目的 研究microRNA-183对神经母细胞瘤细胞的调控作用,为神经母细胞瘤的治疗提供新策略.方法 购买microRNA-183和对照microRNA,转染至人神经母细胞瘤细胞株SK-N-SH细胞,再使用MTT实验,Caspase-3活性测定试剂盒和流式检测细胞生长和凋亡的影响.合成Bcl-2 siRNA,检测Bcl-2对人神经母细胞瘤细胞株SK-N-SH细胞凋亡的影响.转染Bcl-2质粒后,再转染microRNA-183至人神经母细胞瘤细胞株SK-N-SH细胞,分析Bcl-2的表达水平和人神经母细胞瘤细胞株SK-N-SH细胞的凋亡.结果 转染microRNA-183降低SK-N-SH细胞的生长(P=0.0059),发生磷脂酰丝氨酸膜表面表达(P=0.008)和Caspase-3的激活(P=0.014),Bcl-2的表达降低(P=0.015).干扰SK-N-SH细胞中Bcl-2增强了microRNA-183诱导的细胞凋亡(P=0.0058),而过表达Bcl-2抑制了microRNA-183诱导的细胞凋亡(P=0.0073).结论 转染microR-NA-183抑制SK-N-SH细胞的生长和增殖.microRNA-183通过下调Bcl-2而诱导SK-N-SH细胞的凋亡,提示Bcl-2可能是神经母细胞瘤潜在的候选治疗靶点.%Objective To study the regulation of microRNA-183 on neuroblastoma cells and to provide a new strategy for the treatment of neuroblastoma.Methods microRNA and microRNA-183 were transfected into SK-N-SH cells,and then MTT experiment and Caspase-3 activity assay and flow cytometry were employed to investigate the role of microRNA-183 in growth and apoptosis of human neuroblastoma cell line SK-N-SH.Bcl-2 siRNA was transfected into SK-N-SH cells,and effects of Bcl-2 on hu-man neuroblastoma cell line SK-N-SH cells apoptosis was tested.After transfection of Bcl-2 plasmid,microRNA-183 was trans-fected into human neuroblastoma cell line SK-N-SH cells,and the expression level of Bcl-2 and apoptosis of human neuroblastoma cell line SK-N-SH were analyzed.Results The transfection of microRNA-183 reduced cell growth of the SK-N-SH cells (P=0.0059),and the expression of phosphatidylserine on the membrane (P=0.008)and activation of Caspase-3 were observed (P=0.014),and the expression of Bcl-2 was decreased (P=0.015).Bcl-2 interference of human neuroblastoma cell lines enhances ap-optosis of the human neuroblastoma cell line SK-N-SH induced by microRNA-183 (P=0.0058),and overexpression of Bcl-2 in-hibits apoptosis induced by microRNA-183 (P=0.0073).Conclusion The transfection of microRNA-183 inhibits the growth and proliferation of human neuroblastoma cell line SK-N-SH.MicroRNA-183 induces apoptosis of SK-N-SH by down regulating Bcl-2, which suggests that Bcl-2 may be a potential candidate therapeutic target for neuroblastoma.
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