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Activation of AMP-Activated Protein Kinase Contributes to Doxorubicin-Induced Cell Death and Apoptosis in Cultured Myocardial H9c2 Cells

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Abstract

Despite its potent antitumor effect, clinical use of Doxorubicin is limited because of serious side effects including myocardial toxicity. Understanding the cellular mechanism involved in this process in a better manner is beneficial for optimizing Doxorubicin treatment. In the current study, the authors focus on the AMP-activated protein kinase (AMPK) in the said process. In this study, the authors discovered for the first time that Doxorubicin induces AMPK activation in cultured rat embryonic ventricular myocardial H9c2 cells. Reactive oxygen species (ROS)-dependent LKB1 activation serves as the upstream signal for AMPK activation by Doxorubicin. Evidence in support of the activation of AMPK contributing to Doxorubicin-induced H9c2 cell death/apoptosis—probably by modulating multiple downstream signal targets, including regulating JNK, p53, and inhibiting mTORC1—is provided in this article.

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Abbreviations

AMPK:

AMP-activated protein kinase

ROS:

Reactive oxygen species

Dox:

Doxorubicin

JNK:

c-Jun N-terminal kinases

mTOR:

Mammalian target of rapamycin

mTORC1:

mTOR complex 1

AICAR:

5′-Aminoimidazole-4-carboxamide ribonucleotide

TSC2:

Tuberous sclerosis protein 2

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

S6K1:

Ribosomal p70 S6 kinase

4E-BP1:

Eukaryotic initiation factor 4E (eIF4E)-binding protein 1

RNAi:

RNA interference

MnTBAP:

Mn(III)tetrakis (4-benzoic acid) porphyrin

NAC:

n-Acetyl-l-cysteine

FACS:

Fluorescence-activated cell sorting

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Acknowledgment

This study was supported by the Natural Science Foundation of Jiangsu Province (No. BK2010160).

Conflict of interest

None.

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Authors and Affiliations

  1. Department of Medical Oncology, Kunshan First People’s Hospital Affiliated to Jiangsu University, 91 Qianjin Road, Kunshan, 215300, Jiangsu Province, China

    Min-Bin Chen & Wen-Xiang Shen

  2. Department of Gastrointestinal Surgery, Kunshan First People’s Hospital Affiliated to Jiangsu University, 91 Qianjin Road, Kunshan, 215300, Jiangsu Province, China

    Xiao-Yang Wu

  3. Department of Medical Laboratory Center, Kunshan First People’s Hospital Affiliated to Jiangsu University, 91 Qianjin Road, Kunshan, 215300, Jiangsu Province, China

    Jin-Hua Gu

  4. Department of Surgery, Wuxi People’s Hospital of Nanjing Medical University, 299 Qingyang Road, Wuxi City, 214000, Jiangsu Province, China

    Qing-Tao Guo & Pei-Hua Lu

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  1. Min-Bin Chen
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  2. Xiao-Yang Wu
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  3. Jin-Hua Gu
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  4. Qing-Tao Guo
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  6. Pei-Hua Lu
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Corresponding author

Correspondence to Pei-Hua Lu.

Additional information

Min-Bin Chen and Xiao-Yang Wu contributed equally to this work.

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A

Effects of different concentration of target siRNA on AMPKα expression in H9c2 cells. H9c2 cells were either left un-transfected (non-transfected control), treated with transfection reagents or transfected with scramble siRNA or different concentration of AMPKα siRNA for 48 h using transfection method discussed above. AMPKα expression was detected by western blot; β-actin was detected as equal loading. AMPKα expression level was quantified after normalized toβ-actin. B, C Effects of scramble siRNA transfection on Doxorubicin-induced signal events in H9c2 cells. H9c2 cells were either left un-transfected (non-transfected control) or transfected with scramble siRNA (150 nM) for 48 h, followed by Doxorubicin (0.3 μg/ml) for indicated time points, LKB1, AMPK and JNK phosphorylation were detected by western blot (B). S6K phosphorylation, p53 expression and phosphorylation, Caspase 3 cleavage were also detected by western blot as demonstrated in (C) (EPS 2617 kb)

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Chen, MB., Wu, XY., Gu, JH. et al. Activation of AMP-Activated Protein Kinase Contributes to Doxorubicin-Induced Cell Death and Apoptosis in Cultured Myocardial H9c2 Cells. Cell Biochem Biophys 60, 311–322 (2011). https://doi.org/10.1007/s12013-011-9153-0

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