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目的观察Musashi(Msi)2对急性髓系白血病(acute myelogenous leukemia,AML)THP-1细胞体外增殖能力的影响,初步探讨Msi2在AML中可能的调控作用。方法利用RNA干扰技术沉默Msi2表达,实时荧光定量PCR法检测Msi2、cyclin D1、p21、cdk2基因mRNA转录水平;Western blot法检测Msi2蛋白表达水平;生长曲线观察THP-1细胞体外增殖能力;流式细胞仪检测细胞周期进程。结果与scramble组及空白对照组相比,Msi2-siRNA组细胞cyclin D1、cdk2基因mRNA转录水平明显降低(P<0.05),p21基因mRNA转录水平明显增高(P<0.05),Msi2 mRNA转录及蛋白表达水平均明显降低(P<0.05);Msi2-siRNA组细胞体外增殖能力降低(P<0.01);G1期细胞比例明显增高(P<0.05),S期细胞比例显著降低(P<0.05)。结论 Msi2可能通过调控细胞周期进程,促进白血病细胞恶性增殖,在AML的发生发展中发挥重要作用。
Objective To investigate the effect of Musashi (Msi) 2 on proliferation of acute myelogenous leukemia (AML) THP-1 cells in vitro and to explore the possible regulatory role of Msi2 in AML. Methods Msi2 expression was silenced by RNAi technique. Msi2, cyclin D1, p21 and cdk2 mRNA expressions were detected by real-time fluorescence quantitative PCR. The expression of Msi2 protein was detected by Western blotting. Growth curve was used to observe the proliferation of THP-1 cells in vitro. Cytometry detects cell cycle progression. Results Compared with the scramble group and blank control group, the mRNA transcription levels of cyclin D1 and cdk2 in Msi2-siRNA group were significantly decreased (P <0.05), the mRNA transcription level of p21 gene was significantly increased (P <0.05), Msi2 mRNA transcription and protein (P <0.01). The proportion of cells in G1 phase was significantly increased (P <0.05), while the proportion of cells in S phase was significantly decreased (P <0.05). Conclusion Msi2 may play an important role in the development of AML by regulating the cell cycle progression and promoting the malignant proliferation of leukemia cells.