青藤碱类衍生物联合二甲双胍诱导骨髓瘤细胞凋亡的实验研究

Experimental study of sinomenine derivative combined with metformin on induction of apoptosis in multiple myeloma cells

目的探究青藤碱类衍生物YL064联合二甲双胍对多发性骨髓瘤(multiple myeloma,MM)细胞凋亡的生物学效应。方法以不同浓度的YL064和二甲双胍单独或联合处理MM细胞系的RPMI8226和H929细胞,采用细胞计数试剂盒-8检测细胞增殖抑制率,应用药物联合指数(combination index,CI)分析两药是否存在协同效应,同时应用流式细胞仪检测细胞凋亡率,进一步利用蛋白质印迹法检测细胞凋亡相关蛋白聚ADP核糖聚合酶(poly ADP-ribose polymerase,PARP)和caspase-3表达的变化。结果RPMI8226和H929细胞经二甲双胍和YL064单独或联合处理24 h后,联合用药组细胞增殖抑制率均显著高于单独用药组,联合用药组RPMI8226和H929细胞的CI均小于1。联合用药组RPMI8226和H929细胞凋亡率亦显著高于单独用药组,差异有统计学意义(P<0.05)。进一步研究显示,联合用药组RPMI8226和H929细胞内PARP和caspase-3均发生明显剪切活化。结论青藤碱类衍生物YL064联合二甲双胍能够协同诱导MM细胞增殖阻滞并进一步诱导其凋亡。

Objective To investigate possible effects of sinomenine derivative YL064 combined with metformin on multiple myeloma (MM) cells.Methods The MM cell lines RPMI8226 and H929 cells were treated with different concentrations of YL064 and metformin alone or two drugs combination for 24 h.The proliferation of RPMI 8226 and H929 cells were evaluated through cell counting kit-8 assay.The synergistic effects of two agents were calculated by combination index (CI).Meanwhile,flow cytometry was used to analyze changes of cell apoptosis rate.Furthermore,the expression of poly ADP-ribose polymerase (PARP) and caspase-3 were detected by western blot.Results After treating with YL064 and/or metformin,the growth inhibition rate of RPMI8226 and H929 cells in combination groups both significantly higher than those in YL064 or metformin alone group.The CI of YL064 combined with metformin was less than 1 in RPMI8226 and H929 cells.The apoptosis rate of RPMI8226 and H929 cells in combination groups both significantly higher than those in YL064 or metformin alone group,with statistical significant(P<0.05).Further study showed that YL064 synergized with metformin,to induce degradation of caspase-3 and PARP in RPMI8226 and H929 cells.Conclusions Sinomenine derivative YL064 could synergically induce proliferation inhibition and apoptosis in MM cells.