Upregulation of p75NTR by Histone Deacetylase Inhibitors Sensitizes Human Neuroblastoma Cells to Targeted Immunotoxin-Induced Apoptosis
Abstract
:1. Introduction
2. Results
2.1. Upregulation of p75NTR by Valproic Acid and Entinostat in Different Human Neuroblastoma Cell Lines
2.2. Enhancement of p75NTR Cell Surface Expression by VPA and Entinostat
2.3. Internalization of p75NTR and Intracellular Localization of Saporin-S6 following Exposure to p75IgG-Sap
2.4. Entinostat Predisposes Neuroblastoma Cells to p75IgG-Sap Cytotoxicity
2.5. Exposure to p75IgG-Sap Potentiates Entinostat-Induced Apoptosis of Neuroblastoma Cells
2.6. Entinostat Upregulates p75NTR and Induces Apoptosis in Neuroblastoma Multicell Spheroids
2.7. Treatment with p75IgG-Sap Enhances the Cytotoxic Effect of Entinostat in Neuroblastoma Multicell Spheroids
2.8. Entinostat Upregulates p75NTR Expression in Neuroblastoma Xenografts
2.9. Antineuroblastoma Effect of Intratumoral Administration of p75IgG-Sap
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Cell Culture
4.3. Cell Treatment and Cell Lysate Preparation
4.4. Biotinylation of Cell Surface Proteins
4.5. Western Blot Analysis
4.6. Immunofluorescence Analysis
4.7. Analysis of Living Cell Morphology
4.8. Assay of Cell Viability
4.9. Assay of Caspase Activity
4.10. Terminal Transferase dUTP Nick End-Labeling Assay (TUNEL Assay)
4.11. Tumour Spheroid Generation and Analysis
4.12. Neuroblastoma Cell Xenografts
4.13. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Dedoni, S.; Olianas, A.; Manconi, B.; Collu, M.; Tuveri, B.; Vincis, M.E.; Olianas, M.C.; Onali, P. Upregulation of p75NTR by Histone Deacetylase Inhibitors Sensitizes Human Neuroblastoma Cells to Targeted Immunotoxin-Induced Apoptosis. Int. J. Mol. Sci. 2022, 23, 3849. https://doi.org/10.3390/ijms23073849
Dedoni S, Olianas A, Manconi B, Collu M, Tuveri B, Vincis ME, Olianas MC, Onali P. Upregulation of p75NTR by Histone Deacetylase Inhibitors Sensitizes Human Neuroblastoma Cells to Targeted Immunotoxin-Induced Apoptosis. International Journal of Molecular Sciences. 2022; 23(7):3849. https://doi.org/10.3390/ijms23073849
Chicago/Turabian StyleDedoni, Simona, Alessandra Olianas, Barbara Manconi, Maria Collu, Barbara Tuveri, Maria Elena Vincis, Maria C. Olianas, and Pierluigi Onali. 2022. "Upregulation of p75NTR by Histone Deacetylase Inhibitors Sensitizes Human Neuroblastoma Cells to Targeted Immunotoxin-Induced Apoptosis" International Journal of Molecular Sciences 23, no. 7: 3849. https://doi.org/10.3390/ijms23073849