title: Efficacy and feasibility of the epithelial cell adhesion molecule (EpCAM) immunomagnetic cell sorter for studies of DNA methylation in colorectal cancer.
abstract: The aim of this work was to assess the impact on measurements of methylation of a panel of four cancer gene promoters of purifying tumor cells from colorectal tissue samples using the epithelial cell adhesion molecule (EpCAM)-immunomagnetic cell enrichment approach. We observed that, on average, methylation levels were higher in enriched cell fractions than in the whole tissue, but the difference was significant only for one out of four studied genes. In addition, there were strong correlations between methylation values for individual samples of whole tissue and the corresponding enriched cell fractions. Therefore, assays on whole tissue are likely to provide reliable estimates of tumor-specific methylation of cancer genes. However, tumor cell tissue separation using immunomagnetic beads could, in some cases, give a more accurate value of gene promoter methylation than the analysis of the whole cancer tissue, although relatively expensive and time-consuming. The efficacy and feasibility of the immunomagnetic cell sorting for methylation studies are discussed.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Failli
authors.forename: Annalisa
authors.initials: A
authors.lastname: Legitimo
authors.forename: Francesca
authors.initials: F
authors.lastname: Migheli
authors.forename: Fabio
authors.initials: F
authors.lastname: Coppedè
authors.forename: John C
authors.initials: JC
authors.lastname: Mathers
authors.forename: Roberto
authors.initials: R
authors.lastname: Spisni
authors.forename: Paolo
authors.initials: P
authors.lastname: Miccoli
authors.forename: Lucia
authors.initials: L
authors.lastname: Migliore
authors.affiliation: Department of Clinical and Experimental Medicine, University of Pisa, Pisa 56126, Italy. rita.consolini@med.unipi.it.
authors.forename: Rita
authors.initials: R
authors.lastname: Consolini
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000951:Antigens, Neoplasm / Q000276:immunology* / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000276:immunology* / Q000378:metabolism; D015179:Colorectal Neoplasms / Q000378:metabolism* / Q000473:pathology; D018899:CpG Islands; D019175:DNA Methylation; D000071858:Epithelial Cell Adhesion Molecule; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D018189:Immunomagnetic Separation; D008297:Male; D008875:Middle Aged; D009367:Neoplasm Staging; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; D000071858:Epithelial Cell Adhesion Molecule