Is 24-color FISH detection of in-vitro radiation-induced chromosomal aberrations suited to determine individual intrinsic radiosensitivity?
BACKGROUND
Reliable determination of intrinsic radiosensitivity in individual patients is a serious need in radiation oncology. Chromosomal aberrations are sensitive indicators of a previous exposure to ionizing irradiation. Former molecular cytogenetic studies showed that such aberrations as an equivalent of intrinsic radiosensitivity can be detected by fluorescence in-situ hybridization (FISH) techniques using whole chromosome painting (wcp) probes. However, only one up to three randomly chosen wcp probes have been applied for such approaches until now. As a random distribution of chromosomal rearrangements along the chromosomes is up to now still controversial, the power of the 24-color FISH approach should be elucidated in the present study.


METHODS
Lymphocytes derived from lymphoblastoid cell lines of one patient with Nijmegen breakage syndrome (NBS homozygote) and of two NBS heterozygotes and peripheral blood lymphocytes of two controls were analyzed. Samples of each patient/control were irradiated in vitro with 0.0 Gy, 0.7 Gy or 2.0 Gy prior to cultivation. Chromosomal aberrations were analyzed in detail and quantified by means of 24-color FISH as an expression of the individual intrinsic radiosensitivity.


RESULTS
24-color FISH analyses were done in a total of 1,674 metaphases. After in-vitro irradiation, 21% (0.7 Gy) or 57% (2.0 Gy) of the controls' cells, 15% (0.7 Gy) or 53% (2.0 Gy) of the heterozygotes' cells and 54% (0.7 Gy) or 79% (2.0 Gy) of the homozygote's cells contained aberrations. The highest average rates of breaks per mitosis [B/M] (0.7 Gy: 1.80 B/M, 2.0 Gy: 4.03 B/M) and complex chromosomal rearrangements [CCR] (0.7 Gy: 0.20 CCR/M, 2.0 Gy: 0.47 CCR/M) were observed in the NBS patient. Moreover, the proportion of different aberration types after irradiation showed a distinct increase in the rate of CCR combined with a decrease in dicentrics in the NBS homozygote.


CONCLUSIONS
To come to a more complete picture of radiation-induced aberrations and to detect and quantify genetically determined intrinsic radiosensitivity, a 24-color FISH approach using all human chromosome painting probes has been successfully applied on cytogenetic preparation lymphocytes. The controls and NBS heterozygotes were clearly distinguished from the NBS homozygote subject.
Kuechler
Alma
A
Department of Radiotherapy, Clinic of Radiology, Jena, Germany.
Neubauer
Susann
S
Grabenbauer
Gerhard G
GG
Claussen
Uwe
U
Liehr
Thomas
T
Sauer
Rolf
R
Wendt
Thomas G
TG
D000328:Adult; D002478:Cells, Cultured; D002648:Child; D002869:Chromosome Aberrations; D019457:Chromosome Breakage; D020223:Chromosome Painting; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008214:Lymphocytes / Q000528:radiation effects; D008297:Male; D008677:Metaphase; D008875:Middle Aged; D011829:Radiation Dosage; D011836:Radiation Tolerance; D013577:Syndrome