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目的探讨丙戊酸钠(VPA)对多发性骨髓瘤(MM)细胞株RPMI8226细胞Notch受体及配体表达的影响。方法培养RPMI8226细胞后,分为4组,对照组(空白)和小中大3个浓度(VPA 2,4,8 mmol·L-1)实验组。用不同浓度VPA处理RPMI8226细胞24,48,72 h,用四甲基偶氮唑蓝比色法检测细胞增殖抑制作用。用反转录聚合酶链式反应、蛋白免疫印迹方法检测细胞Notch1、jagged1及jagged2 mRNA和Notch1、jagged1及jagged2蛋白在VPA作用后48 h的表达。结果 VPA对RPMI8226细胞的增殖抑制作用具有明显的浓度-时间依赖效应。不同浓度VPA作用RPMI8226细胞48 h,与对照组相比,Notch1、jagged1、jagged2 mRNA和Notch1、jagged1、jagged2蛋白表达均明显降低(P<0.05),且随着VPA浓度的不断增高,各基因和蛋白的表达量逐渐降低,3个浓度VPA组间两两比较,差异均有统计学意义(P<0.05)。结论 VPA能够下调RPMI8226细胞Notch1受体、jagged1及jagged2配体的表达水平,这可能是VPA治疗多发性骨髓瘤的作用机制之一。
Objective To investigate the effects of valproate (VPA) on the expression of Notch receptor and ligand in multiple myeloma (MM) cell line RPMI8226. Methods After RPMI8226 cells were cultured, they were divided into 4 groups: control group (blank) and small, medium and large groups (VPA 2,4,8 mmol·L-1). RPMI8226 cells were treated with different concentrations of VPA for 24, 48 and 72 h, and the cell proliferation inhibition was detected by MTT assay. The expression of Notch1, jagged1 and jagged2 mRNA and Notch1, jagged1 and jagged2 protein in 48 h after VPA treatment were detected by reverse transcriptase polymerase chain reaction and Western blot. Results VPA had a significant concentration-time-dependent effect on the proliferation of RPMI8226 cells. Compared with the control group, the expression of Notch1, jagged1, jagged2 mRNA and Notch1, jagged1, jagged2 protein in RPMI8226 cells were significantly decreased at different concentrations of VPA (P <0.05), and with the increasing of VPA concentration, The protein expression decreased gradually. There was a significant difference (P <0.05) between the three concentrations of VPA in every pairwise comparison. Conclusion VPA can down-regulate the expression of Notch1 receptor, jagged1 and jagged2 ligand in RPMI8226 cells, which may be one of the mechanisms of VPA in the treatment of multiple myeloma.