摘要
目的筛选靶向PLK1 PBD的抗肿瘤小分子抑制剂,对其抗肿瘤效果进行体外评价,为抗肿瘤药物提供先导化合物。方法利用荧光偏振模型初筛PLK1 PBD抑制剂;通过MTT法寻找初筛阳性化合物的敏感细胞系;利用流式细胞仪检测化合物对细胞周期和凋亡的影响;通过分子对接技术探讨化合物与PLK1 PBD结合方式;利用划痕实验测定化合物对肿瘤细胞迁移的影响。结果从20 000个化合物中筛选得到活性化合物1097C11,MTT结果显示其能抑制多种肿瘤细胞系的增殖;流式细胞仪检测其能够促进肿瘤细胞凋亡,在25μmol/L时,晚期凋亡达到77.02%;能导致细胞G1/M期阻滞,分子对接结果显示1097C11与PLK1 PBD结构域具有良好的亲和性,细胞迁移结果显示1097C11能够抑制HCT-116细胞的迁移,在20μmol/L时,迁移率低至33.4%。结论化合物1097C11具有较好的抗肿瘤活性,并有望成为靶向PLK1 PBD的抗肿瘤先导化合物。
Objective In order to find lead compound which is capable, at least in vitro, of inhibiting tumor cells＇ growth by restraining PLK1 PBD＇s activity. Methods We used fluorescence polarization model to screen positive compounds targeting PLK1 PBD. MTT assay was utilized to find the most sensitive cell lines of 1097C11. Effects of the positive compound on cell cycle and apoptosis were all tested by flow cytometry（FCM）. The combination of compound with PLK1 PBD was determined by molecular docking. Compound＇s effect on cell movement was determined by cell migration experiment. Results The hit 1097C11 was found from 20 000 compounds. It could suppress the proliferation of HCT-116 cells. 1097C11 induced apoptosis, with apoptosis ratio reaching as high as 77.02% at 25 μmol/L concentration, and also led to G1/M phase arrest. Molecular docking showed that 1097C11 had good affinity with the PBD1 domain. Cell migration results showed that 1097C11 inhibited the migration of HCT-116 cells and the migration rate was as low as 33.4% in 20 μmol/L. Conclusion 1097C11 has good antitumor activity and is expected to be a lead compound targeting PLK1 relevant tumor.
出处
《中国医药生物技术》
2016年第1期13-20,共8页
Chinese Medicinal Biotechnology
基金
国家自然科学基金面上项目(81370087)
关键词
荧光偏振
抗肿瘤药
保罗样激酶1抑制剂
Fluorescence polarization
Antineoplastic agents
Polo-like kinase 1inhibitor
