Analysis of epigenetic regulation of hypoxia-induced epithelial–mesenchymal transition in cancer cells by quantitative chromatin immunoprecipitation of histone deacetylase 3 (HDAC3)

Jian Qiu Wang, Min Zu Wu, Kou Juey Wu*

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

4 Scopus citations

Abstract

Epigenetics plays a key role in gene expression control. Histone modifications including acetylation/ deacetylation or methylation/demethylation are major epigenetic mechanisms known to regulate epithelial–mesenchymal transition (EMT)-associated gene expression during hypoxia-induced cancer metastasis. Chromatin immunoprecipitation (ChIP) assay is a powerful tool for investigation of histone modification patterns of genes of interest. In this chapter, we describe a protocol that uses chromatin immunoprecipitation (ChIP) to analyze the epigenetic regulation of EMT marker genes by deacetylation of acetylated Histone 3 Lys 4 (H3K4Ac) under hypoxia in a head and neck cancer cell line FaDu cells. Not only a method of ChIP coupled by real-time quantitative PCR but also the detailed conditions are provided based on our previously published studies.

Original languageEnglish
Pages (from-to)23-29
Number of pages7
JournalMethods in Molecular Biology
Volume1436
DOIs
StatePublished - 2016
Externally publishedYes

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media New York 2016.

Keywords

  • Chromatin immunoprecipitation
  • Epithelial–mesenchymal transition
  • HDAC3
  • Histone modification
  • Hypoxia
  • Real-time qPCR

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