DKC1 Overexpression Induces a More Aggressive Cellular Behavior and Increases Intrinsic Ribosomal Activity in Immortalized Mammary Gland Cells
Abstract
:Simple Summary
Abstract
1. Introduction
2. Results
2.1. DKC1 Overexpression Is Related to Worse Prognosis for Patients, and Confers a More Aggressive Neoplastic Features to Immortalized Mammary Gland Epithelial Cells In Vitro
2.2. DKC1 Overexpression Induced a Significant Increase in Telomerase RNA Component without Affecting Telomerase Activity
2.3. DKC1 Overexpression Did Not Affect Global Pseudouridylation on rRNA, but Induced a Remodulation in snoRNAs Expression Levels
2.4. DKC1 Overexpression Increases Translation Efficiency of Ribosomes in MCF10A Cells
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Generation of DKC1 Overexpression Models
4.2. RNA Extraction and Real-Time RT-PCR
4.3. Whole Cell Protein Extraction and Western Blot Analysis
4.4. Cell Invasive, Clonogenic, and Stemness Potentials Assays
4.5. Telomerase Activity Assay
4.6. Ribosome Purification
4.7. In Vitro Translation Assays
4.8. Global Pseudouridylation Quantification
4.9. SnoRNAs Expression Array
4.10. SILNAS LC/MS Based Quantitation of Ψs
4.11. Patients’ Material
4.12. Statistical Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Guerrieri, A.N.; Zacchini, F.; Onofrillo, C.; Di Viggiano, S.; Penzo, M.; Ansuini, A.; Gandin, I.; Nobe, Y.; Taoka, M.; Isobe, T.; et al. DKC1 Overexpression Induces a More Aggressive Cellular Behavior and Increases Intrinsic Ribosomal Activity in Immortalized Mammary Gland Cells. Cancers 2020, 12, 3512. https://doi.org/10.3390/cancers12123512
Guerrieri AN, Zacchini F, Onofrillo C, Di Viggiano S, Penzo M, Ansuini A, Gandin I, Nobe Y, Taoka M, Isobe T, et al. DKC1 Overexpression Induces a More Aggressive Cellular Behavior and Increases Intrinsic Ribosomal Activity in Immortalized Mammary Gland Cells. Cancers. 2020; 12(12):3512. https://doi.org/10.3390/cancers12123512
Chicago/Turabian StyleGuerrieri, Ania Naila, Federico Zacchini, Carmine Onofrillo, Sara Di Viggiano, Marianna Penzo, Alessio Ansuini, Ilaria Gandin, Yuko Nobe, Masato Taoka, Toshiaki Isobe, and et al. 2020. "DKC1 Overexpression Induces a More Aggressive Cellular Behavior and Increases Intrinsic Ribosomal Activity in Immortalized Mammary Gland Cells" Cancers 12, no. 12: 3512. https://doi.org/10.3390/cancers12123512