Checkpoint kinase inhibitor AZD7762 strongly sensitises urothelial carcinoma cells to gemcitabine
- Background: More effective chemotherapies are urgently needed for bladder cancer, a major cause of morbidity and mortality worldwide. We therefore explored the efficacy of the combination of gemcitabine and AZD7762, a checkpoint kinase 1/2 (CHK1/2) inhibitor, for bladder cancer. Methods: Viability, clonogenicity, cell cycle distribution and apoptosis were assessed in urothelial cancer cell lines and various non-malignant urothelial cells treated with gemcitabine and AZD7762. DNA damage was assessed by γH2A.X and 53-BP1 staining and checkpoint activation was followed by Western blotting. Pharmacological inhibition of CHK1 and CHK2 was compared to downregulation of either CHK1 or CHK2 using siRNAs. Results: Combined use of gemcitabine and AZD7762 synergistically reduced urothelial carcinoma cell viability and colony formation relative to either single treatment. Non-malignant urothelial cells were substantially less sensitive to this drug combination. Gemcitabine plus AZD7762 inhibited cell cycle progression causing cell accumulation in S-phase. Moreover, the combination induced pronounced levels of apoptosis as indicated by an increase in the fraction of sub-G1 cells, in the levels of cleaved PARP, and in caspase 3/7 activity. Mechanistic investigations showed that AZD7762 treatment inhibited the repair of gemcitabine-induced double strand breaks by interference with CHK1, since siRNA-mediated depletion of CHK1 but not of CHK2 mimicked the effects of AZD7762. Conclusions: AZD7762 enhanced sensitivity of urothelial carcinoma cells to gemcitabine by inhibiting DNA repair and disturbing checkpoints. Combining gemcitabine with CHK1 inhibition holds promise for urothelial cancer therapy.
Author: | Makoto Isono, Michèle Janine Hoffmann, Maria Pinkerneil, Akinori Sato, Martin MichaelisORCiDGND, Jindrich CinatlORCiDGND, Günter Niegisch, Wolfgang Arthur Schulz |
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URN: | urn:nbn:de:hebis:30:3-455557 |
DOI: | https://doi.org/10.1186/s13046-016-0473-1 |
ISSN: | 1756-9966 |
ISSN: | 0392-9078 |
Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/28049532 |
Parent Title (German): | Journal of experimental & clinical cancer research |
Publisher: | Springer ; BioMed Central |
Place of publication: | Berlin ; Heidelberg ; London |
Document Type: | Article |
Language: | English |
Year of Completion: | 2017 |
Date of first Publication: | 2017/01/03 |
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
Release Date: | 2018/01/30 |
Tag: | AZD7762; Bladder cancer; Checkpoint kinase; Gemcitabine; Urothelial carcinoma |
Volume: | 36 |
Issue: | 1, Art. 1 |
Page Number: | 12 |
First Page: | 1 |
Last Page: | 12 |
Note: | © The Author(s). 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
HeBIS-PPN: | 427970547 |
Institutes: | Medizin / Medizin |
Dewey Decimal Classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
Sammlungen: | Universitätspublikationen |
Licence (German): | Creative Commons - Namensnennung 4.0 |