Effects of Cardiotrophin-1 on Adriamycin-Induced Apoptosis in H9c2 Cardiomyoblasts.
10.4070/kcj.2008.38.5.264
- Author:
Jae Ok SHIN
1
;
Eun Seon JU
;
Hyun Mi SONG
;
Soo Hyeon YUN
;
Byung Kwan LIM
;
Jin Ho CHOI
;
Duk Kyung KIM
;
Eun Seok JEON
Author Information
1. Department of Medicine, Sungkyunkwan University School of Medicine, Cardiac and Vascular Center, Samsung Medical Center, Seoul, Korea. esjeon@smc.samsung.co.kr
- Publication Type:Original Article ; In Vitro
- Keywords:
Adriamycin;
Cardiotrophin-1;
Apoptosis;
Cell protection
- MeSH:
Apoptosis;
Caspase 3;
Cell Count;
Cell Death;
Cell Survival;
Cytoprotection;
Doxorubicin;
Heart;
Humans;
Myocytes, Cardiac;
Phosphorylation;
RNA, Messenger
- From:Korean Circulation Journal
2008;38(5):264-269
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND AND OBJECTIVES: Adriamycin (doxorubicin, ADR) is a highly effective anti-neoplastic drug, but its clinical use is limited by its adverse side effects on the heart. Cardiotrophin (CT-1), a potent cardiac survival factor, is capable of inhibiting apoptosis in cardiac myocytes. The aim of this study was to investigate the cyto-protective effects of CT-1 against ADR-induced apoptosis in vitro. MATERIALS AND METHODS: We determined a reasonable ADR concentration for inducing cell death by utilizing a cell survival test performed in a dose-dependent manner. To determine the requirements for apoptosis in ADR-treated cardiac myocytes (H9c2 cells), we examined the effect of CT-1 on survival and apoptotic changes using a cell counting kit (CCK), RT-PCR, and Western blotting. RESULTS: In analyzing cell survival as determined by CCK, ADR-induced cell death was found to occur in a dose-dependent manner (50% death at 24 hours after 2 micrometer of ADR), and ADR was shown to decrease procaspase-3. On RT-PCR, expression of Bax-alpha mRNA increased and Bcl-2 decreased during the 24 hours after ADR treatment. Consequently, the ratio of Bax-alpha/Bcl-2 mRNA peaked at 24 hours after ADR treatment. In contrast, CT-1 effectively attenuated the ADR-induced cell death in a dose-dependent manner. The changes in Bax-alpha and Bcl-2 mRNA expression after ADR treatment were reversed by CT-1 (1 ng/mL) treatment. The protein levels of procaspase-3 decreased after ADR treatment, an effect which was reversed by CT-1 treatment. Akt phosphorylation was also increased by CT-1, demonstrating that CT-1 inhibited apoptosis induced by ADR. CONCLUSION: These data demonstrated that ADR-induced apoptosis of cardiomyocytes can be prevented by CT-1; therefore, it may be possible to use CT-1 as a cardioprotective agent during ADR chemotherapy in patients with cancer.
