DNA damage in breast epithelial cells : detection by the single-cell gel (comet) assay and induction by human mammary lipid extracts

The presence of DNA damage in primary cultures of human mammary epithelial cells (HMECs), and the ability of extracts of human mammary lipid to cause such damage, has been investigated. Lipid extracts, prepared by a solid-phase procedure, and HMECs were obtained from breast tissue removed from healt...

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Bibliographic Details
Published in:Carcinogenesis (New York) 1997-12, Vol.18 (12), p.2299-2305
Main Authors: MARTIN, F. L, VENITT, S, CARMICHAEL, P. L, CROFTON-SLEIGH, C, STONE, E. M, COLE, K. J, GUSTERSON, B. A, GROVER, P. L, PHILLIPS, D. H
Format: Article
Language:English
Subjects:
Adult
Biological and medical sciences
Breast - chemistry
Breast - cytology
Carcinogenesis, carcinogens and anticarcinogens
Cells, Cultured
Chemical agents
Cytarabine - pharmacology
DNA Damage - drug effects
DNA Repair - drug effects
Epithelial Cells
Female
Humans
Hydroxyurea - pharmacology
Lipids - toxicity
Medical sciences
Middle Aged
Tumors
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Summary:The presence of DNA damage in primary cultures of human mammary epithelial cells (HMECs), and the ability of extracts of human mammary lipid to cause such damage, has been investigated. Lipid extracts, prepared by a solid-phase procedure, and HMECs were obtained from breast tissue removed from healthy women (ages 18-50 years) who were resident in the UK and undergoing elective reduction mammoplasties. DNA single strand breaks (SSBs) were detected using the single-cell gel assay (comet assay) with alkaline electrophoresis (pH 12.3) and quantified by measuring comet tail length (CTL) (microm). Untreated HMECs and HMECs incubated (30 min, 37 degrees C) with a mammary lipid extract, with or without DNA-repair inhibitors hydroxyurea (HU) and cytosine arabinoside (ara-C), were examined. Ionizing radiation was used as a positive control. An active lipid extract gave a linear dose-response over the range 2.0-12.2 g equivalents. When MCL-5 cells, a line of metabolically-competent human lymphoblastoid cells, were used to compare the DNA-damaging properties of lipid extracts from six different donors, significant interindividual variations (median CTLs were 15.0, 53.5, 32.5,
ISSN:0143-3334
1460-2180
1460-2180
DOI:10.1093/carcin/18.12.2299