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

被引:5
作者
Failli, Alessandra [1 ]
Legitimo, Annalisa [1 ]
Migheli, Francesca [2 ]
Coppede, Fabio [2 ,3 ]
Mathers, John C. [4 ]
Spisni, Roberto [5 ]
Miccoli, Paolo [5 ]
Migliore, Lucia [2 ,3 ]
Consolini, Rita [1 ]
机构
[1] Univ Pisa, Dept Clin & Expt Med, I-56126 Pisa, Italy
[2] Univ Pisa, Div Med Genet, Dept Translat Res & New Technol Med & Surg, I-56126 Pisa, Italy
[3] Pisa Univ Hosp AOUP, Dept Lab Med, I-56126 Pisa, Italy
[4] Newcastle Univ, Inst Ageing & Hlth, Human Nutr Res Ctr, Newcastle Upon Tyne NE4 5PL, Tyne & Wear, England
[5] Univ Pisa, Dept Surg Med Mol & Crit Area Pathol, I-56124 Pisa, Italy
来源
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2014年 / 15卷 / 01期
关键词
epithelial cell adhesion molecule (EpCAM); immunomagnetic enrichment; epigenetics; colon cancer; DNA methylation; pyrosequencing; CIRCULATING TUMOR-CELLS; OVARIAN-CANCER; STEM-CELLS; EP-CAM; EXPRESSION; MICRODISSECTION; OVEREXPRESSION; EPIGENETICS; CARCINOMA; PROFILES;
D O I
10.3390/ijms15010044
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:44 / 57
页数:14
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