目的:探讨siRNA下调astrocyte elevated gene-1(AEG-1)表达对神经母细胞瘤细胞增殖、凋亡和细胞周期的影响.方法:用设计合成的AEG-1 siRNA转染神经母细胞瘤细胞株M17和SK-N-SH,采用荧光定量RT - PCR技术观察AEG-1 siRNA对AEG-1基因表达的影响;用MTT法和克隆形成实验观察AEG-1 siRNA对神经母细胞瘤细胞的增殖抑制作用;用流式细胞术检测下调AEG-1表达对神经母细胞瘤细胞凋亡及细胞周期的影响.结果:用AEG-1 siRNA转染人神经母细胞瘤细胞,RT - PCR结果证实AEG-1 siRNA能显著基因下调AEG-1表达,与对照组相比有显著差异(P<0.01);MTT法和克隆形成实验结果证明下调AEG-1表达可显著抑制神经母细胞瘤细胞的增殖;流式细胞术检测结果表明下调AEG-1表达可促进神经母细胞瘤细胞凋亡,并将细胞周期阻滞在G0/G1期.结论:AEG-1 siRNA可下调基因在神经母细胞瘤细胞中表达,并抑制神经母细胞瘤细胞增殖,促进细胞凋亡.%AIM: To investigate the effect of siRNA - induced astrocyte elevated gene - 1 ( AEG - 1) down -regulation on the proliferation, apoptosis and cell cycle of neuroblastoma cells.METHODS: An siRNA targeting to AEG - 1 mRNA ( AEG - 1 siRNA ) was constructed and transfected into neuroblastoma cells with Lipofectamine 2000.A non -specific siRNA ( control siRNA ) and non - treatment were used as negative control and blank control, respectively.The cell proliferation was detected by MTT method and colony formation assay.The apoptosis and cell cycle were analyzed by flow cytometry.RESULTS: Compared with control groups, the expression of AEG - 1 mRNA was evidently declined in the cells transfected with AEG - 1 siRNAs ( P ＜0.05 ).AEG - 1 siRNA significantly decreased the cell proliferation.After treated with AEG - 1 siRNA for 48 h, the percentage of apoptotic cells was significant increased and the cell cycle was arrested in G0/G1 phase compared with the control cells ( P ＜0.05 ).CONCLUSION: The mRNA expression of AEG - 1 is down - regulated by AEG - 1 siRNA in neuroblastoma cells.Knockdown of AEG - 1 expression in human neuroblastoma cells significantly inhibits cell proliferation and apoptosis, and induces cell arrest in G0/G1 phase of the cell cycle.
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