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Danforth DN, Warner AC, Wangsa D, et al. An Improved Breast Epithelial Sampling Method for Molecular Profiling and Biomarker Analysis in Women at Risk for Breast Cancer. Breast Cancer (Auckl). 2015;9:31-40doi: 10.4137/BCBCR.S23577.
Danforth, D. N., Warner, A. C., Wangsa, D., Ried, T., Duelli, D., Filie, A. C., & Prindiville, S. A. (2015). An Improved Breast Epithelial Sampling Method for Molecular Profiling and Biomarker Analysis in Women at Risk for Breast Cancer. Breast cancer : basic and clinical research, 931-40. https://doi.org/10.4137/BCBCR.S23577
Danforth, David N, et al. "An Improved Breast Epithelial Sampling Method for Molecular Profiling and Biomarker Analysis in Women at Risk for Breast Cancer." Breast cancer : basic and clinical research vol. 9 (2015): 31-40. doi: https://doi.org/10.4137/BCBCR.S23577
Danforth DN, Warner AC, Wangsa D, Ried T, Duelli D, Filie AC, Prindiville SA. An Improved Breast Epithelial Sampling Method for Molecular Profiling and Biomarker Analysis in Women at Risk for Breast Cancer. Breast Cancer (Auckl). 2015 Jun 08;9:31-40. doi: 10.4137/BCBCR.S23577. eCollection 2015. PMID: 26078587; PMCID: PMC4462519.
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Breast Cancer (Auckl). 2015 Jun 08;9:31-40. doi: 10.4137/BCBCR.S23577. eCollection 2015.

An Improved Breast Epithelial Sampling Method for Molecular Profiling and Biomarker Analysis in Women at Risk for Breast Cancer.

Breast cancer : basic and clinical research

David N Danforth, Andrew C Warner, Darawalee Wangsa, Thomas Ried, Dominik Duelli, Armando C Filie, Sheila A Prindiville

Affiliations

  1. Surgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
  2. Pathology- Histotechnology Laboratory, Laboratory Animal Sciences Program, Leidos Biomedical Research, Inc, Frederick, MD, USA.
  3. Genetics Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
  4. Department of Cellular and Molecular Pharmacology, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA.
  5. Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
  6. Office of the Director, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.

PMID: 26078587 PMCID: PMC4462519 DOI: 10.4137/BCBCR.S23577

Abstract

BACKGROUND: There is a strong need to define the molecular changes in normal at-risk breast epithelium to identify biomarkers and new targets for breast cancer prevention and to develop a molecular signature for risk assessment. Improved methods of breast epithelial sampling are needed to promote whole-genome molecular profiling, increase ductal epithelial cell yield, and reduce sample cell heterogeneity.

METHODS: We developed an improved method of breast ductal sampling with ductal lavage through a 22-gauge catheter and collection of ductal samples with a microaspirator. Women at normal risk or increased risk for breast cancer were studied. Ductal epithelial samples were analyzed for cytopathologic changes, cellular yield, epithelial cell purity, quality and quantity of DNA and RNA, and use in multiple downstream molecular applications.

RESULTS: We studied 50 subjects, including 40 subjects at normal risk for breast cancer and 37 subjects with non-nipple aspirate fluid-yielding ducts. This method provided multiple 1.0 mL samples of high ductal epithelial cell content (median ≥8 samples per subject of ≥5,000 cells per sample) with 80%-100% epithelial cell purity. Extraction of a single intact ductal sample (fluid and cells) or the separate frozen cellular component provided DNA and RNA for multiple downstream studies, including quantitative reverse transcription- polymerase chain reaction (PCR) for microRNA, quantitative PCR for the human telomerase reverse transcriptase gene, whole-genome DNA amplification, and array comparative genomic hybridization analysis.

CONCLUSION: An improved breast epithelial sampling method has been developed, which should significantly expand the acquisition and biomarker analysis of breast ductal epithelium in women at risk for breast cancer.

Keywords: breast cancer; breast duct sampling; breast ductal epithelium; breast epithelial profiling; normal breast epithelium

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