Checkpoint kinase inhibitor AZD7762 strongly sensitises urothelial carcinoma cells to gemcitabine.
Checkpoint kinase inhibitor AZD7762 strongly sensitises urothelial carcinoma cells to gemcitabine.
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DOI：
10.1186/s13046-016-0473-1
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发表时间：
2017-01-03
期刊：
影响因子：
--
通讯作者：
Schulz WA
中科院分区：
文献类型：
--
作者：
Isono M;Hoffmann MJ;Pinkerneil M;Sato A;Michaelis M;Cinatl J Jr;Niegisch G;Schulz WA
膀胱癌是世界范围内发病率和死亡率的主要原因，迫切需要更有效的化疗。因此，我们探索了吉西他滨和检查点激酶1/2 （CHK1/2）抑制剂AZD7762联合治疗膀胱癌的疗效。用吉西他滨和AZD7762治疗尿路上皮癌细胞系和各种非恶性尿路上皮细胞，评估细胞活力、克隆原性、细胞周期分布和凋亡。用γ - h2a评价DNA损伤。X和53-BP1染色和检查点激活后进行Western blotting。将CHK1和CHK2的药理抑制与使用sirna下调CHK1或CHK2进行比较。吉西他滨和AZD7762联合使用，与单独治疗相比，可协同降低尿路上皮癌细胞活力和集落形成。非恶性尿路上皮细胞对这种药物组合的敏感性较低。吉西他滨联合AZD7762抑制细胞周期进程，导致细胞在s期积累。此外，该组合诱导了明显的凋亡水平，亚g1细胞的比例增加，裂解的PARP水平增加，caspase 3/7活性增加。机制研究表明，AZD7762通过干扰CHK1抑制了吉西他滨诱导的双链断裂的修复，这是因为sirna介导的CHK1而非CHK2的缺失模仿了AZD7762的作用。AZD7762通过抑制DNA修复和干扰检查点增强尿路上皮癌细胞对吉西他滨的敏感性。吉西他滨联合CHK1抑制有望治疗尿路上皮癌。本文的在线版本（doi:10.1186/s13046-016-0473-1）包含补充材料，仅供授权用户使用。
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. 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. 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. 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. The online version of this article (doi:10.1186/s13046-016-0473-1) contains supplementary material, which is available to authorized users.
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