阿扎胞苷诱导多发性骨髓瘤细胞株的凋亡及其机制的研究

Inductive Effect of 5-Azacitidine on Apoptosis of Multiple Myeloma Cell lines and Its Mechanism

目的:探讨阿扎胞苷对多发性骨髓瘤细胞株U266和H929的抑制增殖、诱导凋亡的作用及其机制。方法:采用CCK-8分析阿扎胞苷对多发性骨髓瘤细胞株的增殖的抑制作用,应用吖啶橙染色和流式细胞术分析细胞DNA含量以判断阿扎胞苷对多发性骨髓瘤细胞株诱导凋亡作用,流式细胞术分析细胞周期以判断阿扎胞苷对多发性骨髓瘤细胞株细胞周期的影响,RT-PCR检BCL-2、BAX的表达,Western blot检测caspase-3和p-ERK1/2性以分析阿扎胞苷诱导骨髓瘤细胞凋亡的可能机制。结果:阿扎胞苷抑制多发性骨髓瘤细胞的增殖,导致多发性骨髓瘤细胞BCL-2/BAX比例下降,caspase-3和p-ERK1/2活性增高,使多发性骨髓瘤细胞周期停止在G_2/M期,诱导多发性骨髓瘤细胞凋亡且呈剂量依赖性。结论:阿扎胞苷对多发性骨髓瘤细胞具有抑制增殖和诱导凋亡作用,其作用机制可能与BCL-2/BAX比例下降,caspase-3活化以及影响细胞周期有关。

Objective:To investigate the response of multiple myeloma(MM) cells to 5- azacitidine and its possible mechanisms.Methods:Two multiple myeloma- derived cell lines U266 and H929 were used in this study.The cell proliferation and apoptosis were assessed by CCK-8,flow cytometry and acridine orange staining.The cell cycle was assessed by flow cytometry.The expression of BCL-2,BAX was assessed by RT-PCR.The expressions of caspase-3 and p-ERK1/2 were assessed by Western blot.Results:The BCL-2/BAX ratio decreased,the activity of caspase-3and p-ERK1/2 increased,the cell cycle was arrested in G_2 /M phase after treatment with 5-azacitidine.The 5-azacitidine inhibited proliferation in a dose-dependent manner.Conclusion:5-azacitidine exerts apoptosis- inducing and growinhibiting effects on MM cell lines,its mechanism may be related with the decrease of BCL-2/BAX ratio,caspase-3activation and the arrest of cell cycle.