GLUL Ablation Can Confer Drug Resistance to Cancer Cells via a Malate-Aspartate Shuttle-Mediated Mechanism
Abstract
:1. Introduction
2. Results
2.1. Transient GLUL Knockdown Induces Drug Resistance
2.2. GLUL Knockout Cells Display Drug Resistance
2.3. GLUL KO-Associated Drug Resistance Correlates with Apoptosis
2.4. GLUL Knockout Cells Display Glucose Dependence
2.5. A Metabolic Phenotype Involving the Malate-Aspartate Shuttle Supports Drug Resistance Associated with GLUL KO
2.6. Inhibition of the Malate-Aspartate Shuttle Reveals Metabolic Vulnerability Associated with Resistance
2.7. Rescuing GLUL Expression Sensitizes A549 KO Cells to Drugs
3. Discussion
4. Material and Methods
4.1. Cell Culture and Reagents
4.2. GLUL siRNA Transfection
4.3. Establishment of Inducible GLUL KD Cell Lines by shRNA Expression
4.4. Establishment of Stable GLUL KO Cell Lines by CRISPR/Cas9
4.5. GLUL Re-Expression in A549 GLUL KO Cells
4.6. MTS Assay and Western Blot Analysis
4.7. Clonogenic Assay
4.8. Flow Cytometry (FACS)-Based Apoptosis Analysis
4.9. Metabolic Profiling and Stable Isotope Labeling and Tracing
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
References
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Muthu, M.; Kumar, R.; Syed Khaja, A.S.; Gilthorpe, J.D.; Persson, J.L.; Nordström, A. GLUL Ablation Can Confer Drug Resistance to Cancer Cells via a Malate-Aspartate Shuttle-Mediated Mechanism. Cancers 2019, 11, 1945. https://doi.org/10.3390/cancers11121945
Muthu M, Kumar R, Syed Khaja AS, Gilthorpe JD, Persson JL, Nordström A. GLUL Ablation Can Confer Drug Resistance to Cancer Cells via a Malate-Aspartate Shuttle-Mediated Mechanism. Cancers. 2019; 11(12):1945. https://doi.org/10.3390/cancers11121945
Chicago/Turabian StyleMuthu, Magesh, Ranjeet Kumar, Azharuddin Sajid Syed Khaja, Jonathan D. Gilthorpe, Jenny L. Persson, and Anders Nordström. 2019. "GLUL Ablation Can Confer Drug Resistance to Cancer Cells via a Malate-Aspartate Shuttle-Mediated Mechanism" Cancers 11, no. 12: 1945. https://doi.org/10.3390/cancers11121945