SPARC Inhibits Metabolic Plasticity in Ovarian Cancer
Abstract
:1. Introduction
2. Results
2.1. Loss of Stromal-SPARC Is Associated with Enhanced Glycolysis in Syngeneic ID8 Intraperitoneal Tumors
2.2. Effect of Loss of Stromal-SPARC on TCA Cycle and Pentose Phosphate Pathway (PPP)
2.3. Loss of Stromal-Sparc Increases OXPHOS with Overexpression of Mitochondrial ETC Enzymes
2.4. Effect of SPARC on Mitochondrial Functions
2.5. Perturbed Redox Homeostasis in Tumors Dissected from SP−/− Mice
2.6. Targeting Mitochondrial ETC by Phenformin Mitigates the Accelerated Growth of OvCa Syngeneic Tumors
3. Discussion
4. Material and Methods
4.1. Cell Culture and Reagents
4.2. In Vivo Syngeneic Model
4.3. Mouse Therapeutic Trial with Phenformin
4.4. Immunohistochemistry
4.5. Laser Captured Microdissection (LCM)
4.6. RNA Extraction, Amplification, Labeling and Analysis
4.7. Quantitative RT-PCR Analysis
4.8. Metabolomic Profiling Sample Preparation
4.9. Data Extraction, Compound Identification, Metabolite Quantification and Data Normalization
4.10. Intracellular ROS Formation in Tumor Tissues
4.11. Total ATP Measurement, Hexokinase 2 and TPI1 Enzyme Activity Assays
4.12. Transmission Electron Microscopy (TEM)
4.13. Glycolysis and Mitochondrial Stress Assays
4.14. Measurement of Mitochondrial Contents
4.15. Statistical Analysis
4.16. Bioinformatics Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Naczki, C.; John, B.; Patel, C.; Lafferty, A.; Ghoneum, A.; Afify, H.; White, M.; Davis, A.; Jin, G.; Kridel, S.; et al. SPARC Inhibits Metabolic Plasticity in Ovarian Cancer. Cancers 2018, 10, 385. https://doi.org/10.3390/cancers10100385
Naczki C, John B, Patel C, Lafferty A, Ghoneum A, Afify H, White M, Davis A, Jin G, Kridel S, et al. SPARC Inhibits Metabolic Plasticity in Ovarian Cancer. Cancers. 2018; 10(10):385. https://doi.org/10.3390/cancers10100385
Chicago/Turabian StyleNaczki, Christine, Bincy John, Chirayu Patel, Ashlyn Lafferty, Alia Ghoneum, Hesham Afify, Michael White, Amanda Davis, Guangxu Jin, Steven Kridel, and et al. 2018. "SPARC Inhibits Metabolic Plasticity in Ovarian Cancer" Cancers 10, no. 10: 385. https://doi.org/10.3390/cancers10100385