saRNA上调人前列腺癌PC-3细胞中Numb基因表达的研究

Study on saRNA′s upregulation of Numb gene expression in human prostate carcinoma PC-3 cells

目的基于小激活RNA(saRNA)的肿瘤治疗策略,筛选具有激活Numb基因功能并相对高效的saRNA分子。方法设计与合成针对Numb基因的3对候选小分子双链RNA(dsRNA)分子,对照组dsRNA(dsControl)设计成与人类基因组序列非同源。将dsRNA分子转染人前列腺癌细胞(PC-3细胞)。采用RT-qPCR法检测转染后PC-3细胞中靶基因Numb的mRNA表达水平。Western blot验证转染后PC-3细胞靶基因Numb的蛋白表达水平。结果 dsNumb-870、dsNumb-948均未能上调PC-3细胞中靶基因Numb mRNA和蛋白水平;而dsNumb-298能上调细胞内Numb mRNA和蛋白水平。结论 dsNumb-298具有特异性激活PC-3细胞中Numb基因表达的功能。

Objective To screen the relatively high efficiency small activating RNA（saRNA）molecule possessing activating the target gene Numb based on the tumor treatment strategy of saRNA.Methods Three pairs of candidate small double-stranded RNA（dsRNA）molecules were designed and synthesized for target gene Numb,and the dsRNA in the control group（dsControl）was designed to be non-homologous with the human genome sequence.The dsRNA molecules were transfected into prostate carcinoma PC-3 cells.The real-time quantitative PCR（RT-qPCR）was employed to detect the mRNA expression levels of the target gene Numb in PC-3 cells after transfection.Then Western blot was used to verify the protein expression levels of the target gene Numb in PC-3 cells after transfecting the saRNA into the cells.Results dsNumb-870 and dsNumb-948 failed to up-regulate both the mRNA and protein expression levels of the target gene Numb in human prostate carcinoma PC-3 cells,while dsNumb-298 was succeed to elevate them significantly.Conclusion The dsNumb-298 molecule has the function to specially activate the gene Numb expression in human prostate carcinoma PC-3 cells.