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Chloral hydrate decreases gap junction communication in rat liver epithelial cells

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Abstract

Gap junction communication (GJC) is involved in controlling cell proliferation and differentiation. Alterations in GJC are associated with carcinogenesis, but the mechanisms involved are unknown. Chloral hydrate (CH), a by-product of chlorine disinfection of water, is carcinogenic in mice, and we demonstrated that CH reduced GJC in a rat liver epithelial cell line (Clone 9). To examine the mechanism(s) by which CH inhibits GJC, Clone 9 cells treated with CH were examined using Western blot, real-time polymerase chain reaction, immunocytochemical, and dye-communication techniques. Treatment with CH (0.1–5 mM for 24 h) resulted in a dose-dependent inhibition of GJC as measured by Lucifer yellow dye transfer. Western blot analysis demonstrated expression of connexin (Cx) 43 and 26 in control cells and reduced expression of Cx 43 but not Cx 26 protein from 0.1 to 1 mM CH. CH treatment from 2.5 to 5 mM caused an apparent increase in expression of both connexins that was concomitant with a reduction in mRNA expression for both connexins. Similarly, with immunocytochemistry, a dose-dependent decrease in Cx 43 staining at sites of cell–cell contact was apparent in CH (0.5–5 mM)-treated cultures, whereas no Cx 26 staining was observed. Thus, Clone 9 cells contain two types of connexins but only one type of plasma membrane channel. Understanding of the regulation of connexin may shed light on mechanisms responsible for inhibition of GJC by chemical carcinogens.

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Acknowledgements

The authors wish to thank Prof. Dr. C. Dame, Dr. D. Hackam, and Dr. A. Richard for helpful comments during the preparation of this manuscript and anonymous reviewers for valuable contributions to enhance the manuscript during journal evaluation.

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The information in this document has been funded wholly (or in part) by the US Environmental Protection Agency. It has been subjected to review by the National Health and Environmental Effects Research Laboratory and approved for publication. Approval does not signify that the contents reflect the views of the Agency, nor does mention of trade names or commercial products constitute endorsement or recommendation for use.

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  1. Juan Zhang

    Present address: Department of Neonatology, Charité—Universitätsmedizin Berlin, Augustenburger Platz 1, 13353, Berlin, Germany

Authors and Affiliations

  1. National Research Council, Washington, DC, USA

    Juan Zhang

  2. National Health and Environmental Effects Research Laboratory, Environmental Carcinogenesis Division [B-105-03], US Environmental Protection Agency, Research Triangle Park, NC, 27711, USA

    Rachel D. Grindstaff, Sheau-Fung Thai, Michael Kohan & Carl F. Blackman

  3. Department of Cell Biology and Physiology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15261, USA

    Sandra A. Murray

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Zhang, J., Grindstaff, R.D., Thai, SF. et al. Chloral hydrate decreases gap junction communication in rat liver epithelial cells. Cell Biol Toxicol 27, 207–216 (2011). https://doi.org/10.1007/s10565-011-9182-x

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