摘要
目的研究siRNA对大鼠肾细胞(NRK)Ppif基因的抑制作用。方法根据大鼠Ppif基因序列设计并化学合成3对siRNA及1对阴性对照siRNA,并在5′端标记FAM羧基荧光素,以LipofectamineTM2000转染入大鼠肾细胞,用RT-PCR和Western blot分别检测Ppif基因和蛋白表达量的改变,验证所设计的siRNA的抑制效应。结果以Li-pofectamineTM2000转染所设计的siRNA进入NRK细胞,转染效率>90%。起始于405、556位点的siRNA在基因水平对Ppif无明显抑制效应(P>0.05),起始于198位点的siRNA在基因和蛋白水平均有明显的抑制效应,基因水平下降75.25%(P<0.05);蛋白水平下降72.13%(P<0.05)。结论起始于198位点的siRNA对大鼠肾细胞的Ppif基因有明显的抑制效应。
Objective To screen the siRNAs thai could inhibit the expression of peptidylprolyl isomerase F（Ppif） in the normal rat kidney（NRK）cells. Methods Three Ppif siRNAs and one negative control siRNA were designed, synthesized and la beled with FAM carboxyfluorescein at the 5＇ end for the measurement of transfection efficiency, siRNAs were transfected into NRK cells with Lipofectamine^TM 2000. RT-PCR and Western blot were performed to detect the expression of Ppif mRNA and protein, respectively. Results The transfection efficiency of the three siRNAs and the negative control siRNA was all more than 90%. The siRNAs at the 405 and 556 gene positions couldn＇t significantly inhibit the expression of Ppif（P〉0.05）. However, the siRNA at the 198 gene position significantly decreased the expression of Ppif mRNA and protein by 75.25%（P〈0.05）and （72.13%）（P〈0.05）respectively. Conclusion The siRNA at the 198 gene position could effectively inhibit the expression of Ppif in NRK cells.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2010年第1期98-101,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.30670820)
湖北省科技攻关计划资助项目(No.2006AA301B52-7)
