Lymphoblasts of women with BRCA1 mutations are deficient in cellular repair of 8,5'-Cyclopurine-2'-deoxynucleosides and 8-hydroxy-2'-deoxyguanosine.
Mutations in breast and ovarian cancer susceptibility genes BRCA1 and BRCA2 predispose women to a high risk of these cancers. Here, we show that lymphoblasts of women with BRCA1 mutations who had been diagnosed with breast cancer are deficient in the repair of some products of oxidative DNA damage, namely, 8-hydroxy-2'-deoxyguanosine and 8,5'-cyclopurine-2'-deoxynucleosides. Cultured lymphoblasts from 10 individuals with BRCA1 mutations and those from 5 control individuals were exposed to 5 Gy of ionizing radiation to induce oxidative DNA damage and then allowed to repair this damage. DNA samples isolated from these cells were analyzed by liquid chromatography/mass spectrometry and gas chromatography/mass spectrometry to measure 8-hydroxy-2'-deoxyguanosine, (5'-S)-8,5'-cyclo-2'-deoxyadenosine, (5'-R)-8,5'-cyclo-2'-deoxyguanosine, and (5'-S)-8,5'-cyclo-2'-deoxyguanosine. After irradiation and a subsequent period of repair, no significant accumulation of these lesions was observed in the DNA from control cells. In contrast, cells with BRCA1 mutations accumulated statistically significant levels of these lesions in their DNA, providing evidence of a deficiency in DNA repair. In addition, a commonly used breast tumor cell line exhibited the same effect when compared to a relevant control cell line. The data suggest that BRCA1 plays a role in cellular repair of oxidatively induced DNA lesions. The failure of cells with BRCA1 mutations to repair 8,5'-cyclopurine-2'-deoxynucleosides indicates the involvement of BRCA1 in nucleotide-excision repair of oxidative DNA damage. This work suggest that accumulation of these lesions may lead to a high rate of mutations and to deleterious changes in gene expression, increasing breast cancer risk and contributing to breast carcinogenesis.
Rodriguez
Henry
H
Chemical Science and Technology Laboratory, and Statistical Engineering Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899-8311, USA.
Jaruga
Pawel
P
Leber
Dennis
D
Nyaga
Simon G
SG
Evans
Michele K
MK
Dizdaroglu
Miral
M
D000080242:8-Hydroxy-2'-Deoxyguanosine; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D004249:DNA Damage; D004260:DNA Repair / Q000235:genetics*; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000235:genetics; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry; D019398:Genes, BRCA1; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D009154:Mutation; D010084:Oxidation-Reduction; D011684:Purine Nucleosides / Q000235:genetics*
D011684:Purine Nucleosides; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine