摘要
目的研究载体介导的siRNA(small interfere RNA)技术对骨肉瘤细胞株U2OS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U2OS细胞中MDM2-mRNA表达改变。结果成功构建发卡样MDM2siRNA真核表达载体(PGCsilencer TM-MDM2 siRNA);PGCsilencer TM-MDM2 siRNA转染到U2OS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PGCsilencerTM-MDM2 siRNA重组体,并能有效抑制U2OS细胞中MDM2-mRNA表达。
Objective investigate the effects of siRNA on MDM2-mRNA expression in U2OS cells. Methods The PGCsi- lencerTM-M：DM2 siRNA expression vector was constructed by gene recombination ,then transfected into the cultured U2OS cells. The expression of M：DM2 mRNA in U2OS were determined by semi-quantitative RT-PCR after the transfection. Results PC, Csilencer 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.
出处
《中国实验诊断学》
2007年第7期878-881,共4页
Chinese Journal of Laboratory Diagnosis
基金
吉林省科技厅发展计划项目资助课题(200502166)
