Loss of Fer Jeopardizes Metabolic Plasticity and Mitochondrial Homeostasis in Lung and Breast Carcinoma Cells
Abstract
:1. Introduction
2. Results
2.1. Impaired Metabolic Plasticity of Fer-Deficient H358 Cells
2.2. Elevated ROS Levels, Activation of the DNA Damage Response and Jeopardized Survival of Metabolically Challenged H358ΔFer Cells
2.3. Impaired Mitochondrial Function in Fer-Deficient H358 Cells
2.4. Fer Sustains Mitochondrial Metabolic Homeostasis in H358 and MDA-MB-231 Cells
2.5. Down-Regulated Urea Cycle in Fer-Deficient H358ΔFer and MDA-MB-231ΔFer Cells
2.6. Absence of Fer Impedes the Development of H358 and MDA-MB-231 Xenografts in Mice
3. Discussion
4. Materials and Methods
4.1. Cell Cultures
4.2. Generation of Stable CRSPR-Cas9 Knockout Clones
4.3. Western Blot Analysis
4.4. Immuno-Cytostaining
4.5. Determination of Cell-Growth Curves
4.6. Flow-Cytometry Cell-Cycle Analysis
4.7. Cell Death Analysis using Annexin/PI Staining
4.8. Measurement of ROS Levels
4.9. Measurement of the Mitochondrial OXPHOS Complex I Activity
4.10. Determining Mitochondrial Respiration Profile using “Mito-Stress” Analysis
4.11. Measurement of the Mitochondrial Potential using TMRE Staining
4.12. Determining NAD+ and Aspartate Levels
4.13. Quantification of Cellular ATP, CTP, TTP, Lys, Pro and Taurine Levels
4.14. Animal Experiments
4.15. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Mehazri, L.; Shpungin, S.; Bel, S.; Nir, U. Loss of Fer Jeopardizes Metabolic Plasticity and Mitochondrial Homeostasis in Lung and Breast Carcinoma Cells. Int. J. Mol. Sci. 2021, 22, 3387. https://doi.org/10.3390/ijms22073387
Mehazri L, Shpungin S, Bel S, Nir U. Loss of Fer Jeopardizes Metabolic Plasticity and Mitochondrial Homeostasis in Lung and Breast Carcinoma Cells. International Journal of Molecular Sciences. 2021; 22(7):3387. https://doi.org/10.3390/ijms22073387
Chicago/Turabian StyleMehazri, Linoy, Sally Shpungin, Shai Bel, and Uri Nir. 2021. "Loss of Fer Jeopardizes Metabolic Plasticity and Mitochondrial Homeostasis in Lung and Breast Carcinoma Cells" International Journal of Molecular Sciences 22, no. 7: 3387. https://doi.org/10.3390/ijms22073387