Perturbation of Cellular Redox Homeostasis Dictates Divergent Effects of Polybutyl Cyanoacrylate (PBCA) Nanoparticles on Autophagy
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Nanoparticle Synthesis
2.3. Cells and Treatments
2.4. Immunoblotting
2.5. Microscopy
2.6. Inducible mKeima Assay System
2.6.1. Generation of Stable Cell Lines
2.6.2. Detection of mKeima Processing
2.7. Long-Lived Protein Degradation Assay (LLPD)
2.8. Autophagic LDH Sequestration Assay
2.9. Detection of Reactive Oxygen Species (ROS)
2.10. Glutathione Measurement (GSH)
2.11. Statistical Analysis
3. Results
3.1. PBCA NPs Trigger Concentration-Dependent, Divergent Effects on Autophagy
3.2. High Concentrations of PBCA Inhibit Autophagic LC3 Flux and LC3 Lipidation, Whereas Low Concentrations Activate Canonical, Yet LC3 Flux-Independent Autophagy
3.3. PBCA-Induced Redox Imbalance Is Required for Activation of Autophagy at Low PBCA Concentrations, and Mediates Inhibition of LC3 Lipidation and Autophagy, as Well as Subsequent Cytotoxicity at High PBCA Concentrations
3.4. H2O2 Inhibits LC3 Lipidation and Autophagic Degradation
3.5. PBCA-Induced Redox Imbalance Induces Phosphorylation of Beclin-1 and Bcl-2 via p38 and JNK
3.6. PBCA Activates Autophagy Downstream of p38 and JNK Activation
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sønstevold, T.; Engedal, N.; Torgersen, M.L. Perturbation of Cellular Redox Homeostasis Dictates Divergent Effects of Polybutyl Cyanoacrylate (PBCA) Nanoparticles on Autophagy. Cells 2021, 10, 3432. https://doi.org/10.3390/cells10123432
Sønstevold T, Engedal N, Torgersen ML. Perturbation of Cellular Redox Homeostasis Dictates Divergent Effects of Polybutyl Cyanoacrylate (PBCA) Nanoparticles on Autophagy. Cells. 2021; 10(12):3432. https://doi.org/10.3390/cells10123432
Chicago/Turabian StyleSønstevold, Tonje, Nikolai Engedal, and Maria Lyngaas Torgersen. 2021. "Perturbation of Cellular Redox Homeostasis Dictates Divergent Effects of Polybutyl Cyanoacrylate (PBCA) Nanoparticles on Autophagy" Cells 10, no. 12: 3432. https://doi.org/10.3390/cells10123432