Detection of repair activity during the DNA damage-induced G2 delay in human cancer cells.
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.
Kao
G D
GD
Hospital of the University of Pennsylvania, Department of Radiation Oncology, 2 Donner, 3400 Spruce Street, Philadelphia, Pennsylvania, PA 19104, USA.
McKenna
W G
WG
Yen
T J
TJ
D001973:Bromodeoxyuridine; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D002503:Centromere / Q000235:genetics; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics; D004249:DNA Damage; D004260:DNA Repair; D004273:DNA, Neoplasm / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D005047:Etoposide / Q000633:toxicity; D005260:Female; D005434:Flow Cytometry; D016195:G2 Phase; D005720:Gamma Rays; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008840:Microfilament Proteins; D010051:Ovarian Neoplasms; D011836:Radiation Tolerance; D014407:Tumor Cells, Cultured
D002868:Chromosomal Proteins, Non-Histone; D004273:DNA, Neoplasm; D008840:Microfilament Proteins; C094772:centromere protein F; D005047:Etoposide; D001973:Bromodeoxyuridine