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BIOMARKER ASSAYS IN NIPPLE APIRATE FLUID
Klein, P., E. Glaser, L. Grogan, M. Keane, S. Lipkowitz, P. Soballe, L Brooks, J. Jenkins, S. M. Steinberg, D M. DeMarini, AND I. Kirsch. BIOMARKER ASSAYS IN NIPPLE APIRATE FLUID. THE BREAST JOURNAL 7(6):378-387, (2001).
The noninvasive technique of nipple aspiration as a potential source of biomarkers of breast cancer risk was evaluated. The feasibility of performing mutagenesis assays, amplifying DNA and performing protein electrophoresis on nipple aspirate fluid was explored. A tool was developed to measure the level of discomfort, if any, from this procedure. Twenty-five healthy women (20 pre-menopausal, 5 postmenopausal) were enrolled. Fluid was obtained using a modified breast pump. Pre-menopausal women were scheduled for 4-6 weekly aspirations, and postmenopausal women 1-2. Mutagenesis assays were performed using the Salmonella (Ames) assay. DNA amplification of several microsatellite regions was carried out using the polymerase chain reaction (PCR). Protein was quantified and 2-dimensional protein electrophoresis performed. Overall, fluid was obtained from 80% of women, and the level of discomfort was minimal. Acid hydrolysis of one sample resulted in mutagenicity; all 6 nonhydrolyzed samples were not mutagenic. Ability to amplify DNA ranged from 34% to 96% depending upon length of the microsatellite region examined. Average protein concentration was 70 ug/ml. 2D-protein electrophoresis was successfully performed on samples from 2 subjects. Nipple aspiration is a simple technique, easily learned, and well tolerated, which yields a reagent useful for a variety of investigations. This technique may facilitate the identification and application of biomarkers for future breast cancer risk assessment and chemopreventive protocols.
Record Details:Record Type: DOCUMENT (JOURNAL/PEER REVIEWED JOURNAL)
Organization:U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
NATIONAL HEALTH AND ENVIRONMENTAL EFFECTS RESEARCH LABORATORY
ENVIRONMENTAL CARCINOGENESIS DIVISION
GENETIC AND CELLULAR TOXICOLOGY BRANCH