p53-Mediated Radiosensitization of 177Lu-DOTATATE in Neuroblastoma Tumor Spheroids
Abstract
:1. Introduction
2. Materials and Methods
2.1. Statistical Analysis
2.2. Cell Lines
2.3. Drug and Radioconjugate Preparation
2.4. XTT Viability Assay
2.5. Sequencing of the p53 Gene
2.6. Validation of VIP116 Specificity
2.7. Cellular Specificty of 177Lu-DOTATATE and Ligandtracer Experiments
2.8. Multicellular Tumor Spheroids
2.9. Cell Lysate Preparation and Western Blot
2.10. Flow Cytometry
3. Results
3.1. Labeling
3.2. Characterization of Cell Lines
3.3. 177Lu-DOTATATE and VIP116 Monotherapy Inhibit Spheroid Growth
3.4. Combination Therapy Induces Varying Degree of Synergy in IMR-32 and NB2 Spheroids
3.5. Combination Treatment Activates p53 Pathway with Differential Effects on MYCN and MDM2
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Cell Line | p53 Status | MYCN Status | XTT VIP116 (μM) | Sph. VIP116 (μM) | Sph. 177Lu-DOT (kBq/well) | Ref. |
---|---|---|---|---|---|---|
IMR-32 | wt | amp | 13.2 (11.1–15.4) | 15.3 (12.7–18.1) | 0.12 (0.10–0.14) | [35,47,48] |
NB1 | wt | amp | 2.9 (2.3–3.7) | 0.9 (0.8–1.0) | 1.1 (0.8–1.5) | [39] |
NB2 | wt | amp | 2.8 (2.6–3.0) | 1.4 (1.2–1.6) | 0.88 (0.67–1.21) | [38] |
N2a | mut 1 | amp | 238.2 (60.5–105) 3 | n.e. | n.e. | [49] |
SKNAS | del 2 | n.amp | 40.8 (27.8–45.4) 3 | n.e. | n.e. | [35,50,51] |
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Lundsten, S.; Berglund, H.; Jha, P.; Krona, C.; Hariri, M.; Nelander, S.; Lane, D.P.; Nestor, M. p53-Mediated Radiosensitization of 177Lu-DOTATATE in Neuroblastoma Tumor Spheroids. Biomolecules 2021, 11, 1695. https://doi.org/10.3390/biom11111695
Lundsten S, Berglund H, Jha P, Krona C, Hariri M, Nelander S, Lane DP, Nestor M. p53-Mediated Radiosensitization of 177Lu-DOTATATE in Neuroblastoma Tumor Spheroids. Biomolecules. 2021; 11(11):1695. https://doi.org/10.3390/biom11111695
Chicago/Turabian StyleLundsten, Sara, Hanna Berglund, Preeti Jha, Cecilia Krona, Mehran Hariri, Sven Nelander, David P. Lane, and Marika Nestor. 2021. "p53-Mediated Radiosensitization of 177Lu-DOTATATE in Neuroblastoma Tumor Spheroids" Biomolecules 11, no. 11: 1695. https://doi.org/10.3390/biom11111695