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Differential Sensitivity of p53^(-) and p53⁽⁺⁾ Cells to Caffeine-induced Radiosensitization and Override of G₂ Delay¹

Departments of Radiation Oncology [S. N. P., J. S. D., P. C., M. E., W. T.] and Pathology [F. P., D. D.], Massachusetts General Hospital, Boston, Massachusetts 02114, and Massachusetts General Hospital Cancer Center, Charlestown Navy Yard, Boston, Massachusetts 02129 [S. F.]

Most drug discovery efforts have focused on finding new DNA-damaging agents to kill tumor cells preferentially. An alternative approach is to find ways to increase tumor-specific killing by modifying tumor-specific responses to that damage. In this report, we ask whether cells lacking the G₁-S arrest in response to X-rays are more sensitive to X-ray damage when treated with agents that override G₂-M arrest. Mouse embryonic fibroblasts genetically matched to be (+) or (-) p53 and rat embryonic fibroblasts (+) or (-) for wild-type p53 function were irradiated with and without caffeine, a known checkpoint inhibitor. At low doses (500 µM), caffeine caused selective radiosensitization in the p53^(-) cells. At this low dose (where no effect was seen in p53⁽⁺⁾ cells), the p53^(-) cells showed a 50% reduction in the size of the G₂-M arrest. At higher doses (2 mM caffeine), where sensitization was seen in both p53⁽⁺⁾ and p53^(-) cells, the radiosensitization and the G₂-M override were more pronounced in the p53^(-) cells. The greater caffeine-induced radiosensitization in p53^(-) cells suggests that p53, already shown to control the G₁-S checkpoint, may also influence aspects of G₂-M arrest. These data indicate an opportunity for therapeutic gain by combining DNA-damaging agents with compounds that disrupt G₂-M arrest in tumors lacking functional p53.

¹ This research was supported by NIH Grant RO1-CA58985.

² To whom requests for reprints should be addressed, at Department of Radiation Oncology, Cox 302, 100 Blossom Street, Massachusetts General Hospital, Boston, MA 02114.

³ Present address: Fred Hutchinson Cancer Research Center, Seattle, WA 98104.

Received 1/23/95. Accepted 2/23/95.

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