Targeted Methylation of the Epithelial Cell Adhesion Molecule (EpCAM) Promoter to Silence Its Expression in Ovarian Cancer Cells

Abstract

The Epithelial Cell Adhesion Molecule (EpCAM) is overexpressed in many cancers including ovarian cancer and EpCAM overexpression correlates with decreased survival of patients. It was the aim of this study to achieve a targeted methylation of the EpCAM promoter and silence EpCAM gene expression using an engineered zinc finger protein that specifically binds the EpCAM promoter fused to the catalytic domain of the Dnmt3a DNA methyltransferase. We show that transient transfection of this construct increased the methylation of the EpCAM promoter in SKOV3 cells from 4–8% in untreated cells to 30%. Up to 48% methylation was observed in stable cell lines which express the chimeric methyltransferase. Control experiments confirmed that the methylation was dependent on the fusion of the Zinc finger and the methyltransferase domains and specific for the target region. The stable cell lines with methylated EpCAM promoter showed a 60–80% reduction of EpCAM expression as determined at mRNA and protein level and exhibited a significantly reduced cell proliferation. Our data indicate that targeted methylation of the EpCAM promoter could be an approach in the therapy of EpCAM overexpressing cancers.

DNA methylation profiling in human lung tissue identifies genes associated with COPD

“…Additionally, although we have observed correlation between DNA methylation and gene expression for the most robustly differentially methylated sites, we have not performed functional validation of these findings. Epigenome editing [36][37][38] is becoming a viable approach for validation of DNA methylation-expression correlations with refinements in laboratory methods including CRISPR-Cas9. 39 Application of these technologies to epigenetics research will accelerate the pace of discovery as well as provide a path to epigenetic therapeutics, and is a future direction for this work.…”

WITHDRAWN: Epigenetic Pathways Offer Targets for Ovarian Cancer Treatment

“…In fact, this is the case for the epithelial cell adhesion molecule (EpCAM) which is overexpressed in ovarian cancer and in other cancers as well. Nunna et al 17 achieved targeted methylation of the promoter of the gene, and this resulted in reduction in the expression of the gene, a mechanism which is very promising for cancer treatment.…”

Cancer induction and suppression with transcriptional control and epigenome editing technologies

“…The examinations of epigenome editing with DNA methylation/demethylation have been reported by many groups, where TET1 demethylase [36,41,58,59,61] and DNMT3A methyltransferase [31,32,41,43,44,[62][63][64] have been used as the epigenetic effectors.…”