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A carrier-assisted ChIP-seq method for estrogen receptor-chromatin interactions from breast cancer core needle biopsy samples.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Zwart, Wilbert 
Koornstra, Rutger 
Wesseling, Jelle 
Rutgers, Emiel 
Linn, Sabine 

Abstract

BACKGROUND: The Estrogen Receptor alpha (ERα) is the key transcriptional regulator in luminal breast cancer and is therefore the main target for adjuvant treatment of this subtype. Luminal gene signatures are dictated by the transcriptional capacities of ERα, which are a direct consequence of the receptors binding preference at specific sites on the chromatin. The identification of ERα binding signatures on a genome-wide level has greatly enhanced our understanding of Estrogen Receptor biology in cell lines and tumours, but the technique has its limitations with respect to its applicability in limited amounts of tumour tissue. RESULTS: Here, we present a refinement of the ChIP-seq procedures to enable transcription factor mapping on limited amounts of tissue culture cells as well as from a limited amount of tumor tissue derived from core needle biopsies. Our approach uses a carrier that can be removed prior to DNA amplification and sequencing. CONCLUSION: We illustrate the applicability of this refined technology by mapping the ERα genome-wide chromatin binding landscape in core needle biopsy material from primary breast tumours. With this, our refined technology permits for a high-resolution transcription factor mapping even from clinical samples.

Description

Keywords

Biopsy, Large-Core Needle, Breast Neoplasms, Chromatin, Chromatin Immunoprecipitation, Estrogen Receptor alpha, Humans, MCF-7 Cells, Protein Binding, Sequence Analysis

Journal Title

BMC Genomics

Conference Name

Journal ISSN

1471-2164
1471-2164

Volume Title

14

Publisher

Springer Science and Business Media LLC
Sponsorship
European Research Council (242664)
Cancer Research UK (15602)