HSP90抑制剂Ganetespib增强肺癌细胞对顺铂的敏感性
| 作者: |
1姜桔红,
1顾莹莹,
1刘静,
1廖炫之,
1赵瑾,
1李龙光,
1付琳,
1李谨
1 广州医科大学附属第一医院，呼吸疾病国家重点实验室，广州呼吸疾病研究所，广州 510182 |
| 通讯: |
李谨
Email: lij1250@hotmail.com |
| DOI: | 10.3978/j.issn.2095-6959.2017.07.004 |
| 基金: | 广州市属高校科研项目, 1201620051 国家自然科学基金, 81272901 |
摘要
目的：研究HSP90抑制剂ganetespib对DNA修复相关蛋白表达的下调作用及其对顺铂(cis-diamminedichloroplatinum，DDP)的增敏作用。方法：采用Western印迹法检测ganetespib对肺癌细胞A549和H1975的DNA修复蛋白BRCA1及RAD51表达的下调作用；免疫荧光法检测细胞RAD51及核磷酸化的H2AX组蛋白(γ-H2AX)核焦点形成；流式细胞技术检测细胞凋亡；CKK-8试剂盒检测细胞的增殖，中效原理分析判断两种药物联合应用的协同作用；并通过SCID皮下移植瘤模型检测ganetespib与DDP联合应用的体内抑瘤作用。结果：Ganetespid明显下调DNA修复蛋白BRCA1和RAD51的表达，ganetespid及DDP处理诱导DNA重组修复标记RAD51核焦点的形成率为13%，明显少于单一DDP处理组(59%)。Ganetespid和DDP联合应用的细胞凋亡率为23%，明显高于单一ganetespid组(15%)或单一DDP组(16%)，差异有统计学意义(P<0.01)。SCID小鼠移植瘤模型中ganetespid与DDP联合应用对肿瘤生长抑制作用明显优于单一ganetespib或单一DDP处理组，差异有统计学意义(P<0.01)。结论：ganetespib可下调肺癌细胞DNA修复蛋白的表达并增加对DDP的敏感性。
关键词：
肺癌细胞
HSP90抑制剂
顺铂
DNA修复
HSP90 inhibitor ganetespib enhances the sensitivity of lung cancer cell to cis-diamminedichloroplatinum
CorrespondingAuthor: LI Jin Email: lij1250@hotmail.com
DOI: 10.3978/j.issn.2095-6959.2017.07.004
Abstract
Objective: To investigate the potential effects of HSP90 inhibitor ganetespib in the sensitization of cis-diamminedichloroplatinum (DDP) through down regulation of DNA repair protein BRCA1 and RAD51. Methods: Many aspects of the inhibition were studied. Western blot was used to detect the expression of BRCA1 and RAD51 in A549 and H1975 cell. Intracellularly, the formation of γ-H2AX and RAD51 foci were detected by immunofluorescence staining. Apoptotic cells were analyzed by Flow cytometry. Cell proliferation was examined by using cell counting kit-8 (CCK-8), and the combined drug interaction was analyzed using the median-effect method. Lastly, in vivo anti-tumor activity was evaluated in A549 xenograft SCID mice. Results: The expressions of BRCA1 and RAD51 were significantly down regulated by ganetespib treatment. Combined treatment of ganetespib and DDP induced significantly less RAD51 foci than DDP treatment alone and also induced more apoptosis than DDP treatment alone. The combination of ganetespib and DDP in A549 xenograft model resulted in a superior therapeutic response compared to treatment with just single-agent DDP. Conclusion: Ganetespib demonstrated the ability to down regulate the expression of DNA repair proteins in lung cancer cell and enhance the sensitivity to DDP.