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Analysis of Epigenetic Regulation of Hypoxia-Induced Epithelial–Mesenchymal Transition in Cancer Cells by Quantitative Chromatin Immunoprecipitation of Histone Deacetylase 3 (HDAC3)

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Histone Deacetylases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1436))

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.

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Acknowledgment

This work was supported in part to J.Q.W. by National Natural Science Foundation of China [81301850]; Educational Commission of Zhejiang Province of China [Y201328812]; and to K.J.W. by Ministry of Science and Technology Summit grant [MOST 103-2745-B-039-001-ASP]; National Science Council Frontier grant [NSC102-2321-B-010-001]; center of excellence for cancer research at Taipei Veterans General Hospital [MOHW104-TDU-B-211-124-001]; and National Health Research Institutes [NHRI-EX104-10230SI].

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Authors and Affiliations

  1. Institute of Aging Research, Department of Basic Medical Science, School of Medicine, Hangzhou Normal University, Hangzhou, 310036, China

    Jian-Qiu Wang

  2. Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, CA, 92037, USA

    Min-Zu Wu

  3. Research Center for Tumor Medical Science, Graduate Institute of Cancer Biology, China Medical University, Taichung, 404, Taiwan

    Kou-Juey Wu

Authors
  1. Jian-Qiu Wang
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  2. Min-Zu Wu
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  3. Kou-Juey Wu
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Corresponding author

Correspondence to Kou-Juey Wu .

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Editors and Affiliations

  1. Boston University School of Medicine, Boston, Massachusetts, USA

    Sibaji Sarkar

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Wang, JQ., Wu, MZ., Wu, KJ. (2016). Analysis of Epigenetic Regulation of Hypoxia-Induced Epithelial–Mesenchymal Transition in Cancer Cells by Quantitative Chromatin Immunoprecipitation of Histone Deacetylase 3 (HDAC3). In: Sarkar, S. (eds) Histone Deacetylases. Methods in Molecular Biology, vol 1436. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3667-0_3

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  • DOI: https://doi.org/10.1007/978-1-4939-3667-0_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3665-6

  • Online ISBN: 978-1-4939-3667-0

  • eBook Packages: Springer Protocols

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