An allelic variant at the ATM locus is implicated in breast cancer susceptibility.

We have tested a simple procedure, disease association by locus stratification, for identifying breast cancer patients with pathogenetic allelic variants at several candidate loci. The strategy was based on the assumption of epistatic interactions of the candidates. We analyzed 66 independent cases from sib pairs affected with breast cancer that had previously been collected during an investigation of pathogenetic-allele-sharing at the HRAS1 mini-satellite locus. An exon 24 polymorphism of ATM, substituting arginine for proline was associated with breast cancer in these cases with an overall odds ratio of 4.5 (95% confidence interval, 1.2-20.5, nominal p = 0.02, 2-tail Fisher exact test). In the presence of a rare HRAS1 allele, the odds ratio increased to 6.9 (95% CI, 1.2-38.3, p = 0.03). Thus, our procedure identified at least one allelic variant of ATM associated with breast cancer, and indicated that the ATM locus may interact with HRAS1.

Distinct mutation patterns of breast cancer-associated alleles of the HRAS1 minisatellite locus.

DNA sequence analysis of 130 alleles of the HRAS1 minisatellite has demonstrated that breast cancer-associated variants arise as a consequence of both replication errors and gene conversions. Unlike mutations at other variable number of tandem repeats (VNTRs), high-risk variants of the HRAS1 minisatellite do not demonstrate positional polarity. Instead, most mutations occur at three hotspots, with replication errors confined to one hotspot, gene conversions to a second and a mixed pattern of mutation at the third. DNA sequence analysis of 66 low-risk a1 alleles revealed no evidence for hypermutation. Therefore, while the HRAS1 minisatellite may serve as a reporter for a broad-based group of mutational mechanisms, these results are consistent with a direct pathogenetic contribution by high-risk alleles as the biological basis underlying cancer association of this VNTR.

Modulation of the rat alveolar macrophage respiratory burst by hydroperoxides is calcium dependent.

Sublethal concentrations of hydroperoxides (H2O2 or tert-butylhydroperoxide) produce a dual effect upon the respiratory burst of rat alveolar macrophages in which low concentrations (< 50 microM) enhance and higher concentrations (> 50 microM) produce inhibition (J. K. Murphy, et al., Free Radical. Biol. Med. 18, 37-45, 1995). These effects correlate with transient versus sustained elevation of [Ca2+]i caused by exposure to hydroperoxides prior to stimulation of the respiratory burst. In the present study changes in [Ca2+]i caused by exposure to sublethal levels of hydroperoxide were buffered by incubating macrophages with the acetoxy-methyl ester of BAPTA, an intracellular Ca2+ chelator. The enhancement of the phorbol ester-stimulated respiratory burst by tBOOH was abolished by BAPTA, while the inhibition was attenuated. Thus, the modulation by tBOOH appears to be largely dependent upon the changes in [Ca2+]i. Receptor mediated stimulation of the respiratory burst (ADP stimulation) involves release of Ca2+ from the inositol-1,4,5-triphosphate (IP3)-sensitive pool in the endoplasmic reticulum. Comparisons were made of the effects of thapsigargin (TG), an endoplasmic reticulum Ca-ATPase inhibitor, with tBOOH on release of intracellular Ca2+ and the respiratory burst. Treatment with TG did not affect changes in [Ca2+]i caused by tBOOH or vice versa. Although TG decreased the ADP-stimulated respiratory burst, it had no effect upon tBOOH modulation. Thus, the effect of tBOOH upon the respiratory burst is dependent upon the release of Ca2+ and the release of Ca2+ occurs from a non-IP3-dependent pool. This aberrant mimicry of normal signal transduction underlies oxidative modulation of the respiratory burst.