Epigenomic alterations and gene expression profiles in respiratory epithelia exposed to cigarette smoke condensate

F. Liu, J. K. Killian, M. Yang, R. L. Walker, J. A. Hong, M. Zhang, S. Davis, Y. Zhang, M. Hussain, S. Xi, M. Rao, P. A. Meltzer, D. S. Schrump

Research output: Contribution to journalArticle

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Abstract

Limited information is available regarding epigenomic events mediating initiation and progression of tobacco-induced lung cancers. In this study, we established an in vitro system to examine epigenomic effects of cigarette smoke in respiratory epithelia. Normal human small airway epithelial cells and cdk-4/hTERT-immortalized human bronchial epithelial cells (HBEC) were cultured in normal media with or without cigarette smoke condensate (CSC) for up to 9 months under potentially relevant exposure conditions. Western blot analysis showed that CSC mediated dose-and time-dependent diminution of H4K16Ac and H4K20Me3, while increasing relative levels of H3K27Me3; these histone alterations coincided with decreased DNA methyltransferase 1 (DNMT1) and increased DNMT3b expression. Pyrosequencing and quantitative RT-PCR experiments revealed time-dependent hypomethylation of D4Z4, NBL2, and LINE-1 repetitive DNA sequences; up-regulation of H19, IGF2, MAGE-A1, and MAGE-A3; activation of Wnt signaling; and hypermethylation of tumor suppressor genes such as RASSF1A and RAR-Β, which are frequently silenced in human lung cancers. Array-based DNA methylation profiling identified additional novel DNA methylation targets in soft-agar clones derived from CSC-exposed HBEC; a CSC gene expression signature was also identified in these cells. Progressive genomic hypomethylation and locoregional DNA hypermethylation induced by CSC coincided with a dramatic increase in soft-agar clonogenicity. Collectively, these data indicate that cigarette smoke induces cancer-associated epigenomic alterations in cultured respiratory epithelia. This in vitro model may prove useful for delineating early epigenetic mechanisms regulating gene expression during pulmonary carcinogenesis.

Original languageEnglish
Pages (from-to)3650-3664
Number of pages15
JournalOncogene
Volume29
Issue number25
DOIs
Publication statusPublished - 24-06-2010

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Respiratory Mucosa
Transcriptome
Epigenomics
Smoke
Tobacco Products
Epithelial Cells
DNA Methylation
Agar
Lung Neoplasms
DNA Fingerprinting
Nucleic Acid Repetitive Sequences
DNA
Methyltransferases
Tumor Suppressor Genes
Histones
Tobacco
Carcinogenesis
Up-Regulation
Clone Cells
Western Blotting

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Liu, F., Killian, J. K., Yang, M., Walker, R. L., Hong, J. A., Zhang, M., ... Schrump, D. S. (2010). Epigenomic alterations and gene expression profiles in respiratory epithelia exposed to cigarette smoke condensate. Oncogene, 29(25), 3650-3664. https://doi.org/10.1038/onc.2010.129
Liu, F. ; Killian, J. K. ; Yang, M. ; Walker, R. L. ; Hong, J. A. ; Zhang, M. ; Davis, S. ; Zhang, Y. ; Hussain, M. ; Xi, S. ; Rao, M. ; Meltzer, P. A. ; Schrump, D. S. / Epigenomic alterations and gene expression profiles in respiratory epithelia exposed to cigarette smoke condensate. In: Oncogene. 2010 ; Vol. 29, No. 25. pp. 3650-3664.
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Liu, F, Killian, JK, Yang, M, Walker, RL, Hong, JA, Zhang, M, Davis, S, Zhang, Y, Hussain, M, Xi, S, Rao, M, Meltzer, PA & Schrump, DS 2010, 'Epigenomic alterations and gene expression profiles in respiratory epithelia exposed to cigarette smoke condensate', Oncogene, vol. 29, no. 25, pp. 3650-3664. https://doi.org/10.1038/onc.2010.129

Epigenomic alterations and gene expression profiles in respiratory epithelia exposed to cigarette smoke condensate. / Liu, F.; Killian, J. K.; Yang, M.; Walker, R. L.; Hong, J. A.; Zhang, M.; Davis, S.; Zhang, Y.; Hussain, M.; Xi, S.; Rao, M.; Meltzer, P. A.; Schrump, D. S.

In: Oncogene, Vol. 29, No. 25, 24.06.2010, p. 3650-3664.

Research output: Contribution to journalArticle

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AU - Liu, F.

AU - Killian, J. K.

AU - Yang, M.

AU - Walker, R. L.

AU - Hong, J. A.

AU - Zhang, M.

AU - Davis, S.

AU - Zhang, Y.

AU - Hussain, M.

AU - Xi, S.

AU - Rao, M.

AU - Meltzer, P. A.

AU - Schrump, D. S.

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N2 - Limited information is available regarding epigenomic events mediating initiation and progression of tobacco-induced lung cancers. In this study, we established an in vitro system to examine epigenomic effects of cigarette smoke in respiratory epithelia. Normal human small airway epithelial cells and cdk-4/hTERT-immortalized human bronchial epithelial cells (HBEC) were cultured in normal media with or without cigarette smoke condensate (CSC) for up to 9 months under potentially relevant exposure conditions. Western blot analysis showed that CSC mediated dose-and time-dependent diminution of H4K16Ac and H4K20Me3, while increasing relative levels of H3K27Me3; these histone alterations coincided with decreased DNA methyltransferase 1 (DNMT1) and increased DNMT3b expression. Pyrosequencing and quantitative RT-PCR experiments revealed time-dependent hypomethylation of D4Z4, NBL2, and LINE-1 repetitive DNA sequences; up-regulation of H19, IGF2, MAGE-A1, and MAGE-A3; activation of Wnt signaling; and hypermethylation of tumor suppressor genes such as RASSF1A and RAR-Β, which are frequently silenced in human lung cancers. Array-based DNA methylation profiling identified additional novel DNA methylation targets in soft-agar clones derived from CSC-exposed HBEC; a CSC gene expression signature was also identified in these cells. Progressive genomic hypomethylation and locoregional DNA hypermethylation induced by CSC coincided with a dramatic increase in soft-agar clonogenicity. Collectively, these data indicate that cigarette smoke induces cancer-associated epigenomic alterations in cultured respiratory epithelia. This in vitro model may prove useful for delineating early epigenetic mechanisms regulating gene expression during pulmonary carcinogenesis.

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