靶向PRMT5先导化合物的筛选及其抗肿瘤作用探讨  

Screening of PRMT5-targeted compounds and study on their anti-tumor activities

作　　者：施晓柯[1] 李婕[2] 王任小[2] 潘景轩[1] 

机构地区：[1]中山大学中山医学院病理学与病理生理学教研室,广东广州510080 [2]中国科学院上海有机化学研究所生命有机化学国家重点实验室,上海200032

出　　处：《解剖学研究》2013年第3期200-202,224,共4页Anatomy Research

摘　　要：目的筛选可能作用于PRMT5蛋白的化合物并检测其抗肿瘤作用。方法以PRMT5蛋白为靶标,通过虚拟筛选技术挑选并购买得到101种化合物样品;以MTS法筛选所有化合物对人慢性粒细胞白血病K562细胞生长的抑制作用并计算IC50值;以化合物处理K562细胞48 h,收集细胞提取全蛋白,使用蛋白质印迹技术检测PRMT5下游甲基化蛋白H4RSDM的变化。结果以MTS法测试101种化合物,发现对K562细胞生长具有明显抑制作用的化合物共20个(其中8个化合物IC50<30μmol);蛋白质印迹技术进一步验证甲基化的H4RSDM蛋白表达水平并无明显变化,说明这些以PRMT5蛋白为靶标虚拟筛选获得的化合物并非通过抑制PRMT5甲基化来抑制K562细胞增殖的。结论所有测试的化合物中发现有8个对K562细胞的生长具有良好的抑制作用,但是其在分子水平上的作用机理还有待进一步研究。Objective To discover novel anti-tumor compounds by potentially targeting the PRMT5 protein.Methods A total of 101 compounds were purchased according to the outcomes of PRMT5-targeted virtual screening.These compounds were tested in MST assays to characterize their inhibitory effect on the growth of K562 cells.The active compounds were used to treat K562 cells at 60 μmol for 48 h.The cells were then lysed for Western blot experiments to detect the change in the level of methylated H4RSDM,one of the downstream signals of PRMT5 activation.Results A total of 101 compounds were selected by virtual screening targeting PRMT5 and were tested in MST assays.Twenty of them showed obvious inhibitory effect on K562 cell growth,among which eight compounds had IC50 30 μM.Western Blot experiments examining the level of methylated H4RSDM,however,demonstrated that the compounds inhibiting the growth of K562 cells did not affect PRMT5 activities.Conclusion We have discovered eight compounds that exhibit promising inhibitory effect on the growth of K562 cells.The molecular mechanism of these compounds however needs further investigations.

关 键 词：PRMT5蛋白 K562细胞系 H4RSDM甲基化蛋白 

分 类 号：R730.5[医药卫生—肿瘤]