A Novel Late-Stage Autophagy Inhibitor That Efficiently Targets Lysosomes Inducing Potent Cytotoxic and Sensitizing Effects in Lung Cancer
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Reagents
2.2. LAI-1 Synthesis
2.3. Transmembrane Anion Transport Experiments in Vesicles
2.4. Cell Culture
2.5. Cell Viability
2.6. Western Blotting
2.7. Flow Cytometry
2.8. Live-Cell Imaging
2.9. Immunofluorescence
2.10. Acridine Orange Staining
2.11. Intracellular pH Determination
2.12. Statistical Analysis
3. Results
3.1. LAI-1 Synthesis and Anion Transport Capacity Evaluation
3.2. LAI-1 Is Specifically Internalized to the Lysosomes
3.3. Autophagy Inhibitors Affect Lung Cancer Cell Viability
3.4. LAI-1 Is a Late-Phase Autophagy Inhibitor
3.5. LAI-1 Effectively and Rapidly Blocks Autophagy in Lung Cancer Cells
3.6. LAI-1 Blocks Autophagosomes with Lysosomes Fusion
3.7. LAI-1 Treatment Modifies Lysosomal and Cytosolic pH
3.8. Cell Death Induction after LAI-1 Treatment
3.9. LAI-1 Induces Cell Death but Not Cell Cycle Arrest
3.10. LAI-1 Sensitizes Lung Cancer Cells to Cisplatin Treatment
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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LAI-1 (µM) | A549 | SW900 | DMS53 |
---|---|---|---|
IC25 | 13.67 ± 0.82 | 13.27 ± 0.78 | 19.15 ± 2.52 |
IC50 | 15.47 ± 0.79 | 16.10 ± 0.93 | 21.81 ± 1.69 |
IC75 | 17.51 ± 0.75 | 18.56 ± 0.80 | 23.94 ± 2.03 |
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Molero-Valenzuela, A.; Fontova, P.; Alonso-Carrillo, D.; Carreira-Barral, I.; Torres, A.A.; García-Valverde, M.; Benítez-García, C.; Pérez-Tomás, R.; Quesada, R.; Soto-Cerrato, V. A Novel Late-Stage Autophagy Inhibitor That Efficiently Targets Lysosomes Inducing Potent Cytotoxic and Sensitizing Effects in Lung Cancer. Cancers 2022, 14, 3387. https://0-doi-org.brum.beds.ac.uk/10.3390/cancers14143387
Molero-Valenzuela A, Fontova P, Alonso-Carrillo D, Carreira-Barral I, Torres AA, García-Valverde M, Benítez-García C, Pérez-Tomás R, Quesada R, Soto-Cerrato V. A Novel Late-Stage Autophagy Inhibitor That Efficiently Targets Lysosomes Inducing Potent Cytotoxic and Sensitizing Effects in Lung Cancer. Cancers. 2022; 14(14):3387. https://0-doi-org.brum.beds.ac.uk/10.3390/cancers14143387
Chicago/Turabian StyleMolero-Valenzuela, Adrià, Pere Fontova, Daniel Alonso-Carrillo, Israel Carreira-Barral, Ana Aurora Torres, María García-Valverde, Cristina Benítez-García, Ricardo Pérez-Tomás, Roberto Quesada, and Vanessa Soto-Cerrato. 2022. "A Novel Late-Stage Autophagy Inhibitor That Efficiently Targets Lysosomes Inducing Potent Cytotoxic and Sensitizing Effects in Lung Cancer" Cancers 14, no. 14: 3387. https://0-doi-org.brum.beds.ac.uk/10.3390/cancers14143387