Levels of γ-H2AX Foci after Low-Dose-Rate Irradiation Reveal a DNA DSB Rejoining Defect in Cells from Human ATM Heterozygotes in Two AT Families and in Another Apparently Normal Individual

• Published in: Radiation research Vol. 166; no. 3; pp. 443 - 453
• Main Authors: Kato, Takamitsu A., Nagasawa, Hatsumi, Weil, Michael M., Little, J. B., Bedford, J. S.
• Format: Journal Article
• Language: English
• Published: United States Radiation Research Society 01-09-2006
• Subjects: Ataxia Telangiectasia - genetics; Ataxia Telangiectasia Mutated Proteins; Cell culture techniques; Cell Cycle Proteins - genetics; Cell lines; Chromosome Aberrations - radiation effects; Cultured cells; Cytogenetic Analysis - methods; DNA - genetics; DNA - radiation effects; DNA Damage; DNA Repair - genetics; DNA Repair - radiation effects; DNA-Binding Proteins - genetics; Dose-Response Relationship, Drug; Female; Fibroblasts; Fibroblasts - radiation effects; Gamma Rays; Heterozygotes; Histones - genetics; Histones - radiation effects; Humans; Hypersensitivity; Irradiation; Loss of Heterozygosity - genetics; Male; Phenotypes; Protein-Serine-Threonine Kinases - genetics; Radiation tolerance; Siblings; Space life sciences; Tumor Suppressor Proteins - genetics
• ISSN: 0033-7587; 1938-5404
• DOI: 10.1667/RR3604.1

Kato, T. A., Nagasawa, H., Weil, M. M., Little, J. B. and Bedford, J. S. Levels of γ-H2AX Foci after Low-Dose-Rate Irradiation Reveal a DNA DSB Rejoining Defect in Cells from Human ATM Heterozygotes in Two AT Families and in Another Apparently Normal Individual. Radiat. Res. 166, 443– 453 (2006). We have investigated the use of the γ-H2AX assay, reflecting the presence of DNA double-strand breaks, as a possible means for identifying individuals who are mildly hypersensitive to ionizing radiation, such as some ATM heterozygotes. We compared levels of γ-H2AX foci after irradiation in cells from six apparently normal individuals as well as from individuals from two separate AT families including the proband, mother, father and three unaffected siblings in each family. After a 1-Gy single acute (high-dose-rate) γ-ray dose delivered to noncycling contact-inhibited monolayers of cells, clear differences were seen between samples from normal individuals (ATM+/+) and probands (ATM−/−) at nearly all sampling times after irradiation, but no clear distinctions were seen for cells from normal compared to obligate heterozygotes (ATM+/−). In contrast, after 24 h of continuous irradiation at a dose rate of 10 cGy/h, appreciable differences in numbers of foci per cell were observed for cells from individuals for all the known ATM genotypes compared with controls. Four unaffected siblings had mean numbers of foci per cell similar to that for the obligate heterozygotes, whereas the other two had mean values similar to that for normal controls. We determined independently that those siblings with mean numbers of foci per cell in the range of ATM heterozygotes carried the mutant allele, while both siblings with a normal number of foci per cell after irradiation had normal alleles. A more limited set of experiments using lymphoblastoid cell strains in the low-dose-rate assay also revealed distinct differences for normal compared to ATM heterozygotes from the same families and opens the possibility of using peripheral blood lymphocytes as a more suitable material for an assay to detect mild hypersensitivities to radiation among individuals.