Primary fibroblasts from BRCA1 heterozygotes display an abnormal G1/S cell cycle checkpoint following UVA irradiation but show normal levels of micronuclei following oxidative stress or mitomycin C treatment.

PURPOSE: There is evidence to suggest that the breast cancer predisposing gene, BRCA1, is involved in cell cycle control and the response to damage but mouse brca1+/- heterozygotes have no distinctive phenotype. Here the response to the three forms of cellular stress was examined in primary human fibroblasts from individuals with a +/+ or +/- genotype for BRCA1. METHODS AND MATERIALS: Fibroblasts from individuals carrying mutations in the BRCA1 gene were compared with those from those wild-type for BRCA1 in their response to long wavelength uv (UVA), hydrogen peroxide, and mitomycin C (MMC). Cell cycle progression and micronucleus formation (MN) were used as end points. RESULTS: After UVA treatment there was no difference between +/- and +/+ cells in the initial fall in DNA synthetic activity (G(1) arrest) but the reentry into S-phase was restored at a faster rate in the BRCA1+/- cells after UVA exposure. Thus, for three normal (+/+) cell lines irradiated in monolayer, S-phase values averaged 15 +/- 3.7% 14 h post-UVA (1 x 10(5) J/m(2)), as compared with 35.7 +/- 1.9 (range) for two BRCA1(+/-) strains. Because a defective G(1)/S checkpoint in BRCA1 heterozygotes could lead to a greater proportion of S-phase cells with unrepaired DNA damage (strand breaks) and a resultant increase in chromosomal instability, the frequency of micronuclei induced by UVA was examined. Three normal (+/+) and three mutant (+/-) strains (two of which were used in the cell cycle experiments) produced mean micronuclei frequencies of 0.077 +/- 0.016 and 0.094 +/- 0.04/binucleate cell respectively (not statistically significant), 48 h after UVA exposure. No differences were found between BRCA1+/+ and +/- cells in MN formation after treatment with MMC or hydrogen peroxide. CONCLUSION: Our data suggest a defective G(1)/S checkpoint in cells from BRCA1 heterozygotes in response to UVA although this is not reflected in genomic instability as measured by micronuclei induction after oxidative stress or MMC treatment.

The dose rate of UVA treatment influences the cellular response of HaCaT keratinocytes.

The contribution of UV exposure to the etiology of skin cancer and photoaging is undisputed. However, the effect of altering the intensity or dose rate of UV, which varies considerably with geographical location, the time of day or year, and the use of sunscreens, is not understood. In this study, the effect of altering the dose rate of UVA was investigated in the immortalized human keratinocyte cell line, HaCaT. Lowering the dose rate of UVA resulted in increased cytotoxicity, which correlated with increases in both lipid peroxidation and DNA damage. Furthermore, exposure at low dose rate did not appear to reduce the ability of UVA to induce the phenomenon of persistent genomic instability. Pretreatment with the antioxidant vitamin E significantly protected against UVA dose-rate effects observed with respect to lipid peroxidation and survival. Additionally, cell populations irradiated at low dose rate exhibited a shift towards a more pro-oxidant state. Taken together, these observations suggest an oxidative stress mechanism is underlying the UVA dose-rate effect. This study demonstrates that dose rates must be included as a key factor when evaluating the biological effects of UVA, especially considering the concerns, which exist regarding the efficacy and photostability of sunscreens to UVA.