Protective effect of survivin in doxorubicin-induced cell death in H9c2 cardiac myocytes

Beom Seob Lee, Soo Hyuk Kim, Taewon Jin, Eun Young Choi, Jaewon Oh, Sungha Park, Sang Hak Lee, Ji Hyung Chung, Seok Min Kang

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Background and Objectives: Apoptosis has been known to be an important mechanism of doxorubicin-induced cardiotoxicity. Survivin, which belongs to the inhibitor of apoptosis protein family, is associated with apoptosis and alteration of the cardiac myocyte molecular pathways. Therefore, we investigated the anti-apoptotic effect and cellular mechanisms of survivin using a protein delivery system in a doxorubicin-induced cardiac myocyte injury model. Materials and Methods: We constructed a recombinant survivin which was fused to the protein transduction domain derived from HIV-TAT protein. In cultured H9c2 cardiac myocytes, TAT-survivin (1 μM) was added for 1 hour prior to doxorubicin (1 μM) treatment for 24 hours. Cell viability and apoptosis were evaluated by 2-(4,5-dimethyltriazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, caspase-3 activity, and terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling assay. We measured the expression levels of several apoptosis-related signal proteins. Results: The survivin level was significantly reduced in a dose dependent manner up to 1 μM of doxorubicin in concentration. Purified recombinant TAT-survivin protein was efficiently delivered to H9c2 cardiac myocytes, and its transduction showed an anti-apoptotic effect, demonstrated by reduced caspase-3 activity and the apoptotic index, concomitantly with increased cell viability against doxorubicin injury. The phosphorylation of p38 mitogen-activated protein (MAP) kinase and the release of Smac from mitochondria were suppressed and the expression levels of Bcl-2 and cAMP response element-binding protein (CREB), the transcription factor of Bcl-2, were recovered following TAT-survivin transduction, indicating that survivin had an anti-apoptotic effect against doxorubicin injury. Conclusion: Our results suggest that survivin has a potentially cytoprotective effect against doxorubicin-induced cardiac myocyte apoptosis through mechanisms that involve a decrease in the phosphorylation of p38 MAP kinase, mitochondrial Smac release, and increased expression of Bcl-2 and CREB.

Original languageEnglish
Pages (from-to)400-407
Number of pages8
JournalKorean Circulation Journal
Volume43
Issue number6
DOIs
Publication statusPublished - 2013 Jun 1

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Cardiac Myocytes
Doxorubicin
Cell Death
Apoptosis
Cyclic AMP Response Element-Binding Protein
p38 Mitogen-Activated Protein Kinases
Caspase 3
Cell Survival
Wounds and Injuries
Phosphorylation
Inhibitor of Apoptosis Proteins
Human Immunodeficiency Virus Proteins
Proteins
DNA Nucleotidylexotransferase
Bromides
Mitochondria
Transcription Factors

All Science Journal Classification (ASJC) codes

  • Internal Medicine
  • Cardiology and Cardiovascular Medicine

Cite this

Lee, Beom Seob ; Kim, Soo Hyuk ; Jin, Taewon ; Choi, Eun Young ; Oh, Jaewon ; Park, Sungha ; Lee, Sang Hak ; Chung, Ji Hyung ; Kang, Seok Min. / Protective effect of survivin in doxorubicin-induced cell death in H9c2 cardiac myocytes. In: Korean Circulation Journal. 2013 ; Vol. 43, No. 6. pp. 400-407.
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abstract = "Background and Objectives: Apoptosis has been known to be an important mechanism of doxorubicin-induced cardiotoxicity. Survivin, which belongs to the inhibitor of apoptosis protein family, is associated with apoptosis and alteration of the cardiac myocyte molecular pathways. Therefore, we investigated the anti-apoptotic effect and cellular mechanisms of survivin using a protein delivery system in a doxorubicin-induced cardiac myocyte injury model. Materials and Methods: We constructed a recombinant survivin which was fused to the protein transduction domain derived from HIV-TAT protein. In cultured H9c2 cardiac myocytes, TAT-survivin (1 μM) was added for 1 hour prior to doxorubicin (1 μM) treatment for 24 hours. Cell viability and apoptosis were evaluated by 2-(4,5-dimethyltriazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, caspase-3 activity, and terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling assay. We measured the expression levels of several apoptosis-related signal proteins. Results: The survivin level was significantly reduced in a dose dependent manner up to 1 μM of doxorubicin in concentration. Purified recombinant TAT-survivin protein was efficiently delivered to H9c2 cardiac myocytes, and its transduction showed an anti-apoptotic effect, demonstrated by reduced caspase-3 activity and the apoptotic index, concomitantly with increased cell viability against doxorubicin injury. The phosphorylation of p38 mitogen-activated protein (MAP) kinase and the release of Smac from mitochondria were suppressed and the expression levels of Bcl-2 and cAMP response element-binding protein (CREB), the transcription factor of Bcl-2, were recovered following TAT-survivin transduction, indicating that survivin had an anti-apoptotic effect against doxorubicin injury. Conclusion: Our results suggest that survivin has a potentially cytoprotective effect against doxorubicin-induced cardiac myocyte apoptosis through mechanisms that involve a decrease in the phosphorylation of p38 MAP kinase, mitochondrial Smac release, and increased expression of Bcl-2 and CREB.",
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Protective effect of survivin in doxorubicin-induced cell death in H9c2 cardiac myocytes. / Lee, Beom Seob; Kim, Soo Hyuk; Jin, Taewon; Choi, Eun Young; Oh, Jaewon; Park, Sungha; Lee, Sang Hak; Chung, Ji Hyung; Kang, Seok Min.

In: Korean Circulation Journal, Vol. 43, No. 6, 01.06.2013, p. 400-407.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Protective effect of survivin in doxorubicin-induced cell death in H9c2 cardiac myocytes

AU - Lee, Beom Seob

AU - Kim, Soo Hyuk

AU - Jin, Taewon

AU - Choi, Eun Young

AU - Oh, Jaewon

AU - Park, Sungha

AU - Lee, Sang Hak

AU - Chung, Ji Hyung

AU - Kang, Seok Min

PY - 2013/6/1

Y1 - 2013/6/1

N2 - Background and Objectives: Apoptosis has been known to be an important mechanism of doxorubicin-induced cardiotoxicity. Survivin, which belongs to the inhibitor of apoptosis protein family, is associated with apoptosis and alteration of the cardiac myocyte molecular pathways. Therefore, we investigated the anti-apoptotic effect and cellular mechanisms of survivin using a protein delivery system in a doxorubicin-induced cardiac myocyte injury model. Materials and Methods: We constructed a recombinant survivin which was fused to the protein transduction domain derived from HIV-TAT protein. In cultured H9c2 cardiac myocytes, TAT-survivin (1 μM) was added for 1 hour prior to doxorubicin (1 μM) treatment for 24 hours. Cell viability and apoptosis were evaluated by 2-(4,5-dimethyltriazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, caspase-3 activity, and terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling assay. We measured the expression levels of several apoptosis-related signal proteins. Results: The survivin level was significantly reduced in a dose dependent manner up to 1 μM of doxorubicin in concentration. Purified recombinant TAT-survivin protein was efficiently delivered to H9c2 cardiac myocytes, and its transduction showed an anti-apoptotic effect, demonstrated by reduced caspase-3 activity and the apoptotic index, concomitantly with increased cell viability against doxorubicin injury. The phosphorylation of p38 mitogen-activated protein (MAP) kinase and the release of Smac from mitochondria were suppressed and the expression levels of Bcl-2 and cAMP response element-binding protein (CREB), the transcription factor of Bcl-2, were recovered following TAT-survivin transduction, indicating that survivin had an anti-apoptotic effect against doxorubicin injury. Conclusion: Our results suggest that survivin has a potentially cytoprotective effect against doxorubicin-induced cardiac myocyte apoptosis through mechanisms that involve a decrease in the phosphorylation of p38 MAP kinase, mitochondrial Smac release, and increased expression of Bcl-2 and CREB.

AB - Background and Objectives: Apoptosis has been known to be an important mechanism of doxorubicin-induced cardiotoxicity. Survivin, which belongs to the inhibitor of apoptosis protein family, is associated with apoptosis and alteration of the cardiac myocyte molecular pathways. Therefore, we investigated the anti-apoptotic effect and cellular mechanisms of survivin using a protein delivery system in a doxorubicin-induced cardiac myocyte injury model. Materials and Methods: We constructed a recombinant survivin which was fused to the protein transduction domain derived from HIV-TAT protein. In cultured H9c2 cardiac myocytes, TAT-survivin (1 μM) was added for 1 hour prior to doxorubicin (1 μM) treatment for 24 hours. Cell viability and apoptosis were evaluated by 2-(4,5-dimethyltriazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, caspase-3 activity, and terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling assay. We measured the expression levels of several apoptosis-related signal proteins. Results: The survivin level was significantly reduced in a dose dependent manner up to 1 μM of doxorubicin in concentration. Purified recombinant TAT-survivin protein was efficiently delivered to H9c2 cardiac myocytes, and its transduction showed an anti-apoptotic effect, demonstrated by reduced caspase-3 activity and the apoptotic index, concomitantly with increased cell viability against doxorubicin injury. The phosphorylation of p38 mitogen-activated protein (MAP) kinase and the release of Smac from mitochondria were suppressed and the expression levels of Bcl-2 and cAMP response element-binding protein (CREB), the transcription factor of Bcl-2, were recovered following TAT-survivin transduction, indicating that survivin had an anti-apoptotic effect against doxorubicin injury. Conclusion: Our results suggest that survivin has a potentially cytoprotective effect against doxorubicin-induced cardiac myocyte apoptosis through mechanisms that involve a decrease in the phosphorylation of p38 MAP kinase, mitochondrial Smac release, and increased expression of Bcl-2 and CREB.

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