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Epigenetic effects of low-level sodium arsenite exposure on human liver HepaRG cells

Abstract

Chronic exposure to inorganic arsenic is associated with a variety of adverse health effects, including lung, bladder, kidney, and liver cancer. Several mechanisms have been proposed for arsenic-induced tumorigenesis; however, insufficient knowledge and many unanswered questions remain to explain the integrated molecular pathogenesis of arsenic carcinogenicity. In the present study, using non-tumorigenic human liver HepaRG cells, we investigated epigenetic alterations upon prolonged exposure to a noncytotoxic concentration of sodium arsenite (NaAsO2). We demonstrate that continuous exposure of HepaRG cells to 1 µM sodium arsenite (NaAsO2) for 14 days resulted in substantial cytosine DNA demethylation and hypermethylation across the genome, among which the claudin 14 (CLDN14) gene was hypermethylated and the most down-regulated gene. Another important finding was a profound loss of histone H3 lysine 36 (H3K36) trimethylation, which was accompanied by increased damage to genomic DNA and an elevated de novo mutation frequency. These results demonstrate that continuous exposure of HepaRG cells to a noncytotoxic concentration of NaAsO2 results in substantial epigenetic abnormalities accompanied by several carcinogenesis-related events, including induction of epithelial-to-mesenchymal transition, damage to DNA, inhibition of DNA repair genes, and induction of de novo mutations. Importantly, this study highlights the intimate mechanistic link and interplay between two fundamental cancer-associated events, epigenetic and genetic alterations, in arsenic-associated carcinogenesis.

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Acknowledgements

This work was partly supported by appointment of B. Borowa-Mazgaj to the Postgraduate Research Program and C.R. Steward to the Summer Student Research Program at the National Center for Toxicological Research administered by the Oak Ridge Institute for Science and Education. The views expressed in this manuscript do not necessarily represent those of the U.S. Food and Drug Administration.

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Correspondence to Igor P. Pogribny.

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Tryndyak, V.P., Borowa-Mazgaj, B., Steward, C.R. et al. Epigenetic effects of low-level sodium arsenite exposure on human liver HepaRG cells. Arch Toxicol 94, 3993–4005 (2020). https://doi.org/10.1007/s00204-020-02872-6

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  • DOI: https://doi.org/10.1007/s00204-020-02872-6

Keywords

  • Epigenetics
  • Sodium arsenite
  • Hepatotoxicity
  • HepaRG cells