siRNA对骨肉瘤U2OS细胞株MDM2-mRNA表达的抑制作用

目的 研究载体介导的siRNA（small interfere RNA）技术对骨肉瘤细胞株U20S中MDM2-mRNA表达的抑制作用.方法 依据设计siRNA的原则,以MDM2为靶基因设计并合成有小发夹结构的两条DNA序列,经退火成互补双链,再克隆到载体PGCsilencerTM中构建重组体（PGCsilencerTM-MDM2 siRNA）.实验分转染重组质粒组,转染阴性对照质粒组和只加脂质体的空白组;RT-PCR法观察U20S细胞中MDM2-mRNA表达改变.结果 成功构建发卡样MDM2 siRNA真核表达载体（PGCsilencer TM-MDM2 siRNA）;PGCsilencer TM-MDM2 siRNA转染到U20S细胞后,MDM2-mRNA表达较空白组显著下降（P＜0.05）,转染阴性对照质粒组与空白组无显著差异（P＞0.05）.结论 成功构建PGCsilencer TM-MDM2 siRNA重组体,并能有效抑制U20S细胞中MDM2-mRNA表达.

Inhibition effects of siRNA on MDM2-mRNA expression in U2OS cells

Objective investigate the effects of siRNA on MDM2mRNA expression in U2OS cells.Methods The PGCsilencerTM-MDM2 siRNA expression vector was constructed by gene recombination,then transfected into the cultured U2OS cells.The expression of MDM2 mRNA in U2OS were determined by semi-quantitative RT-PCR after the transfection.Results PGCsilencer TM-MDM2 siRNA expression vector was successfully constructed.Transfection of PGCsilencerTM-MDM2 siRNA expression vector into U2OS cells down-regulated the mRNA expression level of MDM2 gene and as compared with the untreated group,P<0.05.There is no remarkable difference between control group and untreated group,P>0.05.Conclusion The PGCsilencer TM-MDM2 siRNA expression vector can inhibit the expression of MDM2-mRNA in U2OS cells.