Impairment of Nucleolin Activity and Phosphorylation by a Trachylobane Diterpene from Psiadia punctulata in Cancer Cells
Abstract
:1. Introduction
2. Results and Discussion
2.1. CETSA-Based Screening of Putative hNcl Interactors
2.2. Validation of the Interaction of Compound 12 with hNcl
2.3. Molecular Modeling of the 12/hNcl Complex
2.4. Modulation of hNcl Activity by Compound 12
2.5. Biological Effects of Compound 12 Treatment on Cancer Cells
3. Materials and Methods
3.1. Materials
3.2. Cell Culture and Treatment
3.3. MTT Assay
3.4. CETSA-Based Screening
3.5. Determination of the Apparent Melting Temperature (Tm) of hNcl in HeLa Cells
3.6. Determination of the EC50
3.7. Drug Affinity Responsive Target Stability (DARTS) Experiments
3.8. Surface Plasmon Resonance
3.9. Docking Studies
3.10. Cytosol, Membrane, and Nuclear Extracts
3.11. RNA Isolation and Quantitative Real-Time-PCR (qRT-PCR)
3.12. Cell Cycle and Apoptosis Analysis
3.13. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Bellone, M.L.; Fiengo, L.; Cerchia, C.; Cotugno, R.; Bader, A.; Lavecchia, A.; De Tommasi, N.; Piaz, F.D. Impairment of Nucleolin Activity and Phosphorylation by a Trachylobane Diterpene from Psiadia punctulata in Cancer Cells. Int. J. Mol. Sci. 2022, 23, 11390. https://doi.org/10.3390/ijms231911390
Bellone ML, Fiengo L, Cerchia C, Cotugno R, Bader A, Lavecchia A, De Tommasi N, Piaz FD. Impairment of Nucleolin Activity and Phosphorylation by a Trachylobane Diterpene from Psiadia punctulata in Cancer Cells. International Journal of Molecular Sciences. 2022; 23(19):11390. https://doi.org/10.3390/ijms231911390
Chicago/Turabian StyleBellone, Maria Laura, Lorenzo Fiengo, Carmen Cerchia, Roberta Cotugno, Ammar Bader, Antonio Lavecchia, Nunziatina De Tommasi, and Fabrizio Dal Piaz. 2022. "Impairment of Nucleolin Activity and Phosphorylation by a Trachylobane Diterpene from Psiadia punctulata in Cancer Cells" International Journal of Molecular Sciences 23, no. 19: 11390. https://doi.org/10.3390/ijms231911390