Protective Effect of Survivin in Doxorubicin-Induced Cell Death in H9c2 Cardiac Myocytes.
10.4070/kcj.2013.43.6.400
- Author:
Beom Seob LEE
1
;
Soo Hyuk KIM
;
Taewon JIN
;
Eun Young CHOI
;
Jaewon OH
;
Sungha PARK
;
Sang Hak LEE
;
Ji Hyung CHUNG
;
Seok Min KANG
Author Information
1. Graduate Program in Science for Aging, Yonsei University, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Apoptosis;
Doxorubicin;
Myocytes, cardiac
- MeSH:
Apoptosis;
Caspase 3;
Cell Death;
Cell Survival;
Cyclic AMP Response Element-Binding Protein;
Doxorubicin;
Humans;
Inhibitor of Apoptosis Proteins;
Mitochondria;
Myocytes, Cardiac;
p38 Mitogen-Activated Protein Kinases;
Phosphorylation;
Phosphotransferases;
Transcription Factors
- From:Korean Circulation Journal
2013;43(6):400-407
- CountryRepublic of Korea
- Language:English
-
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 microM) was added for 1 hour prior to doxorubicin (1 microM) 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 microM 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.
