RNA干扰沉默人血管平滑肌细胞胱抑素C基因表达及对组织蛋白酶S表达的影响

siRNA inhibits expression of Cystatin C in human vascular smooth muscle cells and effects expression of Cathepsin S

目的研究小干扰RNA(siRNA)对胱抑素C(CysC)基因表达的抑制作用及对组织蛋白酶S(CatS)表达的影响。方法利用Ambion公司设计合成的以CysC为靶标的siRNA,通过脂质体转染人血管平滑肌细胞(VSMC),以未转染siRNA细胞和转染无关siRNA细胞为对照,采用RT-PCR和Western blot分别检测CysC在mRNA水平和蛋白水平的改变。选取抑制效率最高siRNA转染VSMC,以空白对照组、无关干扰组和CysC高表达组为对照,采用RT-PCR法和Western blot分别检测CatS在mRNA水平和蛋白水平的改变。结果转染24 h后,siRNA组CysC mRNA和蛋白表达均被有效抑制,与空白对照组相比,差异有统计学意义(P<0.01),其中,siRNA2的抑制效率最高。CatS的mRNA及蛋白表达水平在siRNA组均有明显升高,与其他三组比较,差异有统计学意义(P<0.01)。结论体外转录合成的siRNA可有效抑制人VSMC中CysC的表达,并对CatS基因表达有一定影响。

Objective To evaluate the effect of small interference RNA(siRNA) on Cystatin C in human vascular smooth muscle cells and its effect on expression of Cathepsin S.Methods siRNA targeting Cystatin C was constructed by an online design of Ambion company and transfected into human vascular smooth muscle cells.The non-transfected cells and cells treated with the control siRNA served as controls.mRNA and protein expressions of Cystatin C were detected by RT-PCR and Western blot respectively.Compared with the non-...