艾拉莫德通过活化AMPK对骨髓瘤细胞株U266增殖的影响

Effects of iguratimod on proliferation of myeloma cell line U266 by regulating AMPK activation

目的:探讨艾拉莫德对多发性骨髓瘤(multiple myeloma,MM)细胞株U266细胞增殖的影响。方法:艾拉莫德(10、20和30μg/ml)作用于骨髓瘤细胞株U266细胞后,采用四甲基偶氮唑蓝(MTT)法检测U266细胞活力;细胞周期染色试剂盒检测U266细胞周期分布情况; Western blot检测细胞周期相关蛋白CDK2、AMPK及p-AMPK的表达水平。结果:10、20和30μg/ml的艾拉莫德处理24 h后,U266细胞活力呈剂量依赖性的降低。艾拉莫德(10、20和30μg/ml)处理U266细胞24 h后,G_0/G_1期细胞数明显增加,S期细胞数明显降低。本研究还进一步发现10、20和30μg/ml的艾拉莫德处理24 h后可显著下调U266细胞的CDK2蛋白表达水平。此外,艾拉莫德人处理骨髓瘤细胞株U266细胞24 h后,U266细胞的p-AMPK/AMPK水平明显增加。结论:艾拉莫德可抑制骨髓瘤细胞株U266细胞增殖以及细胞周期,其作用机制可能涉及促进AMPK的活化。

Objective To explore the effects of iguratimod on cell proliferation of multiple myeloma（MM）cell line U266 cells. Methods： After treating U266 cells with different concentrations of iguratimod （10,20,30 μg/ml） for 24 h,cell viability was measured using MTT assay.The distribution of cell cycle was determined by flow cytometry （FCM） using Cell cycle staining Kit.The expression levels of CDK2,AMPK and p-AMPK protein were detected by Western blot. Results： Compared with the control group,iguratimod significantly reduced the viability of U266 cells in a dose dependent manner.Furthermore,administration of U266 cells with iguratimod （10,20,30 μg/ml） for 24 h,the numbers of U266 cells in G 0/G 1 period were significantly enhanced while the cell numbers in S period were significantly decreased.In the present study,we also found that the expression level of CDK2 protein was downregulated.Additionally,the level of p-AMPK/AMPK in U266 cells were obviously elevated after treatment with 20 or 30 μg/ml iguratimod for 24 h. Conclusion： Iguratimod can inhibit cell proliferation as well as cell cycle of myeloma cell line U266 and the underlying mechanism may be associated with promotion of AMPK activation.