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Isoproterenol Cytotoxicity is Dependent on the Differentiation State of the Cardiomyoblast H9c2 Cell Line

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An Erratum to this article was published on 21 July 2011

Abstract

H9c2 cells are used as a surrogate for cardiac cells in several toxicological studies, which are usually performed with cells in their undifferentiated state, raising questions on the applicability of the results to adult cardiomyocytes. Since H9c2 myoblasts have the capacity to differentiate into skeletal and cardiac muscle cells under different conditions, the hypothesis of the present work was that cells in different differentiation states differ in their susceptibility to toxicants. In order to test the hypothesis, the effects of the cardiotoxicant isoproterenol (ISO) were investigated. The present work demonstrates that differentiated H9c2 cells are more susceptible to ISO toxicity. Cellular content of beta1-adrenergic receptors (AR), beta3-AR, and calcineurin is decreased as cells differentiate, as opposed to the content on the mitochondrial voltage-dependent anion channel (VDAC) and phosphorylated p38-MAPK, which increase. After ISO treatment, the pro-apoptotic protein Bax increases in all experimental groups, although only undifferentiated myoblasts up-regulate the anti-apoptotic Bcl-2. Calcineurin is decreased in differentiated H9c2 cells, which suggests an important role against ISO-induced cell death. The results indicate that the differentiation state of H9c2 myoblasts influence ISO toxicity, which may involve calcineurin, p38-MAPK, and Bax/Bcl-2 alterations. The data also provide new insights into cardiovascular toxicology during early development.

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Acknowledgments

We are very grateful to Dr. Kendall B. Wallace (University of Minnesota, Medical School, Duluth, USA) for useful discussions and for providing initial funding for this study. This work was supported by a research grant from the Portuguese Foundation for Science and Technology (FCT), reference PTDC/QUI/64358/2006, to Paulo J. Oliveira. Ana Branco, Carolina Moreira, Sandro Pereira, and Vilma Sardão are supported by fellowships from the FCT (SFRH/BD/41384/2007, SFRH/BD/33892/2009, SFRH/BD/37933/2007, and SFRH/BPD/31549/2006, respectively).

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Authors and Affiliations

  1. Center for Neuroscience and Cell Biology, Department of Life Sciences, University of Coimbra, 3004-517, Coimbra, Portugal

    Ana F. Branco, Sandro L. Pereira, Ana C. Moreira, Vilma A. Sardão & Paulo J. Oliveira

  2. Department of Anatomy, Microbiology and Pathology, University of Minnesota Medical School, Duluth, MN, USA

    Jon Holy

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  1. Ana F. Branco
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Correspondence to Paulo J. Oliveira.

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An erratum to this article can be found at http://dx.doi.org/10.1007/s12012-011-9131-1

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Branco, A.F., Pereira, S.L., Moreira, A.C. et al. Isoproterenol Cytotoxicity is Dependent on the Differentiation State of the Cardiomyoblast H9c2 Cell Line. Cardiovasc Toxicol 11, 191–203 (2011). https://doi.org/10.1007/s12012-011-9111-5

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