PARP inhibitor veliparib and HDAC inhibitor SAHA synergistically co-target the UHRF1/BRCA1 DNA damage repair complex in prostate cancer cells.
PARP inhibitor veliparib and HDAC inhibitor SAHA synergistically co-target the UHRF1/BRCA1 DNA damage repair complex in prostate cancer cells.
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PARP 抑制剂 veliparib 和 HDAC 抑制剂 SAHA 协同靶向前列腺癌细胞中的 UHRF1/BRCA1 DNA 损伤修复复合物。
DOI：
10.1186/s13046-018-0810-7
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发表时间：
2018-07-16
期刊：
影响因子：
--
通讯作者：
Li X
中科院分区：
文献类型：
--
作者：
Yin L;Liu Y;Peng Y;Peng Y;Yu X;Gao Y;Yuan B;Zhu Q;Cao T;He L;Gong Z;Sun L;Fan X;Li X
聚ADP核糖聚合酶（PARP）抑制剂奥拉帕尼已被批准用于治疗BRCA突变的前列腺癌（PCa），另一种PARP抑制剂veliparib正在临床试验中进行测试。然而，veliparib仅显示出中度抗癌作用，PCa患者需要联合治疗。组蛋白去乙酰化酶（HDAC）抑制剂已被测试以改善PARP抑制剂对PCa细胞的抗癌功效，但确切的机制仍然难以捉摸。用维利帕尼或SAHA单独或组合处理几种类型的PCa细胞和前列腺上皮细胞系RWPE-1。采用紫外线结晶法检测细胞活力或克隆形成能力，Annexin V-FITC/PI染色和流式细胞仪检测细胞凋亡，Western blot检测PARP裂解，γ H2 AX抗体染色检测细胞DNA损伤，荧光显微镜观察DNA损伤灶，Western blot检测γ H2 AX水平; Western blot和细胞免疫荧光染色检测UHRF 1和BRCA 1蛋白水平，免疫共沉淀法检测药物作用后UHRF 1和BRCA 1蛋白的相互作用。在DU 145异种移植模型中验证了联合治疗的抗肿瘤作用。PCa细胞对维利帕尼或SAHA显示出不同的敏感性。两种药物联合给药可协同降低细胞活力和克隆形成，并协同诱导细胞凋亡和DNA损伤，同时对正常前列腺上皮细胞没有可检测到的毒性。在机制上，单独的维利帕尼或SAHA降低了BRCA 1或UHRF 1蛋白水平，维利帕尼和SAHA的共同治疗通过靶向UHRF 1/BRCA 1蛋白复合物协同降低了BRCA 1蛋白水平，UHRF 1的消耗导致BRCA 1蛋白降解，而UHRF 1的升高损害了共同治疗降低的BRCA 1蛋白水平。两种药物的联合给药协同降低异种移植物的生长。我们的研究表明，HDAC和PARP抑制剂的协同致死性是由于促进DNA损伤和抑制HR DNA损伤修复途径，特别是靶向UHRF 1/BRCA 1蛋白复合物。维利帕尼和SAHA的协同致死性显示了未来PCa临床试验的巨大潜力。本文的在线版本（10.1186/s13046-018-0810-7）包含补充材料，可供授权用户使用。
The poly ADP ribose polymerase (PARP) inhibitor olaparib has been approved for treating prostate cancer (PCa) with BRCA mutations, and veliparib, another PARP inhibitor, is being tested in clinical trials. However, veliparib only showed a moderate anticancer effect, and combination therapy is required for PCa patients. Histone deacetylase (HDAC) inhibitors have been tested to improve the anticancer efficacy of PARP inhibitors for PCa cells, but the exact mechanisms are still elusive. Several types of PCa cells and prostate epithelial cell line RWPE-1 were treated with veliparib or SAHA alone or in combination. Cell viability or clonogenicity was tested with violet crystal assay; cell apoptosis was detected with Annexin V-FITC/PI staining and flow cytometry, and the cleaved PARP was tested with western blot; DNA damage was evaluated by staining the cells with γH2AX antibody, and the DNA damage foci were observed with a fluorescent microscopy, and the level of γH2AX was tested with western blot; the protein levels of UHRF1 and BRCA1 were measured with western blot or cell immunofluorescent staining, and the interaction of UHRF1 and BRCA1 proteins was detected with co-immunoprecipitation when cells were treated with drugs. The antitumor effect of combinational therapy was validated in DU145 xenograft models. PCa cells showed different sensitivity to veliparib or SAHA. Co-administration of both drugs synergistically decreased cell viability and clonogenicity, and synergistically induced cell apoptosis and DNA damage, while had no detectable toxicity to normal prostate epithelial cells. Mechanistically, veliparib or SAHA alone reduced BRCA1 or UHRF1 protein levels, co-treatment with veliparib and SAHA synergistically reduced BRCA1 protein levels by targeting the UHRF1/BRCA1 protein complex, the depletion of UHRF1 resulted in the degradation of BRCA1 protein, while the elevation of UHRF1 impaired co-treatment-reduced BRCA1 protein levels. Co-administration of both drugs synergistically decreased the growth of xenografts. Our studies revealed that the synergistic lethality of HDAC and PARP inhibitors resulted from promoting DNA damage and inhibiting HR DNA damage repair pathways, in particular targeting the UHRF1/BRCA1 protein complex. The synergistic lethality of veliparib and SAHA shows great potential for future PCa clinical trials. The online version of this article (10.1186/s13046-018-0810-7) contains supplementary material, which is available to authorized users.
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