Cell Biology and Toxicology

, Volume 25, Issue 3, pp 227–243 | Cite as

Morphological alterations induced by doxorubicin on H9c2 myoblasts: nuclear, mitochondrial, and cytoskeletal targets

  • Vilma A. Sardão
  • Paulo J. Oliveira
  • Jon Holy
  • Catarina R. Oliveira
  • Kendall B. Wallace
Article

Abstract

Doxorubicin (Dox) is a very potent antineoplastic agent used against several types of cancer, despite a cumulative cardiomyopathy that reduces the therapeutic index for treatment. H9c2 myoblast cells have been used as an in vitro model to study biochemical alterations induced by Dox treatment on cardiomyocyte cells. Despite the extensive work already published, few data are available regarding morphological alterations of H9c2 cells during Dox treatment. The purpose of the present work was to evaluate Dox-induced morphological alterations in H9c2 myoblasts, focusing especially on the nuclei, mitochondria, and structural fibrous proteins. Treatment of H9c2 cell with low concentrations of Dox causes alterations in fibrous structural proteins including the nuclear lamina and sarcomeric cardiac myosin, as well as mitochondrial depolarization and fragmentation, membrane blebbing with cell shape changes, and phosphatidylserine externalization. For higher Dox concentrations, more profound alterations are evident, including nuclear swelling with disruption of nuclear membrane structure, mitochondrial swelling, and extensive cytoplasm vacuolization. The results obtained indicate that Dox causes morphological alterations in mitochondrial, nuclear, and fibrous protein structures in H9c2 cells, which are dependent on the drug concentration. Data obtained with the present study allow for a better characterization of the effects of Dox on H9c2 myoblasts, used as a model to study Dox-induced cardiotoxicity. The results obtained also provide new and previously unknown targets that can contribute to understand the mechanisms involved in the cardiotoxicity of Dox.

Keywords

Cytoskeleton Doxorubicin H9c2 myoblasts Mitochondria Nuclei 

Abbreviations

DIC

differential interference contrast

Dox

doxorubicin

LDH

lactate dehydrogenase

PS

phosphatidylserine

ROS

reactive oxygen species

TMRM

tetramethylrhodamine methyl ester

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Vilma A. Sardão
    • 1
  • Paulo J. Oliveira
    • 1
  • Jon Holy
    • 2
  • Catarina R. Oliveira
    • 3
  • Kendall B. Wallace
    • 4
  1. 1.Center for Neurosciences and Cellular Biology, Department of ZoologyUniversity of CoimbraCoimbraPortugal
  2. 2.Department of Anatomy, Microbiology and PathologyUniversity of Minnesota Medical SchoolDuluthUSA
  3. 3.Center for Neurosciences and Cellular Biology, Department of Biochemistry, Medical SchoolUniversity of CoimbraCoimbraPortugal
  4. 4.Department of Biochemistry and Molecular BiologyUniversity of Minnesota Medical SchoolDuluthUSA

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