RNAi抑制前列腺癌雄激素受体表达的实验研究

目的:利用RNA干扰(RNAi)效应抑制雄激素受体(AR)的表达,观察RNAi在人前列腺癌PC3细胞中的干扰效应,寻找高效率的干扰片断。方法:设计5个针对AR的不同的siRNA(siRNA1、siRNA2、siRNA3、siRNA4和siRNA5)为实验组,构建表达载体并瞬时转染PC3细胞,提取总RNA和总蛋白,分别行荧光定量PCR和Western印迹检测AR mRNA和蛋白的表达,另设一无意义RNA表达载体为阴性对照组,比较实验组与对照组间的差异。结果:各实验组的AR mRNA的表达较对照组均不同程度下降,统计学分析差异有显著性(P(0.05),以siRNA1、siRNA4和siRNA5抑制效率最高,对比其余实验组差异有显著性(P(0.05),AR蛋白检测显示相同结果。结论:RNAi技术可有效地抑制前列腺癌细胞AR的表达,找到了具有较高抑制效率的siRNA并合成了其表达载体,为下一步的体内实验和药物研制奠定了基础。

siRNA Knocks Down the Expression of AR Gene in PC3 Cell Lines

OBJECTIVE: To observe the suppression of the expression of androgen receptor (AR) gene in PC3 cells after AR-specific siRNAs transfection, and to search for the siRNA (s) with the greatest suppressing efficiency. METHODS: Five AR-specific siRNAs were selected, RNAi expression vectors were constructed and transfected into PC3 cells, and the AR expression was detected by real time FQ-PCR and Western blot. A nonsense small RNA was set as negative control. RESULTS: Compared with the control group, the AR expression decreased in various degrees in the 5 experimental groups (P < 0.05), and siRNA1, siRNA4 and siRNA5 showed the greatest suppressing efficiency as compared with the other experimental groups, with statistically significant difference (P < 0.05). CONCLUSION: The AR-specific siRNAs could suppress the endogenous expression of target gene. Three siRNAs with great suppressing efficiency were identified and the expression vectors were constructed successfully. It can be applied in the future researches in vivo.