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Int. J. Mol. Sci. 2014, 15(1), 44-57; doi:10.3390/ijms15010044
Article

Efficacy and Feasibility of the Epithelial Cell Adhesion Molecule (EpCAM) Immunomagnetic Cell Sorter for Studies of DNA Methylation in Colorectal Cancer

1,†
,
1,†
,
2
,
2,3
,
4
,
5
,
5
,
2,3
 and
1,*
1 Department of Clinical and Experimental Medicine, University of Pisa, Pisa 56126, Italy 2 Department of Translational Research and New Technologies in Medicine and Surgery, Division of Medical Genetics, University of Pisa, Pisa 56126, Italy 3 Department of Laboratory Medicine, Pisa University Hospital (AOUP), Pisa 56126, Italy 4 Human Nutrition Research Centre, Institute for Ageing & Health, Biomedical Research Building, Campus for Ageing & Vitality, Newcastle University, Newcastle upon Tyne NE4 5PL, UK 5 Department of Surgery, Medical, Molecular, and Critical Area Pathology, University of Pisa, Pisa 56124, Italy These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 7 November 2013 / Revised: 10 December 2013 / Accepted: 13 December 2013 / Published: 20 December 2013
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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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.
Keywords: DNA methylation; epigenetics; epithelial cell adhesion molecule (EpCAM); immunomagnetic enrichment; colon cancer; pyrosequencing DNA methylation; epigenetics; epithelial cell adhesion molecule (EpCAM); immunomagnetic enrichment; colon cancer; pyrosequencing
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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Failli, A.; Legitimo, A.; Migheli, F.; Coppedè, F.; Mathers, J.C.; Spisni, R.; Miccoli, P.; Migliore, L.; Consolini, R. Efficacy and Feasibility of the Epithelial Cell Adhesion Molecule (EpCAM) Immunomagnetic Cell Sorter for Studies of DNA Methylation in Colorectal Cancer. Int. J. Mol. Sci. 2014, 15, 44-57.

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