Synergism Through WEE1 and CHK1 Inhibition in Acute Lymphoblastic Leukemia
Abstract
:1. Introduction
2. Methods
2.1. Drugs and Cell Lines
2.2. Viability and Combination Index Analysis
2.3. Cell Cycle and Apoptosis Analysis
2.4. BrdU Incorporation Assay
2.5. Clonogenic Assay and Morphological Analysis
2.6. Quantitative PCR of CHK1, CHK2 and WEE1 in Primary B-ALL Samples
2.7. Immunoblotting
2.8. Gene Expression Profiling
2.9. Statistical Analysis
3. Results
3.1. The Simultaneous Inhibition of CHK1, CHK2 and WEE1 Impairs ALL Cell Lines Viability and Triggers Apoptosis
3.2. The Inhibition of CHK1, CHK2 and WEE1 Kinases Affect Early S-phase Regulation Cells
3.3. The Combination AZD-1775 and PF-00477736 Triggers the S Phase Checkpoint and Induce DNA Damage
3.4. The Effect of the Combination Is Independent the Activity of p53 in the Regulation of Cell Cycle
3.5. CHK1 and WEE1 Are Co-Expressed in ALL Primary Cells
3.6. Inhibition of CHK1/2 and WEE1 Affect the Clonogenic Capacity of Primary ALL Cells
4. Discussion
4.1. The Inhibition of CHK1/WEE1 Kinases Highlights Potential Therapeutic Strategies Affecting S Phase
4.2. The Efficacy of the Combination Strategy Is Independent to p53 Functionality in ALL Cell Lines
4.3. Limiting the Toxicity of CHK1/WEE1 Inhibition Using Combination Strategies
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
ALL | acute lymphoblastic leukemia |
BM | bone marrow |
B-ALL | B-cell precursor acute lymphoblastic leukemia |
DDR | DNA damage response |
PB | peripheral blood |
PI | Propidium Iodide |
SD | standard deviation |
SER | serine |
T-ALL | T-cell precursor acute lymphoblastic leukemia |
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Ghelli Luserna Di Rorà, A.; Bocconcelli, M.; Ferrari, A.; Terragna, C.; Bruno, S.; Imbrogno, E.; Beeharry, N.; Robustelli, V.; Ghetti, M.; Napolitano, R.; et al. Synergism Through WEE1 and CHK1 Inhibition in Acute Lymphoblastic Leukemia. Cancers 2019, 11, 1654. https://doi.org/10.3390/cancers11111654
Ghelli Luserna Di Rorà A, Bocconcelli M, Ferrari A, Terragna C, Bruno S, Imbrogno E, Beeharry N, Robustelli V, Ghetti M, Napolitano R, et al. Synergism Through WEE1 and CHK1 Inhibition in Acute Lymphoblastic Leukemia. Cancers. 2019; 11(11):1654. https://doi.org/10.3390/cancers11111654
Chicago/Turabian StyleGhelli Luserna Di Rorà, Andrea, Matteo Bocconcelli, Anna Ferrari, Carolina Terragna, Samantha Bruno, Enrica Imbrogno, Neil Beeharry, Valentina Robustelli, Martina Ghetti, Roberta Napolitano, and et al. 2019. "Synergism Through WEE1 and CHK1 Inhibition in Acute Lymphoblastic Leukemia" Cancers 11, no. 11: 1654. https://doi.org/10.3390/cancers11111654