CAJ | 학술논문

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Lipofectamine~(TM)2000 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Lipofectamine~(TM)2000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.
목적 연구siRNA 대대서신세포(NRK)Ppif 기인적억제작용.방법 근거대서Ppif 기인서렬설계병화학합성3대siRNA 급1대음성대조siRNA,병재5′단표기FAM 최기형광소,이Lipofectamine~(TM)2000 전염입대서신세포,용RT-PCR 화Western blot 분별검측Ppif 기인화단백표체량적개변,험증소설계적siRNA 적억제효응.결과이Lipofectamine~(TM)2000전염소설계적siRNA 진입NRK 세포,전염효솔＞90%.기시우405、556위점적siRNA 재기인수평대Ppif 무명현억제효응(P＞0.05),기시우198위점적siRNA 재기인화단백수평균유명현적억제효응,기인수평하강75.25%(P＜0.05);단백수평하강72.13 %(P＜0.05).결론 기시우198위점적siRNA 대대서신세포적Ppif 기인유명현적억제효응.
Objective To screen the siRNAs that 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 labeled 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.