title: Analysis of Epigenetic Regulation of Hypoxia-Induced Epithelial-Mesenchymal Transition in Cancer Cells by Quantitative Chromatin Immunoprecipitation of Histone Deacetylase 3 (HDAC3).
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.
authors.affiliation: Institute of Aging Research, Department of Basic Medical Science, School of Medicine, Hangzhou Normal University, Hangzhou, 310036, China.
authors.forename: Jian-Qiu
authors.initials: JQ
authors.lastname: Wang
authors.affiliation: Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, CA, 92037, USA.
authors.forename: Min-Zu
authors.initials: MZ
authors.lastname: Wu
authors.affiliation: Research Center for Tumor Medical Science, Graduate Institute of Cancer Biology, China Medical University, Taichung, 404, Taiwan. wukj@mail.cmu.edu.tw.
authors.forename: Kou-Juey
authors.initials: KJ
authors.lastname: Wu
mesh_terms: D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D058750:Epithelial-Mesenchymal Transition; D015972:Gene Expression Regulation, Neoplastic; D053263:Gene Regulatory Networks; D006655:Histone Deacetylases / Q000378:metabolism*; D006801:Humans; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D000099186:Histone Deacetylase 3
chemical_list: D006655:Histone Deacetylases; D000099186:Histone Deacetylase 3
keywords: Chromatin immunoprecipitation; Epithelial–mesenchymal transition; HDAC3; Histone modification; Hypoxia; Real-time qPCR