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A circular RNA from NFIX facilitates oxidative stress-induced H9c2 cells apoptosis

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Abstract

Myocardial infarction is the leading cause of death worldwide, and cardiomyocyte apoptosis during myocardial infarction and reperfusion is a significant factor of poor prognosis. As important regulatory molecules, biofunctions of circRNAs in the pathogenesis of myocardial infarction remain elusive. To confirm the expression level and biological function of circNFIX in cardiomyocytes upon oxidative stress. Divergent polymerase chain reaction and Sanger sequencing were performed to verify the circular structure. The stability of circNFIX was confirmed by RNase R treatment and actinomycin D assay. In order to simulate oxidative stress during myocardial infarction, H9c2 cells were subjected to hydrogen peroxide and hypoxia stimulation. In vivo, mouse models of myocardial ischemia were established. The biological function of circNFIX in cardiomyocytes was investigated through loss- and gain-of-function assays, and cardiomyocyte apoptosis level was detected by the terminal deoxyribonucleotidyl transferase-mediated TdT-mediated dUTP nick end labeling assay and Western blot. CircNFIX is abundant, conserved, and stable in H9c2 cells. The expression of circNFIX was significantly downregulated in cardiomyocytes subjected to oxidative stress. Enforced CircNFIX promotes H9c2 cells apoptosis induced by hydrogen peroxide, in sharp contrast to circNFIX knockdown. In this study, we found that circNFIX served as a pro-apoptosis factor in cardiomyocyte apoptosis. CircNFIX possesses potential to be the biomarker and therapeutic target in myocardial infarction.

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Funding

The support was provided by the National Natural Science Foundation of China (81770900), the Science and Technology Development Foundation of Shandong Province (2014GHY115025).

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  1. Xianglun Cui and Yanhan Dong contributed equally to this work.

Authors and Affiliations

  1. Department of Inspection, The Medical Faculty, Qingdao University, Qingdao, 266003, China

    Xianglun Cui, Mingyu Li, Xuekang Wang, Meiqing Jiang, Weiwei Yang, Gege Liu, Shuqi Sun & Wenhua Xu

  2. Institute for Translational Medicine, Qingdao University, Deng Zhou Road 38, Qingdao, 266021, China

    Yanhan Dong

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  1. Xianglun Cui
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Contributions

X. C. and Y. D. contributed equally to this study. W. X., X. C., and Y. D. contributed to the design of this study. X. C., Y. D., X. W., M. J., W. Y., G. L., and S. S. performed all the experiments. X. C. and M. L. performed the data collection and analysis. X. C. and Y. D. contributed to the interpretation of data and drafting the manuscript. All authors read and approved the final manuscript.

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Correspondence to Wenhua Xu.

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All institutional and national guidelines for the care and use of laboratory animals were followed and approved by the ethical standards of the Animal Care Committee, Qingdao University.

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Cui, X., Dong, Y., Li, M. et al. A circular RNA from NFIX facilitates oxidative stress-induced H9c2 cells apoptosis. In Vitro Cell.Dev.Biol.-Animal 56, 715–722 (2020). https://doi.org/10.1007/s11626-020-00476-z

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