ported to result in excess chromatid instability (Watson and others 1996). However, on the basis of the data summarized below, the consequences of postirradiation chromatid instability of bone marrow cells for hematopoietic neoplasia remains somewhat doubtful.
Cytogenetic characterization of myeloid leukemia induced in the same mouse strain by α-particles, neutrons, and X-rays did not reveal evidence of the LET-dependent cytogenetic footprint of induced chromatid instability that might be expected from in vitro cellular studies with bone marrow cells (Bouffler and others 1996). In addition, the very high α-particle relative biological effectiveness (RBE) for induced genomic instability in bone marrow cells in culture (Kadhim and others 1992) is somewhat inconsistent with the low α-particle RBE suggested to apply to leukemogenic risk in vivo (Breckon and Cox 1990; UNSCEAR 2000b).
Early studies of this form of induced instability depended on in vitro irradiation. Studies with humans exposed in vivo to low- and high-LET radiation (Tawn and others 2000b; Whitehouse and Tawn 2001) have found no evidence of induced chromatid instability in hemopoietic cells. The same negative result was obtained experimentally in the CBA/H mouse strain (Bouffler and others 2001). However Watson and colleagues (2001) provided data that suggested variable expression of in vivo induced chromatid instability in the CBH/H mouse strain. Since CBH/H is a highly inbred strain, such variable expression of chromatid instability cannot be ascribed to genetic variation. Experimental factors may therefore be of considerable importance, and relevant to this are the data of Bouffler and colleagues (2001), which indicate the existence of confounding stress factors that may account for in vitro and in vivo differences in the apparent expression of such instability.
These in vivo observations cast considerable doubt on the relevance of radiation-induced chromatid instability for risk of lymphohematopoietic tumors. This view is strengthened by studies showing that the genetic determinants of induced chromatid instability in mouse bone marrow cells differ from those of susceptibility to induced lymphohematopoietic neoplasia (Boulton and others 2001). A similar degree of doubt has been expressed following reanalysis of genomic instability data (Nakanishi and others 1999, 2001) relating to myeloid leukemia arising in A-bomb survivors (Cox and Edwards 2002; Little 2002).
Differences in radiosensitivity and susceptibility to radiation induction of specific tumors among inbred mouse strains are well recognized, and there is good evidence that the BALB/c mouse is unusually sensitive to the induction of tissue injury and mammary tumors (Roderick 1963; Storer and others 1988); on these criteria the C57BL/6 mouse falls into the radioresistant category. Initial cytogenetic studies showed that mammary epithelial cells cultured from irradiated BALB/c mice persistently expressed substantially more chromatid aberrations during passage than those derived from irradiated C57BL/6 animals (Ponnaiya and others 1997; Ullrich and Ponnaiya 1998). In follow-up investigations, the chromatid instability phenotype of BALB/c was shown to be associated with a partial deficiency in the NHEJ repair protein DNA-dependent protein kinase catalytic subunit (DNA PKcs) together with compromised postirradiation DNA DSB repair (Okayasu and others 2000). This study, which included an intercomparison of inbred mouse strains, showed deficiency of DNA-PKcs and DNA DSB repair to be restricted to BALB/c suggesting genetic associations with persistent genomic instability and mammary tumor susceptibility. In accord with this, molecular genetic analyses showed BALB/c to carry a rare variant form of the gene (Prkdc) encoding DNA-PKcs, and subsequent analysis of recombinant mice provided strong evidence that variant Prkdc directly determined DNA-PKcs deficiency and postirradiation chromatid instability in mammary epithelial cells (Yu and others 2001). On the basis of these data it was proposed that induced genomic instability and mammary tumor susceptibility were genetically codetermined. Importantly, these investigations provide genetic evidence that deficiencies in the repair of DNA DSB, rather than as-yet-undefined epigenetic phenomena, are likely to determine persistent chromatid instability in this mouse. The question as to whether such instability is a primary causal element in mammary tumorigenesis or a secondary in vitro consequence of DNA repair deficiency and clonal growth selection remains to be resolved.
Recent studies have also suggested a linkage between DNA-PKcs and maintenance of functional telomeres (Bailey and others 2004a, 2004b). As noted elsewhere in this report, the products of telomere dysfunction are dicentric chromosomes created by end-to-end fusion and sister-chromatid fusions, both of which can be associated with breakage-fusion-bridge cycles. More recently, a second product of telomere dysfunction, fusions between telomeres and the ends of broken DNA strands (i.e., DNA DSBs), have been described. Since telomere-DSB fusions have properties that differ from both chromosomal end fusions and ordinary chromosome aberrations, such fusions offer a potentially important new mechanism for induction of instability. These fusions appear to occur only under conditions of telomere dysfunction resulting from defects in the NHEJ pathway (Bailey and others 1999; Mills and others 2004). This suggests that genomic instability as a mechanism in radiation-induced cancer may be limited to specific circumstances in which individuals harbor specific DNA-repair deficiencies.
Telomeric repeat sequences (Bertoni and others 1994) cap the ends of mammalian chromosomes and serve to protect against replicative erosion and chromosomal fusion; in nor-