title: The p63 Gene Is Regulated by Grainyhead-like 2 (GRHL2) through Reciprocal Feedback and Determines the Epithelial Phenotype in Human Keratinocytes.
abstract: In this study, we investigated the effects of p63 modulation in epithelial plasticity in human keratinocytes. The p63 isoforms ΔNp63α, ΔNp63β, and ΔNp63γ were ectopically expressed in normal human epidermal keratinocytes (NHEKs). The epithelial or mesenchymal state was determined by morphological changes and altered expression of various markers, e.g. fibronectin, E-Cadherin, and keratin 14. Overexpression of ΔNp63α and ΔNp63β but not ΔNp63γ isoforms led to morphological changes consistent with epithelial-mesenchymal transition (EMT). However, only ΔNp63α overexpression was able to maintain the morphological changes and molecular phenotype consistent with EMT. Interestingly, knockdown of all p63 isoforms by transfection of p63 siRNA also led to the EMT phenotype, further confirming the role of p63 in regulating the epithelial phenotype in NHEKs. EMT in NHKs accompanied loss of Grainyhead-Like 2 (GHRL2) and miR-200 family gene expression, both of which play crucial roles in determining the epithelial phenotype. Modulation of GRHL2 in NHKs also led to congruent changes in p63 expression. ChIP revealed direct GRHL2 binding to the p63 promoter. GRHL2 knockdown in NHK led to impaired binding of GRHL2 and changes in the histone marks consistent with p63 gene silencing. These data indicate the presence of a reciprocal feedback regulation between p63 and GRHL2 in NHEKs to regulate epithelial plasticity.
authors.affiliation: From the School of Dentistry.
authors.forename: Shebli
authors.initials: S
authors.lastname: Mehrazarin
authors.affiliation: From the School of Dentistry.
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.affiliation: From the School of Dentistry.
authors.forename: Ju-Eun
authors.initials: JE
authors.lastname: Oh
authors.affiliation: From the School of Dentistry.
authors.forename: Zi X
authors.initials: ZX
authors.lastname: Liu
authors.affiliation: From the School of Dentistry, the Kyung Hee University, School of Dentistry, Seoul, 130-701, Korea.
authors.forename: Kyung L
authors.initials: KL
authors.lastname: Kang
authors.affiliation: From the School of Dentistry, the Kyung Hee University, School of Dentistry, Seoul, 130-701, Korea.
authors.forename: Jin K
authors.initials: JK
authors.lastname: Yi
authors.affiliation: From the School of Dentistry, Jonsson Comprehensive Cancer Center, and.
authors.forename: Reuben H
authors.initials: RH
authors.lastname: Kim
authors.affiliation: From the School of Dentistry, Jonsson Comprehensive Cancer Center, and.
authors.forename: Ki-Hyuk
authors.initials: KH
authors.lastname: Shin
authors.affiliation: From the School of Dentistry, the Kyung Hee University, School of Dentistry, Seoul, 130-701, Korea Jonsson Comprehensive Cancer Center, and David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, California 90095 and.
authors.forename: No-Hee
authors.initials: NH
authors.lastname: Park
authors.affiliation: From the School of Dentistry, Jonsson Comprehensive Cancer Center, and mkang@dentistry.ucla.edu.
authors.forename: Mo K
authors.initials: MK
authors.lastname: Kang
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D025461:Feedback, Physiological; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D053547:Keratin-14 / Q000235:genetics / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D061251:Primary Cell Culture; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D020033:Protein Isoforms / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D015415:Biomarkers; C081876:CKAP4 protein, human; D015820:Cadherins; D004268:DNA-Binding Proteins; D005353:Fibronectins; C470830:GRHL2 protein, human; D006657:Histones; C507540:KRT14 protein, human; D053547:Keratin-14; C533896:MIRN200 microRNA, human; D008565:Membrane Proteins; D035683:MicroRNAs; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D014157:Transcription Factors
keywords: GRHL2; epithelial-mesenchymal transition (EMT); epithelium; gene regulation; keratinocyte; p63