Multifaceted Mechanisms of Cisplatin Resistance in Long-Term Treated Urothelial Carcinoma Cell Lines
Abstract
:1. Introduction
2. Results
2.1. Long-Term Cisplatin Treated (LTTs) Urothelial Carcinoma Cell Lines Recover from Cisplatin-Induced Stress
2.2. Cisplatin Exporter and Detoxifying Molecules Are Differentially Expressed in LTT Lines
2.3. DNA-Cisplatin Adduct Formation and Extent of DNA Damage Is Reduced in LTTs
2.4. LTTs Are More Resistant to Cisplatin-Induced Apoptosis
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Treatment
4.2. Measurements of Cell Viability, Clonogenicity, and Proliferation
4.3. Molecular Analyses
4.4. Measurement of Caspase 3/7 Activity
4.5. Immunofluorescence
4.6. Quantification of Pt-(GpG) Adducts
4.7. Western Blot Analysis
4.8. Flow Cytometry
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
UCC | Urothelial carcinoma cell line |
SDHA | Succinate dehydrogenase complex |
CTR1 | Solute carrier family 31 member 1 |
ATP7A | ATPase copper transporting alpha |
ATP7B | ATPase copper transporting beta |
MT | Metallothionein |
MRP2 | Multi-drug resistance protein 2 |
ERCC1 | ERCC excision repair 1, Endonuclease non-catalytic subunit |
ERCC2 | ERCC excision repair 2, TFIIH core complex helicase subunit |
MLH1 | MutL homolog 1 |
MSH2 | MutS homolog 2 |
POLH | DNA polymerase eta |
BCL-2 | B-Cell CLL/Lymphoma 2 |
BCL-XL | BCL2 like 1 |
BIRC5 | Baculoviral IAP repeat containing 5 |
γH2AX | Phosphorylated H2A histone family member X |
Pt-adduct | Platinum-adduct |
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Skowron, M.A.; Melnikova, M.; Van Roermund, J.G.H.; Romano, A.; Albers, P.; Thomale, J.; Schulz, W.A.; Niegisch, G.; Hoffmann, M.J. Multifaceted Mechanisms of Cisplatin Resistance in Long-Term Treated Urothelial Carcinoma Cell Lines. Int. J. Mol. Sci. 2018, 19, 590. https://doi.org/10.3390/ijms19020590
Skowron MA, Melnikova M, Van Roermund JGH, Romano A, Albers P, Thomale J, Schulz WA, Niegisch G, Hoffmann MJ. Multifaceted Mechanisms of Cisplatin Resistance in Long-Term Treated Urothelial Carcinoma Cell Lines. International Journal of Molecular Sciences. 2018; 19(2):590. https://doi.org/10.3390/ijms19020590
Chicago/Turabian StyleSkowron, Margaretha A., Margarita Melnikova, Joep G. H. Van Roermund, Andrea Romano, Peter Albers, Jürgen Thomale, Wolfgang A. Schulz, Günter Niegisch, and Michèle J. Hoffmann. 2018. "Multifaceted Mechanisms of Cisplatin Resistance in Long-Term Treated Urothelial Carcinoma Cell Lines" International Journal of Molecular Sciences 19, no. 2: 590. https://doi.org/10.3390/ijms19020590