Proteomic Characterization of a 3D HER2+ Breast Cancer Model Reveals the Role of Mitochondrial Complex I in Acquired Resistance to Trastuzumab
Abstract
:1. Introduction
2. Results
2.1. Three-Dimensional Architecture and Growth Kinetics of BT-474 Spheroids
2.2. Proteomic Characterization of BT-474 Spheroids
2.3. Identification of Differentially Expressed Proteins and Network Analysis
2.4. Functional Validations Suggest Mitochondrial Complex I Involvement
3. Discussion
4. Materials and Methods
4.1. BT-474 Tumor Spheroid Preparation and Treatment
4.1.1. Drugs and Treatments
4.1.2. Cell Cultures and Generation of Tumor Spheroids
4.1.3. Tumor Spheroid Growth
4.1.4. Tumor Spheroid Histology
4.1.5. Tumor Spheroid Viability
4.1.6. Glucose Consumption and Lactate Production Determinations
4.2. LC-MS/MS Analysis of Responder and Non-Responder BT-474 Tumor Spheroids
4.2.1. In-Solution Digestion
4.2.2. Liquid Chromatography
4.2.3. Mass Spectrometry
4.2.4. Proteomic Data Processing and Data Mining
4.2.5. Network Analysis
4.3. Analysis of Clinical Data
4.4. Statistical Analysis
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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Tapia, I.J.; Perico, D.; Wolos, V.J.; Villaverde, M.S.; Abrigo, M.; Di Silvestre, D.; Mauri, P.; De Palma, A.; Fiszman, G.L. Proteomic Characterization of a 3D HER2+ Breast Cancer Model Reveals the Role of Mitochondrial Complex I in Acquired Resistance to Trastuzumab. Int. J. Mol. Sci. 2024, 25, 7397. https://doi.org/10.3390/ijms25137397
Tapia IJ, Perico D, Wolos VJ, Villaverde MS, Abrigo M, Di Silvestre D, Mauri P, De Palma A, Fiszman GL. Proteomic Characterization of a 3D HER2+ Breast Cancer Model Reveals the Role of Mitochondrial Complex I in Acquired Resistance to Trastuzumab. International Journal of Molecular Sciences. 2024; 25(13):7397. https://doi.org/10.3390/ijms25137397
Chicago/Turabian StyleTapia, Ivana J., Davide Perico, Virginia J. Wolos, Marcela S. Villaverde, Marianela Abrigo, Dario Di Silvestre, Pierluigi Mauri, Antonella De Palma, and Gabriel L. Fiszman. 2024. "Proteomic Characterization of a 3D HER2+ Breast Cancer Model Reveals the Role of Mitochondrial Complex I in Acquired Resistance to Trastuzumab" International Journal of Molecular Sciences 25, no. 13: 7397. https://doi.org/10.3390/ijms25137397