艾拉莫德对人多发性骨髓瘤细胞株RPMI8226增殖和凋亡的影响

Effects of iguratimod on proliferation and apoptosis of human multiple myeloma cell line RPMI8226

目的探讨艾拉莫德对人多发性骨髓瘤(multiple myeloma,MM) RPMI8226细胞增殖和凋亡的影响。方法实验分组为:对照组(0μg/ml艾拉莫德),10μg/ml艾拉莫德处理组,20μg/ml艾拉莫德处理组,30μg/ml艾拉莫德处理组。艾拉莫德(10,20,30μg/ml)作用于人多发性骨髓瘤细胞株RPMI8226细胞24 h,采用MTT法检测RPMI8226细胞活力,Hoechst 33258染色检测凋亡细胞;荧光定量PCR检测Notch1 m RNA表达水平; Western blot检测Bax和Notch1蛋白的表达水平。结果10,20和30μg/ml的艾拉莫德处理24 h,可降低RPMI8226细胞活力,并具剂量依赖性(P <0. 05)。艾拉莫德(10,20,30μg/ml)处理RPMI8226细胞24 h,细胞核出现固缩、裂解现象。10,20和30μg/ml的艾拉莫德处理细胞24 h可显著上调RPMI8226细胞的Bax蛋白表达水平(P <0. 05),并显著下调Notch1 m RNA和蛋白水平(P <0. 05)。结论艾拉莫德可抑制人多发性骨髓瘤细胞株RPMI8226细胞增殖并促进细胞凋亡,其作用机制可能与其抑制Notch通路的活化有关。

Objective To explore the effects of iguratimod on cell proliferation and apoptosis of human multiple myeloma cell line RPMI8226 cells. Methods The experiment was divided into control group(0 μg/ml iguratimod), 10 μg/ml iguratimod group, 20 g/ml iguratimod group, and 30 μg/ml iguratimod group. After treated RPMI8226 cells with different concentrations of iguratimod for 24 h, the cell viability was measured using MTT assay, the morphology of apoptotic cell was observed under a fluorescence microscope after Hoechst 33258 staining, the expression level of Notch1 mRNA was detected by quantitative real-time PCR, and the expression levels of Bax and Notch1 protein were detected by Western blot. Results Compared with control group, iguratimod significantly reduced the viability of RPMI8226 cells in a dose dependent manner( P <0.05). The cell nuclei showed pyknosis and lysis in RPMI8226 cells treated with iguratimod(10, 20, and 30 μg/ml) for 24 h, and the expression level of Bax protein was upregulated( P <0.05). In addition, the levels of Notch1 mRNA as well as protein in RPMI8226 cells were markedly ameliorated after treatment with iguratimod(10, 20, and 30 μg/ml) for 24 h( P <0.05). Conclusion Iguratimod can inhibit cell proliferation and induce cell apoptosis of human myeloma cell line RPMI8226,which may be related to inhibition of Notch pathway activation.