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Knocking Down PIAS3 Reduces H2O2-induced Oxidative Stress Injury in HT22 Cells

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Abstract

Oxidative stress is involved in the pathological processes of many neurodegenerative diseases. Protein modification by small ubiquitin-like modifiers (SUMOs) has been implicated in oxidative stress injury. By conjugating SUMOs to their selective protein substrates, SUMO ligases play critical roles in regulating functions of proteins involved in oxidative stress injury. In this study, we screened siRNAs to knockdown the SUMO ligase PIAS3 to assess its role in H2O2-induced injury in HT22 cells. H2O2 stimulation increased total protein SUMOylation, facilitated intracellular reactive oxygen species (ROS) release, increased cleaved caspase-3 levels, promoted p38 and JNK activation (phosphorylation), upregulated apoptosis, and decreased cell viability. The siRNA against PIAS3 329-347 (siPIAS3-329) markedly downregulated the protein expression of PIAS3 and reversed these effects, whereas siNC (negative control) had no effect. Our findings demonstrate that PIAS3-mediated SUMOylation facilitates oxidative stress injury and p38/JNK-mediated cell apoptosis and that PIAS3 is a potential target to protect against oxidative stress injury.

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Abbreviations

SUMOs:

small ubiquitin-like modifiers

ROS:

reactive oxygen species

siPIAS3-329:

siRNA against PIAS3 329-347

siNC:

negative control siRNA

PIAS:

protein inhibitor of activated STAT

PBS:

phosphate-buffered saline

PI:

propidium iodide

SD:

standard deviation

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (32100769 and 82371401 to M.L. and 81100852 to C.-P.D.), the Natural Science Foundation of the Jiangsu Higher Education Institutions (20KJA310010 to C.-P.D.), and Xuzhou Medical University (D2020054 and JBGS202202 to M.L.).

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Authors and Affiliations

  1. Research Center for Biochemistry and Molecular Biology, Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical University, Xuzhou, Jiangsu, 221004, China

    Baixue Wang, Wenxin Qian, Kaiyue Chen, Meng Li & Caiping Du

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  1. Baixue Wang
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  2. Wenxin Qian
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  3. Kaiyue Chen
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  4. Meng Li
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Contributions

Baixue Wang performed biochemical research and analyzed data. Wenxin Qian and Kaiyue Chen assisted with cell culture and cell apoptotic detection. Meng Li and Caiping Du designed research, wrote the paper, and provided supervision and funding.

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Correspondence to Meng Li or Caiping Du.

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Wang, B., Qian, W., Chen, K. et al. Knocking Down PIAS3 Reduces H2O2-induced Oxidative Stress Injury in HT22 Cells. Cell Biochem Biophys (2024). https://doi.org/10.1007/s12013-024-01292-y

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