and only ~20% of induced mutations are of the deletion or rearrangement type (Miles and others 1990)—many deletions will have led to cell death. By contrast, radiation mutagenesis at the X-linked HPRT gene is much less constrained by neighboring sequence; induced mutation frequencies are substantially higher, and ~70% of induced mutations show HPRT deletion or rearrangement (Thacker 1986)—many more will have been tolerated (Bedford and Dewey 2002). Stated simply, gene loss mutations are characteristic of radiation, but their recovery in viable cells can be a major limiting factor. Also, gene amplification can result from the process of DSB repair (Difilippantonio and others 2002). As shown later, these features are important for consideration of carcinogenic mechanisms and are also discussed in respect of germline mutagenesis.
Deletion and rearrangement of APRT, HPRT, and other target genes do occur spontaneously but are generally less frequent than point mutation; in the case of most chemical mutagens, there is a strong bias toward the induction of point mutations (Thacker 1986; Miles and others 1990; Sankaranarayanan 1991).
Studies of the effect of radiation quality on the induction of gene mutations show a relationship similar between relative biological effectiveness (RBE) and LET to that noted for chromosome aberration induction. Mutagenic effectiveness peaks at a LET of 100–200 keV μm−1, with maximum RBE values usually in the range of 7–10 based largely on initial slopes of the dose-response (Cox and Masson 1979; Thacker and others 1979; Thacker 1992). Molecular analyses broadly suggest that a DNA deletion mechanism predominates for all radiation qualities (Thacker 1986; Gibbs and others 1987; Aghamohammadi and others 1992; Jostes and others 1994), but there are some conflicting data on this issue.
DNA sequence data for radiation-induced intragenic deletions in APRT and larger deletions encompassing HPRT indicate the frequent involvement of short direct or inverted DNA repeats at deletion breakpoints (Miles and others 1990; Morris and Thacker 1993). The presence of these short repeats is highly suggestive of an important role for illegitimate recombination processes in mutagenesis and, as for chromosome aberration induction, the involvement of DNA DSBs and error-prone NHEJ repair. Evidence for a close relationship between gene mutations and chromosome aberrations is that several induced gene mutations are associated with macroscopic region-specific chromosomal deletions or rearrangements (Cox and Masson 1978; Thacker and Cox 1983; Morris and Thacker 1993).
If, as molecular data suggest, error-prone NHEJ repair of DNA DSBs is the principal source of radiation-induced gene mutations, then a linear dose-response would be anticipated at low doses. For technical reasons, dose-response relationships for gene mutations are far less precise than those for chromosome aberrations. In general, however, a linear or linear-quadratic relationship provides a satisfactory description of the dose-response down to ~200 mGy (Thacker 1992) and, from limited data, at lower doses. The exceptions to this are the data from a particularly sensitive in vivo system that scores reversion mutations (as hair color changes) at the pink-eyed unstable (Bonassi and others 1995) locus in the mouse. Using this system, a linear nonthreshold low-LET dose response has been obtained at doses down to 10 mGy (Schiestl and others 1994), but as discussed later in this chapter, that system is probably reflecting a mutagenic component from the induction of genomic instability.
Studies of radiation-induced gene mutation in radiosensitive mutant cell lines indicate that increased mutability can be associated not only with defective repair of DNA DSB but also with processes that affect the regulation of DNA repair (Thacker and others 1994). Finally, in studies on the effects of low-dose-rate, low-LET radiation and other cellular repair-related factors (Thacker 1992), there is consistent evidence for potentially increased efficiency of repair of pre-mutagenic lesions at low dose rates, but none of these studies specifically suggest the presence of a low-dose threshold. The following sections consider specific aspects of cellular response relating to cell cycle effects, adaptive responses to radiation, the transfer of damage signals between cells (bystander effects), induced and persistent genomic instability, low-dose hyper-radiation sensitivity, and other aspects of dose-response.
Radiation-induced genomic instability has been defined as the manifestation of genetic damage in a certain fraction of irradiated cells over many cell cycles after they were irradiated (Little 2003). This persistent instability is expressed as chromosomal rearrangements, chromosomal bridge formation, chromatid breaks and gaps, and micronuclei (Grosovsky and others 1996; Murnane 1996; Poupon and others 1996; Limoli and others 1997a; Suzuki and others 1998) in the progeny of cells that survive irradiation. Reduction in cell cloning efficiency several generations after irradiation is called delayed lethality; it is supposedly a manifestation of genomic instability associated with an increase in lethal mutations (Seymour and Mothersill 1997). Also, gene mutations, such as HPRT mutations, that arise de novo several generations after irradiation are thought to be another manifestation of genomic instability. The spectrum of these de novo mutations resembles that of spontaneous mutations (i.e., primarily point mutations instead of deletions that are induced directly by irradiation; Little and others 1997). There is controversy, however, as to whether all of these different end points represent the same fundamental chromosomal alterations that result in genomic instability (Chang and Little 1992; Morgan and others 1996; Limoli and others 1997a; Little 1998; Mothersill and others 2000a). However, the similarity in the frequencies of genomic instability induced in X-irradiated cells, (3 to 19) × 10−5 per cell/mGy,