Continuous Inflammatory Stimulation Leads via Metabolic Plasticity to a Prometastatic Phenotype in Triple-Negative Breast Cancer Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Cultures and Cytokine Stimulation
2.2. Morphology Analyses and IN Cell Studies
2.3. Transcriptome Analyses
2.4. Cell Counts and XTT Assays
2.5. FACS Analyses
2.6. Spheroid Formation
2.7. Seahorse XF Analysis
2.8. Determination of Lactate and ATP Levels
2.9. ELISA Assays
2.10. RNA Extraction and Quantitative RT-PCR
2.11. Western Blot Analysis
2.12. Transwell Migration Assay
2.13. In Vivo Plug Assay
2.14. Statistical Analyses
3. Results
3.1. Continuous Stimulation by Proinflammatory Cytokines Induces Morphological Alterations in TNBC Cells
3.2. Continuous Stimulation by Proinflammatory Cytokines Modifies Gene Expression in TNBC Cells
3.3. Continuous Stimulation by Proinflammatory Cytokines Promotes TNBC Cell Dispersion but Does Not Have a Consistent Impact on Stemness, Chemoresistance and Tumor Cell Proliferation
3.4. Continuous Stimulation by Proinflammatory Cytokines Leads to Increased Glycolysis and OXPHOS in TNBC Cells
3.5. Continuous Stimulation by Proinflammatory Cytokines Leads via Metabolic Transition to Increased Activation of p65 in TNBC Cells
3.6. Continuous Stimulation of TNBC Cells by Proinflammatory Cytokines Leads in a Glycolysis-Dependent Manner to Increased Release of Proinflammatory Mediators and to Monocyte and Neutrophil Migration In Vivo
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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Morein, D.; Rubinstein-Achiasaf, L.; Brayer, H.; Dorot, O.; Pichinuk, E.; Ben-Yaakov, H.; Meshel, T.; Pasmanik-Chor, M.; Ben-Baruch, A. Continuous Inflammatory Stimulation Leads via Metabolic Plasticity to a Prometastatic Phenotype in Triple-Negative Breast Cancer Cells. Cells 2021, 10, 1356. https://doi.org/10.3390/cells10061356
Morein D, Rubinstein-Achiasaf L, Brayer H, Dorot O, Pichinuk E, Ben-Yaakov H, Meshel T, Pasmanik-Chor M, Ben-Baruch A. Continuous Inflammatory Stimulation Leads via Metabolic Plasticity to a Prometastatic Phenotype in Triple-Negative Breast Cancer Cells. Cells. 2021; 10(6):1356. https://doi.org/10.3390/cells10061356
Chicago/Turabian StyleMorein, Dina, Linor Rubinstein-Achiasaf, Hadar Brayer, Orly Dorot, Edward Pichinuk, Hagar Ben-Yaakov, Tsipi Meshel, Metsada Pasmanik-Chor, and Adit Ben-Baruch. 2021. "Continuous Inflammatory Stimulation Leads via Metabolic Plasticity to a Prometastatic Phenotype in Triple-Negative Breast Cancer Cells" Cells 10, no. 6: 1356. https://doi.org/10.3390/cells10061356