磷硫代siRNA的YAP基因沉默活性研究

Study on YAP gene silencing activity of phosphorothioate siRNAs

为了探究RNA聚合酶酶促合成非对映体纯PS-siRNA(pPS-siRNA)的基因沉默活性,本研究针对潜在的癌症治疗靶点Yes相关蛋白(YAP),通过qPCR、Western blot等方法研究了其对YAP基因的沉默效果和对HeLa细胞的细胞毒性.结果表明,与未修饰的siRNA(PO-siRNA)相比,pPS-siRNA可以更好地下调YAP基因表达(效率提高约30%),而没有明显的细胞毒性.此外,MTT细胞增殖实验与流式细胞术的结果表明,经pPS-siRNA敲降YAP后,HeLa细胞的增殖受到了抑制.说明pPS-siRNA在基因治疗方面具有优越性以及YAP作为肿瘤治疗靶点的潜力.

In order to explore the gene silencing activity of diastereomeric pure PS-siRNA(pPS-siRNA)synthesized enzymatically by RNA polymerase,its silencing activity on Yes-associated protein(YAP),a potential cancer treatment target,and cytotoxicity to HeLa cells were studied by qPCR and Western blot in this study.The data showed that compared with unmodified siRNAs(PO-siRNAs),pPS-siRNAs can better reduce YAP gene expression without significant cytotoxicity(approximately 30% more efficient).In addition,MTT cell proliferation assay and flow cytometry showed that the proliferation of HeLa cells was inhibited after YAP down-regulated by the pPS-siRNAs.This research proved the superiority of pPS-siRNAs in gene therapy and the potential of YAP as a tumor therapeutic target.