Transmission of gamma-ray-induced unstable chromosomal aberrations through successive mitotic divisions in human lymphocytes in vitro.

Abstract

Transmission of unstable chromosomal aberrations through successive mitotic divisions M1-M4 was analysed in 3 Gy gamma-ray-irradiated G0 human lymphocytes in vitro, harvested at 50, 72 and 96 h. Bromodeoxyuridine (BrdU) incorporation and subsequently the fluorescence plus Giemsa (FPG) method allowed the cell cycle status of each cell scored to be ascertained. Higher dicentric frequencies were observed in cells within the same post-irradiation division derived from extended culture times, indicating either a delay in cell cycle progression of aberrant cells or different lymphocyte sub-populations. The yield of complete dicentrics decreased by a factor of two in passing from M1 to M2 and showed further reductions of 26 and 44%, respectively, in moving from M2 to M3 and from M3 to M4. In the case of conventionally stained metaphases, wherein the cell cycle status does not enter the picture, the dicentric frequencies showed a reduction in yield of 39.6% at 72 h and 52.1% at 96 h compared with 50 h, because the cells analysed comprise a mixture of metaphases in different cell cycles. In the FPG stained first division metaphases, 92-100% of dicentrics analysed at 50, 72 and 96 h and in the conventionally stained metaphases, 90-94, 72-84 and 54-80% of dicentrics analysed at 50, 72 and 96 h respectively were complete dicentrics (with fragments). In the M1, M2, M3 and M4 metaphases analysed, 92-100, 50-89, 20-70 and 10-50% of dicentrics, respectively, were complete dicentrics and 55-75, 53-68, 43-57 and 36-50% excess acentrics, respectively, were seen in cells with complete dicentrics. Interindividual variation was observed in cell cycle kinetics, radiation-induced chromosomal aberrations and micronucleus frequencies. A salient feature of the delayed effects was the induction of giant cells and the mirror dicentrics observed in them. The reduction in dicentric frequencies due to either cell cycle delay or cell death in successive generations is aided by the multiplicity of aberrations. Bridge-breakage-fusion (BBF) events mediated by dicentric chromosomes may also be instrumental in the perpetuation of chromosomal instability.