RNA干扰技术对PC12细胞CaMKⅡβ基因表达的影响

Inhibition of CaMKⅡβ Expression by RNA Interference in PC12

观察RNA干扰表达载体对PC12细胞中CaMKⅡβ基因的抑制作用。构建针对CaMKⅡβ基因的RNAi质粒表达载体,脂质体法转染大鼠肾上腺嗜铬细胞瘤PC12细胞株,应用逆转录-聚合酶链反应(RT-PCR)、Western blot检测其对CaMKⅡβ mRNA及蛋白质水平的影响。构建的质粒表达载体在PC12细胞中抑制了CaMKⅡβ mRNA及蛋白质的表达。与阴性对照组相比,表达质粒产生的siRNA对CaMKⅡβ mRNA的抑制率48h,72h分别为46.40%,64.69%,蛋白抑制率72h约为74.77%。RNAi表达载体可以有效地抑制PC12细胞CaMKⅡβ基因的表达。

To investigate the inhibitory effects of RNAi cell lines, siRNA expression vectors were constructed to be expression vector on CaMKIIβ expression of PC12 targeted directly at CaMKIIβ gene. The recombinants were transfected into PC12 cell line with liposome. The expression of CaMKIIβ was detected by RT-PCR and Western blot. Expression vectors could reduce the expressions of CaMKIIβ mRNA and protein in PC12, Compared with blank vector group, the former, the ratio of inhibition of the expression of CaMKIIβ mRNA was 46.40% and 64.69%, in 48th hour and 72th hour, the ratio of inhibitory of the expression of CaMKIIβ protein was 74.77% in 72th hour. RNAi expression vectors can effectively inhibit the expression of CaMKIIβ in PC12.