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Hatta, Mitsutoki; Miyake, Yuki; Uchida, Kunitoshi; Yamazaki, Jun
Biochemical and biophysical research communications, 02/2018, Letnik: 496, Številka: 2Journal Article
Epithelial-mesenchymal transition (EMT) is a biological event in which epithelial cells lose their polarity and cell–cell adhesions and concomitantly acquire mesenchymal traits, and is thought to play an important role in pathological processes such as wound healing and cancer progression. In this study, we evaluated transforming growth factor (TGF)-β1-treated human keratinocyte HaCaT cells as an in vitro model of EMT. HaCaT cells were changed into an elongated fibroblast-like morphology, which is indicative of EMT in response to TGF-β1. Phalloidin staining demonstrated the formation of actin stress fibers in TGF-β1-treated cells. Quantitative RT-PCR analysis revealed that TGF-β1 increased the mRNA levels of EMT transcription factors (SNAI2, TWIST1, and ZEB1) and mesenchymal markers (CDH2, VIM, and FN1), while it decreased the transcripts of epithelial phenotypic genes (CLDN1, OCLN, KRT5, KRT15, KRT13, and TGM1). Furthermore, we found that KRT13 was drastically suppressed through the reduction of RNA polymerase II occupancy of its promoter, which was accompanied by a decrease in active histone marks (H3K4me3 and H3K27ac) and an increase in a repressive mark (H3K27me3) during EMT. These findings indicate that the TGF-β1-induced EMT program regulates a subset of epithelial and mesenchymal marker genes, and that KRT13 is transcriptionally suppressed through the modulation of the chromatin state at the KRT13 promoter in HaCaT cells. •TGF-β1 induced mesenchymal-like morphology in HaCaT cells.•TGF-β1-induced EMT partially controlled the gene expression of EMT markers.•KRT13 was identified as a TGF-β1-responsive epithelial marker in HaCaT cells.•Chromatin state of the KRT13 promoter was epigenetically regulated during EMT.
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