1. Epigenetic regulation of the epithelial mesenchymal transition induced by synergistic action of TNF-α and TGF-β in retinal pigment epithelial cells
- Author
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Atsushi Mukai, Hiroki Hatanaka, Eiko Ito, Junji Hamuro, Shigeru Kinoshita, Morio Ueno, and Chie Sotozono
- Subjects
0301 basic medicine ,Epithelial-Mesenchymal Transition ,Biophysics ,Retinal Pigment Epithelium ,Biochemistry ,Histone Deacetylases ,Epigenesis, Genetic ,03 medical and health sciences ,0302 clinical medicine ,Cell Movement ,Transforming Growth Factor beta ,Humans ,Epithelial–mesenchymal transition ,Epigenetics ,Molecular Biology ,Cells, Cultured ,Cell Proliferation ,biology ,Tumor Necrosis Factor-alpha ,Microarray analysis techniques ,Chemistry ,CD44 ,Drug Synergism ,Cell Biology ,Histone Deacetylase Inhibitors ,Blot ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer research ,biology.protein ,Tumor necrosis factor alpha ,Histone deacetylase ,Transforming growth factor - Abstract
To clarify the influence of tumor necrosis factor (TNF)-α on fibrotic phenotypes induced by transforming growth factor (TGF)-β in retinal pigment epithelial cells (RPECs) by epigenetic regulation. Human primary retinal pigment epithelial cells (RPECs including ARPE19) were used in cultures in the presence or absence of TNF-α and/or TGF-β2. RT2 Profiler™ (Qiagen) was used for PCR Array for fibrosis and epithelial mesenchymal transition (EMT). Microarray analysis by 3D gene DNA chip was outsourced to Toray Industries Inc. Quantification of histone acetyl transferase (HAT)-related and histone deacetylase (HDAC) related gene expression were also analyzed. HDAC and HAT activity was measured using an EpiQuik HDAC and HAT Activity/Inhibition Assay Kit (Epigentek). CD44, MMP-9, HAT, and HDAC in RPECs were analyzed by western blotting. Analysis of expression of the EMT/fibrosis related CD44 and MMP-9 phenotypes induced by TNF-α+TGF-β2 revealed four alterations in RPECs: 1) abolition of TGF-β2-induced α-SMA by TNF-α; 2) synergy between TNF-α+TGF-β2 for induction of CD44 and MMP-9 phenotypes 3) no inhibition of HDAC activity by either TNF-α or TGF-β2; and 4) significant inhibition of HAT activity by either TNF-α or TGF-β2, but no synergy. The HDAC activation through HAT inhibition by TNF-α+TGF-β was counteracted by HDAC inhibitors, leading to the inhibition of EMT/fibrosis. EMT/fibrotic CD44 and MMP-9 phenotypes were epigenetically upregulated by concerted action of TNF-α and TGF-β in RPECs. The intervention in epigenetic regulation may hold potential in preventing intraocular proliferative diseases.
- Published
- 2021