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为了探究位于IGF1R基因3’UTR与前列腺癌(prostate cancer,PCa)相关的多态性位点rs2016347通过改变与miRNA结合影响PCa的风险,本研究用miRbase预测靶向miRNA,然后用热力学模型方法计算结合能情况,最后用双荧光素酶报告基因技术对HEK293T在体外检测改变rs2016347位点对靶向miRNA结合的影响。miRbase预测结果显示,hsa-miR-3175与IGF1R的结合区位于多态性位点rs2016347。热力学模型方法计算结果表明,相对位点G,hsa-miR3175与IGF1R的3’UTR端在rs2016347位点T上更能有效结合。双荧光素酶报告基因的检测结果显示,hsa-miR-3175能与带有rs2016347等位位点(G或T)IGF1R 3’UTR结合,hsa-miR-3175与等位位点T的结合更稳定,hsa-miR-3175与IGF1R基因的结合起到稳定IGF1R基因表达的作用。多态性位点rs2016347等位位点T在PCa的发展中风险性更大。
In order to explore the polymorphism of rs2016347 in the 3’UTR and prostate cancer (PCa) of IGF1R gene, we predicted the targeted miRNA by miRbase by changing the risk of PCa binding with miRNA, and then calculated it by thermodynamic model Finally, dual luciferase reporter gene technology was used to detect the effect of rs2016347 on target miRNA binding in HEK293T cells in vitro. The results of miRbase prediction showed that the binding region between hsa-miR-3175 and IGF1R was located at the polymorphic site rs2016347. The results of the thermodynamic model method show that the relative position of G, hsa-miR3175 and IGF1R 3’UTR end at rs2016347 site T more effective binding. The result of dual luciferase reporter assay showed that hsa-miR-3175 could bind to IGF1R 3’UTR with rs2016347 allele (G or T), and the binding of hsa-miR-3175 to allele T was more Stable, the binding of hsa-miR-3175 to the IGF1R gene plays a role in stabilizing IGF1R gene expression. Polymorphic site rs2016347 allele T is more risky in the development of PCa.