Anthracycline-Induced Cardiac Injury Using a Cardiac Cell Line: Potential for Gene Therapy Studies

L’Ecuyer, Thomas; Horenstein, Mariano; Thomas, Ronald; Heide, Richard Vander

doi:10.1006/mgme.2001.3243

Cited by 24 publications

(15 citation statements)

References 30 publications

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“…Images are representative from three different cell preparations. Scale bar corresponds to 10 μm vitro studies using primary cardiomyocytes (Gabrielson et al 2007;Solem et al 1996;Zhou et al 2001) or other cell lines (L'Ecuyer et al 2001;Spallarossa et al 2004) have been widely reported in the literature. From the different models used, some conclusions are common to all: (a) Dox is activated at the mitochondrial level, forming a semiquinone radical, and (b) Dox causes oxidative stress in cardiac cells that particularly affects mitochondria (Berthiaume and Wallace 2007;Wallace 2003).…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2008

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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 Doxinduced 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.

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Section: Discussionmentioning

confidence: 99%

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

et al. 2008

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“…Wild-type (WT) and GST-expressing myocytes were plated in the absence of Tet and subjected to treatment with DOX (0.5 or 5 g/ml for 7 h) or vehicle. After the cells were rinsed, they were incubated for an additional 36 -48 h in fresh medium to allow the injury to evolve (25). Cells were collected by gentle trypsinization, followed by neutralizing with serum-containing medium.…”

Section: Methodsmentioning

confidence: 99%

“…We previously described the generation of multiple H9C2 cell lines stably transfected with the pTet-Off plasmid (25). Each transfected clone constitutively expresses a tetracycline transactivator protein at high level (15).…”

Section: Regulated Expression Of Gst By Stable Transfectantsmentioning

confidence: 99%

“…Oxidative stress precedes the development of irreversible cell injury after DOX exposure in H9C2 cells (25). To determine whether GST overexpression mitigated DOX-induced early oxidative stress, control cells were compared with GST-45 cells induced to maximally express GST.…”

Section: Regulated Expression Of Gst By Stable Transfectantsmentioning

confidence: 99%

“…We have recently developed a cell culture model of AC injury using cardiac-derived H9C2 myocytes, which differentiate and can express transfected foreign genes in a tightly regulated manner (25). We have suggested that this cell line represents a useful model in which to study the relationship between expression level of a putatively protective gene and the degree of protection from AC injury.…”

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confidence: 99%

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Glutathione S-transferase overexpression protects against anthracycline-induced H9C2 cell death

L’Ecuyer

Allebban

Thomas

et al. 2004

American Journal of Physiology-Heart and Circulatory Physiology

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Anthracyclines (AC) are antitumor antibiotics with significant activity against solid and hematologic malignancies. One problem preventing more widespread use has been the development of cardiac toxicity. Experimental evidence supports oxidant stress as an important trigger and/or mediator of AC-induced cardiotoxicity (ACT). Therefore, reducing oxidant stress should be protective against ACT. To determine whether antioxidant protein overexpression can reduce ACT, we developed a cell culture model system using the H9C2 cardiac cell line exhibiting controlled overexpression of the α4-isoform of glutathione- S-transferase (GST). Treatment with the AC doxorubicin (DOX) produced both oncosis, manifested by an increase in the number of cells staining positive for Trypan blue, and apoptosis, indicated by the presence of positive terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. In both cases, the loss of cell viability was preceded by an AC-induced increase in fluorescence with carboxy-2′,7′-dichlorofluorescein diacetate, demonstrating the presence of high levels of reactive oxygen species (ROS). The DOX-induced increase in ROS was reduced to control levels by maximal GST overexpression. Coincident with this elimination of oxidative stress, there was a reduction in both Trypan blue and TUNEL-positive cells, indicating that GST overexpression reduced both ROS and cell death in this model system. We conclude that GST overexpression may be an important part of a protective strategy against ACT and that this model system will aid in defining steps in the pathway(s) leading to AC-induced cell death that can be therapeutically manipulated.

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Induction of cellular glutathione-linked enzymes and catalase by the unique chemoprotective agent, 3H-1,2-dithiole-3-thione in rat cardiomyocytes affords protection against oxidative cell injury

Peng

2002

Pharmacological Research

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Anthracycline-Induced Cardiac Injury Using a Cardiac Cell Line: Potential for Gene Therapy Studies

Cited by 24 publications

References 30 publications

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

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

Glutathione S-transferase overexpression protects against anthracycline-induced H9C2 cell death

Induction of cellular glutathione-linked enzymes and catalase by the unique chemoprotective agent, 3H-1,2-dithiole-3-thione in rat cardiomyocytes affords protection against oxidative cell injury

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