VEGFR1基因特导性siRNA的筛选

目的]筛选血管内皮生长因子受体(VEGRF)基因特异性小干扰RNA(Small Interference RNA,siRNA),为肿瘤等疾病的基因治疗寻找一种新途径。方法]以高表达VEGFR1的人脐静脉血管内皮细胞(HUVEC)为模型,采用RNA干扰技术,化学合成了3条针对血管内皮生长因子受体1(VEGFR1)的特异性siRNA,用Lipofectamine2000TM转染HUVEC细胞株,通过Real time RT-PCR技术检测HUVEC细胞VEGFR1基因mRNA的表达。并对效果最好的VEGFR1 siRNA-2进行siRNA的浓度梯度效果检测。结果]结果表明,与对照组相比,所设计的3条siRNA均能不同程度地抑制VEGFR1 mRNA的表达,其中siRNA-2号最有效,浓度为50 nmol/L时抑制率达到95%左右。在浓度梯度试验中,VEGFR1 siRNA-2转染浓度为50 pmol/L时,对VEGFR1基因的沉默效果还能达到50%左右。结论]所设计的siRNA能有效抑制VEGFR1基因的表达,为RNAi用于靶向VEGFR1的基因治疗提供了非常有效的siRNA序列。

Screening of VEGFR1 specific siRNA for therapeutic application

Objective] The small interference RNAs(siRNAs) of vascular endothelial growth factor receptor 1(VEGFR1) was selected for a the new approach of tumor and other diseases-curing.Method] In this study,we have applied RNA interference(RNAi) technology by utilized chemically synthesized small interference RNAs(siRNAs) to block the gene expression of vascular endothelial growth factor receptor 1(VEGFR1) in human umbilical vein endothelial cells(HUVEC).The initial experiments were carried out with 50 nM various siRNAs to determine the expression of VEGFR1 mRNA by Real time RT-PCR.Result The results demonstrated that VEGFR1 siRNA-2 potently suppressed the expression of VEGFR1 mRNA in HUVEC.In attempt to identify the siRNA interfering efficiency,we have performed with a various concentrations of siRNA: 50 nmol/L,20 nmol/L,10 nmol/L,1 nmol/L,500 pmol/L,250 pmol/L,100 pmol/L,50 pmol/L.We observed that even in the concentration of 50 pM,siRNA-2 can effectively inhibit the expression of VEGFR1 mRNA. Conclusion] VEGFR1 siRNA-2 specifically suppresses the VEGFR1 expression,and may have the potential as an effective siRNA sequence in tumor gene therapy targeting VEGFR1.