multiple modes of cell death. EB 1089 failed to increase the extent of radiation-induced DNA damage or to attenuate the rate of DNA repair. The reactive oxygen scavengers N-acetyl cysteine and reduced glutathione failed to protect the cells from the promotion of cell death by EB 1089 and radiation. While MCF-7 cells expressing caspase 3 demonstrated significant apoptosis with radiation alone as well as with EB 1089 followed by radiation, EB 1089 maintained its ability to confer susceptibility to radiation-induced cell killing, in large part by interference with proliferative recovery. In contrast, in breast tumor cells lacking p53, where radiation promoted extensive apoptosis and the cells failed to recover after radiation treatment, EB 1089 failed to influence the impact of radiation. EB 1089 treatment interfered with radiation-induced suppression of c-myc however, induction of c-myc did not prevent senescence by radiation alone or radiation-induced cell death promoted by EB 1089. EB 1089 did not increase the extent of micronucleation, indicative of mitotic catastrophe, induced by radiation alone. However, EB 1089 did promote extensive autophagic cell death in the irradiated cells. Taken together, these studies suggest that the impact of EB 1089 treatment on the radiation response is related, in part, to enhanced promotion of autophagic cell death and in part to interference with the proliferative recovery that occurs with radiation alone in p53 wild-type breast tumor cells.
- Medicine and Medical Research