Structural alterations in breast stromal and epithelial DNA: the influence of 8,5'-cyclo-2'-deoxyadenosine.
(5'S)-8,5'-Cyclo-2'-deoxyadenosine (S-cdA), which arises from the reaction of the hydroxyl radical (*OH) with 2'-deoxyadenosine in DNA, is a lesion comprising a base-sugar linkage that distorts the DNA backbone. This structure impedes transcription and blocks polymerase action. Further, a single S-cdA lesion in the TATA box reduces gene expression. Considering the ability of S-cdA to disrupt DNA structure, which is likely associated with increased cancer risk, we determined S-cdA concentrations in the DNA of stroma, epithelium, and myoepithelium from normal breast tissues using liquid chromatography/mass spectrometry (LC/MS). We also identified differences in the base and backbone structures using Fourier transform-infrared (FT-IR) spectroscopy. LC/MS revealed that the lowest concentration of S-cdA in the stroma (0.04 +/- 0.02 lesions/10(6) bases) occurred in women ages 17 to 30. The highest concentration (0.13 +/- 0.07 lesions/10(6) bases) was found in women 33 to 46. FT-IR spectroscopy showed significant base and backbone differences in the stromal DNA between the women under 30 and those over 50. These findings imply that distortions in the geometry of the helix increase with age, reaching significant proportions in older women. No differences were found in the S-cdA concentrations between the three cell types, suggesting that the *OH attack on the base structure may be essentially random. Initial insight is provided on changes in DNA structure that potentially affect gene expression and increase breast cancer risk.
Anderson
Katie M
KM
Biochemical Oncology Program, Pacific Northwest Research Institute, Seattle, Washington, USA.
Jaruga
Pawel
P
Ramsey
Callie R
CR
Gilman
Naomi K
NK
Green
Virginia M
VM
Rostad
Steven W
SW
Emerman
Joanne T
JT
Dizdaroglu
Miral
M
Malins
Donald C
DC
D000293:Adolescent; D000328:Adult; D000367:Age Factors; D001940:Breast / Q000166:cytology*; D004247:DNA / Q000737:chemistry*; D003839:Deoxyadenosines / Q000032:analysis* / Q000494:pharmacology*; D004847:Epithelial Cells / Q000737:chemistry*; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D008875:Middle Aged; D009690:Nucleic Acid Conformation / Q000187:drug effects; D017154:Stromal Cells / Q000737:chemistry*
D003839:Deoxyadenosines; C056752:8,5'-cyclo-2'-deoxyadenosine; D004247:DNA