DNA methylation profiling of asbestos-treated MeT5A cell line reveals novel pathways implicated in asbestos response

Arch Toxicol. 2018 May;92(5):1785-1795. doi: 10.1007/s00204-018-2179-y. Epub 2018 Mar 9.

Abstract

Occupational and environmental asbestos exposure is the main determinant of malignant pleural mesothelioma (MPM), however, the mechanisms by which its fibres contribute to cell toxicity and transformation are not completely clear. Aberrant DNA methylation is a common event in cancer but epigenetic modifications involved specifically in MPM carcinogenesis need to be better clarified. To investigate asbestos-induced DNA methylation and gene expression changes, we treated Met5A mesothelial cells with different concentrations of crocidolite and chrysotile asbestos (0.5 ÷ 5.0 µg/cm2, 72 h incubation). Overall, we observed 243 and 302 differentially methylated CpGs (≥ 10%) between the asbestos dose at 5 µg/cm2 and untreated control, in chrysotile and crocidolite treatment, respectively. To examine the dose-response effect, Spearman's correlation test was performed and significant CpGs located in genes involved in migration/cell adhesion processes were identified in both treatments. Moreover, we found that both crocidolite and chrysotile exposure induced a significant up-regulation of CA9 and SRGN (log2 fold change > 1.5), previously reported as associated with a more aggressive MPM phenotype. However, we found no correlation between methylation and gene expression changes, except for a moderate significant inverse correlation at the promoter region of DKK1 (Spearman rho = - 1, P value = 0.02) after chrysotile exposure. These results describe for the first time the relationship between DNA methylation modifications and asbestos exposure. Our findings provide a basis to further explore and validate asbestos-induced DNA methylation changes, that could influence MPM carcinogenesis and possibly identifying new chemopreventive target.

Keywords: Asbestos; Chrysotile; Crocidolite; DNA methylation; Gene expression; Mesothelioma.

MeSH terms

  • Antigens, Neoplasm / genetics
  • Asbestos / chemistry
  • Asbestos / toxicity*
  • Asbestos, Crocidolite / administration & dosage
  • Asbestos, Crocidolite / toxicity
  • Asbestos, Serpentine / administration & dosage
  • Asbestos, Serpentine / toxicity
  • Carbonic Anhydrase IX / genetics
  • Cell Line
  • DNA Methylation / drug effects*
  • Dose-Response Relationship, Drug
  • Epithelial Cells / drug effects
  • Humans
  • Lung Neoplasms / chemically induced
  • Lung Neoplasms / genetics
  • Mesothelioma / chemically induced
  • Mesothelioma / genetics
  • Mesothelioma, Malignant
  • Real-Time Polymerase Chain Reaction
  • Transcriptome / drug effects*

Substances

  • Antigens, Neoplasm
  • Asbestos, Serpentine
  • Asbestos, Crocidolite
  • Asbestos
  • CA9 protein, human
  • Carbonic Anhydrase IX