RNA干扰抑制神经母细胞瘤细胞MYCN基因的初步研究

Study on specific suppression of MYCN gene in neuroblastoma cells by small interfering

目的对RNA干扰抑制神经母细胞瘤细胞(LAN-5)MYCN基因表达进行初步研究。方法体外化学合成针对MYCN基因的小干扰RNA(si RNA),经脂质体Lipofectamine2000包裹后转染神经母细胞瘤细胞,以无关si RNA和未转染的细胞为对照,采用SYBR绿色荧光染料Ⅰ的实时荧光定量RT-PCR检测si RNA的抑制效果。结果脂质体转染si RNA效率可达84.83%,化学合成的si RNA可抑制神经母细胞瘤细胞MYCN基因mRNA表达,转染72h后抑制率达58.3%。结论化学合成的si RNA可以有效抑制神经母细胞瘤MYCN基因的表达,为神经母细胞瘤基因治疗开辟了新的途径。

Objective To investigate the specific suppression of MYCN gene in neuroblastoma Cells by small interfering RNA. Methods The small interfering RNA （siRNA） targeted to the MYCN mRNA was synthesized in vitro and was transfected by Lipofectamine 2002 into human neuroblastoma cell line LAN-5, which has high MYCN expression levels. The level of MYCN mRNA expression was quantified by quantitative reverse transcription polymerase chain reaction （RT-PCR） with SYBR GREEN I. Results The Lipofectamine 2002 transfection efficiency was 84.83%, the level of MYCN mRNA expression was significantly inhibited by siRNA. 72 hours after transfection, the suppression rate of MYCN mRNA expression was 58.3%. Conclusions The chemosynthesized MYCN siRNA can significantly down-regulate MYCN mRNA expression, which maybe a new way for neuroblastoma gene therapy.