Quantitative analysis of promoter methylation in exfoliated epithelial cells isolated from breast milk of healthy women.

TitleQuantitative analysis of promoter methylation in exfoliated epithelial cells isolated from breast milk of healthy women.
Publication TypeJournal Article
Year of Publication2010
AuthorsWong CM, Anderton DL, Smith-Schneider S, Wing MA, Greven MC, Arcaro KF
JournalEpigenetics : official journal of the DNA Methylation Society
Volume5
Issue7
Pagination645-55
Date Published2010 Oct 1
ISSN1559-2308
KeywordsAdult, Age Factors, Breast, Breast Neoplasms, Cadherins, CpG Islands, Cytoskeletal Proteins, DNA Methylation, Epithelial Cells, Female, Genes, Tumor Suppressor, Glutathione S-Transferase pi, Humans, Intercellular Signaling Peptides and Proteins, Lactation, Membrane Proteins, Middle Aged, Milk, Human, Promoter Regions, Genetic, Retinol-Binding Proteins, Cellular, Risk Factors, Tumor Suppressor Proteins, Young Adult
AbstractPromoter methylation analysis of genes frequently silenced in breast cancer is a promising indicator of breast cancer risk, as these methylation events are thought to occur long before presentation of disease. The numerous exfoliated epithelial cells present in breast milk may provide the breast epithelial DNA needed for detailed methylation analysis and assessment of breast cancer risk. Fresh breast milk samples and health, lifestyle, and reproductive history questionnaires were collected from 111 women. Pyrosequencing analysis was conducted on DNA isolated from the exfoliated epithelial cells immunomagnetically separated from the total cell population in the breast milk of 102 women. A total of 65 CpG sites were examined in six tumor suppressor genes: PYCARD (also known as ASC or TMS1), CDH1, GSTP1, RBP1 (also known as CRBP1), SFRP1, and RASSF1. A sufficient quantity of DNA was obtained for meaningful analysis of promoter methylation; women donated an average of 86 ml of milk with a mean yield of 32,700 epithelial cells per ml. Methylation scores were in general low as expected of benign tissue, but analysis of outlier methylation scores revealed a significant relationship between breast cancer risk, as indicated by previous biopsy, and methylation score for several CpG sites in CDH1, GSTP1, SFRP1, and RBP1. Methylation of RASSF1 was positively correlated with women’s age irrespective of her reproductive history. Promoter methylation patterns in DNA from breast milk epithelial cells can likely be used to assess breast cancer risk. Additional studies of women at high breast cancer risk are warranted.
DOI10.4161/epi.5.7.12961
Alternate JournalEpigenetics
PubMed ID20716965