Abstract
All eukaryotic cells manifest cell cycle delay after exposure to DNA damaging agents. It has been proposed that such cell cycle checkpoints may allow DNA repair but direct evidence of such activity during the radiation-induced G2 delay has been lacking. We report here that cells arrested in G2 by radiation (2–3 Gy) and etoposide incorporate bromodeoxyuridine (BrdU) at discrete foci in the nucleus. We detected G2 cells with CENP-F, a nuclear protein maximally expressed in G2. Caffeine and okadaic acid, both established radiosensitizers, inhibit the incorporation of BrdU in G2 cells. Radioresistant HT29 and OVCAR cells demonstrate BrdU foci formation more frequently during the G2 delay when compared to the more radiosensitive A2780 cell line. The repair foci formed during G2 may be followed through mitosis and observed in daughter cells in G1. Taken together, these observations are consistent with the detection of DNA repair activity during the radiation-induced G2 delay after relatively low doses of radiation.
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Acknowledgements
We would like to thank members of the Yen lab for invaluable discussions and comments on the manuscript, and to D Chapman and C Stobbe for assistance with the irradiation. Ms Roseanne Diehl superbly assisted the preparation of this manuscript. TJ Yen was supported by grants from the NIH, PO1 core grant CA06927, and an Appropriation from the Commonwealth of Pennsylvania.
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Kao, G., McKenna, W. & Yen, T. Detection of repair activity during the DNA damage-induced G2 delay in human cancer cells. Oncogene 20, 3486–3496 (2001). https://doi.org/10.1038/sj.onc.1204445
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DOI: https://doi.org/10.1038/sj.onc.1204445
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