Dasatinib Treatment Increases Sensitivity to c-Met Inhibition in Triple-Negative Breast Cancer Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Lines and Reagents
2.2. Proliferation Assays
2.3. Doubling Time Assays
2.4. Short-Term Resistance Assay
2.5. Invasion/Migration Assays
2.6. Dasatinib Accumulation Assays
2.7. Protein Extraction and Western Blotting
2.8. Luminex Magnetic Bead Assays
2.9. DNA Extraction and Nested PCR Amplification of Src Exons 9–12
2.10. Cycle Sequencing of PCR Products
2.11. Statistical Analysis
3. Results
3.1. Dasatinib Exposure Induces a Resistant Phenotype
3.2. Phosphorylation of Src Is Altered in Dasatinib-Resistant Cells
3.3. cMet Signalling Is Increased in Dasatinib-Resistant Cells
3.4. cMET Inhibition Blocks Dasatinib Resistance
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Cell Line | Control | D 50 nM | D 100 nM |
---|---|---|---|
MDA-MB-231 | 17.6 ± 1.2 | 32.2 ± 3.3* | 46.8 ± 5.1* |
231 DasB | 19.1 ± 2.4 | 21.0 ± 0.2 | 21.2 ± 3.1 |
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Gaule, P.; Mukherjee, N.; Corkery, B.; Eustace, A.J.; Gately, K.; Roche, S.; O’Connor, R.; O’Byrne, K.J.; Walsh, N.; Duffy, M.J.; et al. Dasatinib Treatment Increases Sensitivity to c-Met Inhibition in Triple-Negative Breast Cancer Cells. Cancers 2019, 11, 548. https://doi.org/10.3390/cancers11040548
Gaule P, Mukherjee N, Corkery B, Eustace AJ, Gately K, Roche S, O’Connor R, O’Byrne KJ, Walsh N, Duffy MJ, et al. Dasatinib Treatment Increases Sensitivity to c-Met Inhibition in Triple-Negative Breast Cancer Cells. Cancers. 2019; 11(4):548. https://doi.org/10.3390/cancers11040548
Chicago/Turabian StyleGaule, Patricia, Nupur Mukherjee, Brendan Corkery, Alex J. Eustace, Kathy Gately, Sandra Roche, Robert O’Connor, Kenneth J. O’Byrne, Naomi Walsh, Michael J. Duffy, and et al. 2019. "Dasatinib Treatment Increases Sensitivity to c-Met Inhibition in Triple-Negative Breast Cancer Cells" Cancers 11, no. 4: 548. https://doi.org/10.3390/cancers11040548