【摘要】 目的筛选高效dsRNA/mdr1,以备研究RNA干扰逆转肝细胞癌多药耐药之用。方法首先根据siRNA设计原则,以多药耐药基因（mdr1）为靶基因,设计并选择4⁵条siRNA/mdr1,经BLAST后体外转录法合成dsRNA/mdr1,用Oligofectamine试剂分别转染HepG2/mdr1,然后从mRNA、蛋白（P-gp）表达水平和细胞功能变化评价HepG2/mdr1耐药性被逆转的程度,比较各个dsRNA/mdr1的逆转效率,筛选出有效的siRNA/mdr1。结果成功合成5条dsRNA/mdr1（其中1条为阴性对照）,dsRNA/mdr1-4mRNA表达（18.73±1.33）%、蛋白表达变化（79.1±1.6）%^16.8±0.4%与其他各组细胞比较,有显著性差异;细胞内柔红霉素（DNR）累积量也较其他组明显增加（平均荧光强度79.58,阳性率84.25%,P<0.05）。结论体外转录法结合脂质体转染适用于筛选高效siRNA,肯定了siRNA干扰序列能够有效阻抑mdr1基因编码蛋白p170的功能。更多还原

【Abstract】 Objective To screen effective dsRNA/mdr1 for studying the reversal of multidrug resistance of hepatocellular carcinoma (HCC) by RNA interference (RNAi). Methods dsRNA/mdr1 targeting multidrug resistance gene (mdr1) was designed and synthesized by in vitro transcription. HepG2/mdr1 in 6 groups was transfected with the complex of different dsRNA/mdr1 (dsRNA/mdr1-1, dsRNA/mdr1-2, dsRNA/mdr1-3, dsRNA/mdr1-4, dsRNA/mdr1-5) using Oligofectamine. Then the cells were collected to measure the expression of mRNA/mdr1, P-glycoprotein and the accumulation of DNR. Results Five dsRNA/mdr1 were successfully synthesized including one negative. The expression of mRNA/mdr1 and P-glycoprotein (from (79.1±1.6)% before transfection down to (16.8±0.4)%] in dsRNA/mdr1-4 was significantly lower than that of other groups (P<0.05). Accumulation of DNR in dsRNA/mdr1-4 was higher than that in other groups (P<0.05). Conclusion In vitro transcription combined with liposome is fit to screen the effective dsRNA/mdr1. siRNA/mdr1 could suppress the expression of P-glycoprotein coded by mdr1 gene.更多还原