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Genome-wide analysis of tunicamycin-induced endoplasmic reticulum stress response and the protective effect of endoplasmic reticulum inhibitors in neonatal rat cardiomyocytes

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Abstract

Tunicamycin (TM) is an inducer of endoplasmic reticulum (ER) stress. However, which genes related to ER stress was induced in cardiomyocytes on a genome-wide scale remains poorly understood. Salubrinal and its derivatives are ER stress inhibitors. However, the cellular protection mechanisms remain unresolved. Neonatal rat cardiomyocytes were cultured from ventricles of one-day-old Wistar rats. Cells were exposed to salubrinal, its derivatives (PP1-12, PP1-24) or vehicle followed by TM treatment at different times. Total RNA was isolated from cells for RNA-sequencing analysis. The expressions of 189, 182, 556, 860, and 1314 genes were changed in cells exposed to TM for 1, 3, 6, 12, and 24 h. Five well-known UPR genes (Hspa5, Hsp90b1, Calr, Ddit3, and Atf4) were significantly increased in a time-dependent manner. Six not well-known genes (Hyou1, Herpud1, Manf, Creld2, Sdf2l1, and Slc3a2) were highlighted to be involved in ER stress. Compared with TM-only treated cells, the expressions of 36 genes upregulated by TM and 74 genes downregulated by TM were reversed by salubrinal. In comparison, 121 genes upregulated by TM and 92 genes downregulated by TM were reversed by PP1-12. Most genes altered by salubrinal are in the category of transcription (1 h) and cell cycle (24 h). Most genes altered by PP1-12 are in the category of response to ER stress (3 h) and cell cycle (24 h). Our findings help elucidate the mechanism for TM treatment and may be useful for future drug screens involved in ER stress.

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Acknowledgments

The authors thank the Biodynamic Optical Imaging Center of Peking University for their assistance with single cell whole transcriptome sequencing. This study was funded by the Ministry Science Foundation of the Chinese People’s Liberation Army during the 12th Five-Year Plan Period (No. BWS12J048) and Major International Science and Technology Cooperation Projects (No. 2013DFA31170).

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Authors and Affiliations

  1. Department of Geriatric Cardiology, The General Hospital of Chinese People’s Liberation Army, NO. 28, Fuxing Road, Haidian District, Beijing, 100853, China

    Chun-Lei Liu, Yun-Yun He, Xin Li, Song Li & Kun-Lun He

  2. Beijing Institute of Pharmacology and Toxicology, NO. 27, Taiping Road, Haidian District, Beijing, 100850, China

    Wu Zhong & Song Li

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  1. Chun-Lei Liu
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  2. Wu Zhong
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  3. Yun-Yun He
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  4. Xin Li
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  6. Kun-Lun He
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Correspondence to Kun-Lun He.

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Liu, CL., Zhong, W., He, YY. et al. Genome-wide analysis of tunicamycin-induced endoplasmic reticulum stress response and the protective effect of endoplasmic reticulum inhibitors in neonatal rat cardiomyocytes. Mol Cell Biochem 413, 57–67 (2016). https://doi.org/10.1007/s11010-015-2639-0

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  • DOI: https://doi.org/10.1007/s11010-015-2639-0

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