T_7体外转录siRNA靶向干扰Pim-2效应观察  

Interfering Efficacy of siRNAs Synthesized in vitro with T_7 RNA Polymerase Targeted to hPim-2

作　　者：薛兴欢[1] 张淑群[1] 姜建涛[1] 王西京[1] 薛锋杰[1] 刘晓旭[1] 

机构地区：[1]西安交通大学医学院第二附属医院,陕西西安710004

出　　处：《中国肿瘤》2008年第4期305-308,共4页China Cancer

基　　金：陕西省科技攻关项目(2004K13-G9;2006K09-G9);国家自然科学基金(30500600)

摘　　要：[目的]观察T7RNA聚合酶介导体外转录合成siRNA靶向干扰Pim-2基因表达的效应。[方法]利用T7RNA聚合酶介导的体外转录合成靶向Pim-2的siRNAⅠ、Ⅱ、Ⅲ,将其转染入人结肠癌细胞株SW480。利用RT-PCR和Westernblot观察Pim-2基因在RNA和蛋白质水平的表达变化。[结果]与对照细胞相比,转染siRNAⅠ、Ⅱ、Ⅲ48h后,Pim-2基因mRNA的抑制率分别为65.4%(P<0.05),46.2%(P<0.05)和56.1%(P<0.05);转染72h后,Pim-2基因蛋白表达抑制率分别为61.6%(P<0.05),45.8%(P<0.05)和55.6%(P<0.05)。[结论]T7体外转录合成siRNA,能有效而特异地抑制人原癌基因Pim-2的表达。[Purpose] To investigate the efficacy performed by siRNAs synthesized in vitro with T7 RNA polymerase to modulate Pim-2 expression. [Methods] Colon cancer cells SW-480 were transfected with siRNAs Ⅰ , Ⅱ, Ⅲ targeted to hPim-2 and synthesized in vitro with T7 RNA polymerase. hPim-2 mRNA and protein expression were measured by RT-PCR and Western blot. [Results] After transfecing 48h with hPim-2 siRNA Ⅰ , Ⅱ, Ⅲ the inhibition rate of hPim-2 mRNA expression was 65.4% （P〈0.05）, 46.2%（P〈0.05） and 56.1% （P〈0.05） in colon cancer cells, respectively, compared to control cells. The inhibition rate of hPim-2 protein at 72h was 61.6% （P〈0.05）, 45.8% （P〈0.05） and 55.6% （P〈0.05）, respectively. [Conclusion] The siRNAs synthesized in vitro with T7RNA polymerase can be useful for silencing oncogene hPim-2 expression specifically and efficiently.

关 键 词：SIRNA PIM-2 干扰效应 

分 类 号：R737.5[医药卫生—肿瘤]