ABSTRACT
In order to investigate the relationship between chromosomal radiosensitivity and early-onset cancer, the G(2) chromosomal radiosensitivity assay was undertaken on a group of 23 Danish survivors of childhood and adolescent cancer, a control group comprising their partners and a group of 38 of their offspring. In addition, the previously reported in-house control group from Westlakes Research Institute (WRI) was extended to 27 individuals. When using the 90th percentile cutoff for the WRI control group, the proportion of individuals with elevated radiosensitivity was 11, 35, 52 and 53% for the WRI control, partner control, cancer survivor and the offspring groups, respectively, with significant differences between the WRI control group and the cancer survivor group (P=0.002) and the offspring group (P<0.001). However, while the comparisons with the WRI control group support an association of chromosomal radiosensitivity with cancer predisposition, when the partner control group was used to define the radiosensitivity cutoff point, no significant differences in radiosensitivity profiles were found between the partner control group and either the cancer survivor group or the offspring group. The failure to distinguish between the G(2) aberration profiles of the apparently normal group of partners and the cancer survivor group suggests that any association with cancer should be viewed with caution, but also raises questions as to the suitability of the partners of cancer survivors to act as an appropriate control group. Heritability of the radiosensitive phenotype was examined by segregation analysis of the Danish families and suggested that 67.3% of the phenotypic variance of G(2) chromosomal radiosensitivity is attributable to a putative major gene locus with dominant effect.
Subject(s)
Chromosomes, Human/genetics , Chromosomes, Human/radiation effects , G2 Phase/radiation effects , Neoplasms/genetics , Radiation Tolerance/genetics , Adolescent , Adult , Age of Onset , Child , Child, Preschool , Chromosome Aberrations/radiation effects , Cohort Studies , DNA Damage , Female , Humans , Infant , Infant, Newborn , Male , Pilot Projects , SurvivorsABSTRACT
The G(2) chromosomal radiosensitivity assay is a technically demanding assay. To ensure that it is reproducible in our laboratory, we have examined the effects of storage and culture conditions by applying the assay to a group of healthy controls and determined the extent of intra- and inter-individual variations. Nineteen different individuals provided one or more blood samples resulting in a total of 57 successful tests. Multiple cultures from a single blood sample showed no statistically significant difference in the number of chromatid type aberrations between cultures. A 24h delay prior to culturing the lymphocytes did not significantly affect the induced G(2) score. Intra-individual variation was not statistically significant in seven out of nine individuals. Inter-individual variation was highly statistically significant (P<0.001), indicating that there is a real difference between individuals in the response to radiation using this assay.
Subject(s)
Chromosomes/radiation effects , Mutagenicity Tests/standards , Adult , Chromosome Aberrations , Female , Humans , Lymphocytes/ultrastructure , Male , Middle Aged , Reproducibility of ResultsABSTRACT
Although requiring stringent experimental conditions to achieve good reproducibility, the G2 assay has potential as a sensitive marker for cancer susceptibility, and is particularly useful in population studies. Immediate culture of blood is preferable, but overnight storage of blood either at ambient temperature or at 4 degrees C does not appear significantly to affect G2 scores. Transport of blood may lead to additional variability in assay results and should be well controlled. Although reproducibility is generally good, G2 scores on blood from certain individuals appear to show significant variability in repeat samples. Thus, determination of an individual's radiosensitivity may require multiple assays on different occasions. While it is recognized that the distinction between aligned and mis-aligned discontinuities has no scientific basis, some laboratories have decided for the purpose of record-keeping to score all aligned discontinuities as gaps, and mis-aligned discontinuities as breaks. In all cases the final G2 score should comprise the sum of all gaps and breaks.
