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Title of Journal: Breast Cancer Res Treat

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Abbravation: Breast Cancer Research and Treatment

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Springer US

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DOI

10.1002/tea.3660070402

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1573-7217

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Promoter methylation reduces C/EBPEmphasis Type="

Authors: D Tang GS Sivko JW DeWille
Publish Date: 2005/12/02
Volume: 95, Issue: 2, Pages: 161-170
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Abstract

CCAAT/Enhancer Binding Proteins C/EBPs are a highly conserved family of leucine zipper proteins that regulate cell growth and differentiation C/EBPδ functions in the initiation and maintenance of mammary epithelial cell G0 growth arrest and ‘loss of function’ alterations in C/EBPδ gene expression have been reported in human breast cancer and in rodent carcinogeninduced mammary tumors The molecular mechanism underlying reduced C/EBPδ gene expression in mammary tumorigenesis however is unknown In this report we demonstrate that C/EBPδ gene expression is undetectable in the SUM52PE human breast cancer cell line and that silencing of SUM52PE C/EBPδ gene expression is due to epigenetic promoter hypermethylation 26/27 CpGs methylated The hypermethylated SUM52PE C/EBPδ gene promoter is associated with reduced levels of acetylated Histone H4 consistent with a closed transcriptionally inactive chromatin conformation Treatment with 5′azacytidine and trichostatin A TSA reactivates cytokineinduced SUM52PE C/EBPδ gene expression C/EBPδ gene expression is reduced to virtually undetectable levels in 32 18/57 of primary human breast tumors Sitespecific CpG methylation was observed in 33 6/18 of the low C/EBPδ expressing primary breast tumors CpG methylation adjacent to the C/EBPδ proximal promoter Sp1 site was associated with reduced C/EBPδ expression in a primary breast cancer sample Electromobility shift assays EMSA demonstrated a significant reduction in binding to oligos containing the CpG methylation 5′ to the Sp1 binding site These results demonstrate a direct link between C/EBPδ gene promoter hyper and site specificmethylation and reduced C/EBPδ gene expression in breast cancer cell lines and primary breast tumors

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