VEGFR1基因特异性siRNA的筛选

Screening of VEGFR1 specific siRNA for therapeutic application

[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。

[ Objective] The small interference RNAs （siRNAs） of vascular endothelial growth factor receptor 1 （VEGFRI） 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 （VEGFRI） in human umbilical vein endothelial cells （HUVEC）, The initial experiments were carried out with 50 nM various siRNAs to determine the expression of VEGFRI 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 VEGFRI mRNA, [ Conclusion] VEGFRI siRNA_ 2 specifically suppresses the VEGFR1 expression, mad nr：ty have the potential as an effective siRNA sequence in tumor aene therapy targeting VEGFR1.