Single-walled and multi-walled carbon nanotubes induce sequence-specific epigenetic alterations in 16 HBE cells
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Manosij Ghosh1,*, Deniz Öner1,*, Radu C. Duca1, Bram Bekaert3,4, Jeroen A.J. Vanoirbeek1, Lode Godderis1,2,# and Peter H.M. Hoet1,#
1KU Leuven, Department of Public Health and Primary Care, Centre Environment and Health, B-3000 Leuven, Belgium
2Idewe, External Service for Prevention and Protection at Work, B-3001 Heverlee, Belgium
3Forensic Biomedical Sciences, Department of Imaging and Pathology, KU Leuven, University of Leuven, Leuven, Belgium
4Department of Forensic Medicine, Laboratory of Forensic Genetics and Molecular Archaeology, University Hospitals Leuven, Leuven, Belgium
*These authors contributed equally to this work and are co-first authors
Peter H.M. Hoet, email: firstname.lastname@example.org
Keywords: nanotoxicology; MWCNT; SWCNT; epigenetics; DNA methylation
Received: September 19, 2017 Accepted: March 15, 2018 Published: April 17, 2018
Recent studies have identified carbon nanotube (CNT)-induced epigenetic changes as one of the key players in patho-physiological response. In the present study, we investigated whether CNT exposure is associated with epigenetic changes in human bronchial epithelial cells (16 HBE), in vitro. We focused on global DNA methylation, methylation of LINE-1 elements and promoter sequence of twelve functionally important genes (SKI, DNMT1, HDAC4, NPAT, ATM, BCL2L11, MAP3K10, PIK3R2, MYO1C, TCF3, FGFR 1 and AGRN). Additionally, we studied the influence of CNT exposure on miRNA expression. Using a LC-MS/MS method and pyrosequencing for LINE-1, we observed no significant changes in global DNA methylation (%) between the concentrations of multi-walled and single-walled CNT (MWCNT and SWCNT, respectively). Significant changes in sequence-specific methylation was observed in at least one CpG site for DNMT1 (SWCNT), HDAC4 (MWCNT), NPAT/ATM (MWCNT and SWCNT), MAP3K10 (MWCNT), PIK3R2 (MWCNT and SWCNT) and MYO1C (SWCNT). While changes in DNA methylation of the genes were relatively small, these changes were associated with changes in RNA expression, especially for MWCNT. However, the study did not reveal any significant alteration in the miRNA expression, associated with MWCNT and SWCNT exposure. Based on our results, mainly MWCNT influence DNA methylation and expression of the studied genes and could have significant impact on several critical cellular processes.
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