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HSP25 down-regulation enhanced p53 acetylation by dissociation of SIRT1 from p53 in doxorubicin-induced H9c2 cell apoptosis

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Cell Stress and Chaperones Aims and scope

Abstract

Heat shock proteins (HSPs) play important roles in cellular stress resistance. Previous reports had already suggested that HSP27 played multiple roles in preventing doxorubicin-induced cardiotoxicity. Although HSP25 might have biological functions similar to its human homolog HSP27, the mechanism of HSP25 is still unclear in doxorubicin-induced cardiomyocyte apoptosis. To investigate HSP25 biological function on doxorubicin-induced apoptosis, flow cytometry was employed to analyze cell apoptosis in over-expressing HSP25 H9c2 cells in presence of doxorubicin. Unexpectedly, the H9c2 cells of over-expressing HSP25 have no protective effect on doxorubicin-induced apoptosis. Moreover, no detectable interactions were detected by coimmunoprecipitation between HSP25 and cytochrome c, and HSP25 over-expression failed in preventing cytochrome c release induced by doxorubicin. However, down-regulation of endogenous HSP25 by a specific small hairpin RNA aggravates apoptosis in H9c2 cells. Subsequent studies found that HSP25, but not HSP90, HSP70, and HSP20, interacted with SIRT1. Knockdown of HSP25 decreased the interaction between SIRT1 and p53, leading to increased p53 acetylation on K379, up-regulated pro-apoptotic Bax protein expression, induced cytochrome c release, and triggered caspase-3 and caspase-9 activation. These findings indicated a novel mechanism by which HSP25 regulated p53 acetylation through dissociation of SIRT1 from p53 in doxorubicin-induced H9c2 cell apoptosis.

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Abbreviations

shRNA:

Small hairpin RNA

cDNA:

Complementary DNA

RNAi:

RNA interference

FITC:

Fluorescein isothiocyanate

HSPs:

Heat shock proteins

HSP25:

Heat shock protein 25

HSP27:

Heat shock protein 27

SIRT1:

Sirtuin 1

DOX:

Doxorubicin

GAPDH:

Glycolytic glyceraldehyde-3-phosphate dehydrogenase

VDAC:

Voltage-dependent anion channel

TBST:

Tris-buffered saline Tween

BSA:

Bovine serum albumin

FCS:

Fetal calf serum

DMEM:

Dulbecco’s modified Eagle’s medium

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Acknowledgments

This work was supported by grants from the Major National Basic Research Program of China (no. 2007CB512007), the National Natural Science Foundation of China (81100106; 81100212; 81300113), A Project Supported by Scientific Research Fund of Hunan Provincial Education Department (11C1094; 11C1095), and the National College Students’ Innovation and Entrepreneurship Training Program (201210555011).

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Authors and Affiliations

  1. Department of Pathophysiology, Xiangya School of Medicine, Central South University, 110 Xiangya Road, Changsha, 410008, Hunan, People’s Republic of China

    Chi Zhang, Shunlin Qu, Xing Wei, Yansheng Feng, Honglin Zhu, Kangkai Wang, Ke Liu, Meidong Liu, Huali Zhang & Xianzhong Xiao

  2. Institute of Cardiovascular Disease, Key Laboratory for Arteriosclerology of Hunan Province, University of South China, Hengyang, 421001, Hunan, People’s Republic of China

    Chi Zhang, Shunlin Qu, Xing Wei & Jia Deng

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  1. Chi Zhang
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  2. Shunlin Qu
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  3. Xing Wei
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  4. Yansheng Feng
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  8. Ke Liu
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  10. Huali Zhang
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  11. Xianzhong Xiao
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Correspondence to Xianzhong Xiao.

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Zhang, C., Qu, S., Wei, X. et al. HSP25 down-regulation enhanced p53 acetylation by dissociation of SIRT1 from p53 in doxorubicin-induced H9c2 cell apoptosis. Cell Stress and Chaperones 21, 251–260 (2016). https://doi.org/10.1007/s12192-015-0655-3

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  • DOI: https://doi.org/10.1007/s12192-015-0655-3

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