The chromosome-based challenge assay using fluorescence in situ hybridization: a possible test for increased cancer susceptibility

Abstract

The challenge assay is a cytogenetic approach to measure the repair competence of cells. For in vitro studies, human lymphocytes are exposed to different substances and are irradiated simultaneously. To investigate subjects exposed occupationally or environmentally, untreated blood samples are directly irradiated without any further treatment. Certain substances like heavy metals reveal carcinogenic potential without well defined mechanism of action. While they are not mutagenic they may have an effect on DNA repair capacity. The challenge assay was successfully applied in vitro experiments with cadmium to detect an interaction of this heavy metal with the repair of X-ray-induced chromosome breaks. CdCl 2 alone had no effect on the formation of chromosome aberrations (CA), not even in the cytotoxic concentration (50 μM). However, cadmium showed an effect on the number of chromosomal rearrangements (CR) after X-ray challenge. For 0.5 μM CdCl 2, CA frequencies were significantly elevated compared to the rates for X-rays alone. For the two higher concentrations the rates showed a slight additional increase. Hence, the challenge assay appears suitable to test for chromosomal sensitivity induced by toxicants. Subsequently, a study of styrene exposed workers was initiated to address the question whether styrene exposure has an influence on the DNA repair. In addition, we investigated whether a polymorphism of genes coding for phase II detoxifying enzymes glutathione– S-transferases GSTM1 and GSTT1 had an influence on chromosomal sensitivity. First and preliminary data are presented. While there is a correlation of the rate of CR with cumulative lifetime exposure of styrene, the most recent styrene exposure had no effect. `At risk' genotypes with higher incidence of CA could not be identified at this stage of the ongoing study. Conclusion: the challenge assay is able to detect enhanced susceptibility for CR caused by genetic predisposition for DNA repair deficiency. Our data indicate that environmental or occupational exposure to certain substances can interfere with DNA repair processes. As the process of induction of CR is associated with carcinogenesis, the challenge assay may provide a valuable biomarker for cancer epidemiology studies.