Chidamide alleviates TGF-β-induced epithelial-mesenchymal transition in lung cancer cell lines

Mol Biol Rep. 2016 Jul;43(7):687-95. doi: 10.1007/s11033-016-4005-z. Epub 2016 May 17.

Abstract

Transforming growth factor-β (TGF-β)-induced epithelial-mesenchymal transition is a critical process in the initiation of metastasis of various types of cancer. Chidamide is a class I histone deacetylase inhibitor with anti-tumor activity. This study investigated the effects of chidamide on TGF-β-mediated suppression of E-cadherin expression in adenocarcinomic lung epithelial cells and the molecular mechanisms involved in these effects. Western blot analysis, confocal microscopy, Quantitative methyl-specific PCR and bisulfite sequencing were used to evaluate the effects of different treatments on chidamide ameliorating TGF-β induced-E-cadherin loss. H3 acetylation binding to the promoter of E-cadherin was detected by chromatin immunoprecipitations (CHIP). We found that chidamide reduced the level of lung cancer cell migration observed using a Boyden chamber assay (as an indicator of metastatic potential). Chidamide inhibited TGF-β-induced SMAD2 phosphorylation and attenuated TGF-β-induced loss of E-cadherin expression in lung cancer cells by Western blotting and confocal microscopy, respectively. Quantitative methyl-specific PCR and bisulfite sequencing revealed that TGF-β-enhanced E-cadherin promoter methylation was ameliorated in cells treated with chidamide. We demonstrated that histone H3 deacetylation within the E-cadherin promoter was required for TGF-β-induced E-cadherin loss; cell treatment with chidamide increased the H3 acetylation detected by CHIP. Taken together, our results demonstrate that TGF-β suppressed E-cadherin expression by regulating promoter methylation and histone H3 acetylation. Chidamide significantly enhanced E-cadherin expression in TGF-β-treated cells and inhibited lung cancer cell migration. These findings indicate that chidamide has a potential therapeutic use due to its capacity to prevent cancer cell metastasis.

Keywords: Chidamide; Epithelial–mesenchymal transition; TGF-β.

MeSH terms

  • A549 Cells
  • Aminopyridines / pharmacology*
  • Antigens, CD
  • Antineoplastic Agents / pharmacology*
  • Benzamides / pharmacology*
  • Cadherins / genetics
  • Cadherins / metabolism
  • DNA Methylation
  • Drug Screening Assays, Antitumor
  • Epigenesis, Genetic
  • Epithelial-Mesenchymal Transition / drug effects*
  • Gene Expression
  • Gene Expression Regulation, Neoplastic
  • Histone Deacetylase Inhibitors / pharmacology*
  • Humans
  • Lung Neoplasms / drug therapy
  • Lung Neoplasms / pathology
  • Phosphorylation
  • Promoter Regions, Genetic
  • Protein Processing, Post-Translational
  • Transforming Growth Factor beta / physiology*

Substances

  • Aminopyridines
  • Antigens, CD
  • Antineoplastic Agents
  • Benzamides
  • CDH1 protein, human
  • Cadherins
  • Histone Deacetylase Inhibitors
  • Transforming Growth Factor beta
  • N-(2-amino-5-fluorobenzyl)-4-(N-(pyridine-3-acrylyl)aminomethyl)benzamide