摘要
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G_0/G_1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.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2011年第4期705-710,共6页
Chinese Journal of Pathophysiology
