小干扰RNA逆转三氧化二砷耐药的白血病K562／AS2细胞TopoⅡ基因表达的实验研究  

Inhibition effect of small hairpin RNA on expression of Topo lI gene in K562/AS2 cell line resistant to arsenic trioxide

作　　者：张旭艳[1] 卓家才[1] 杜新[1] 李明[1] 陶小梅[1] 史敦云[1] 楼瑾[1] 张琼丽[1] 

机构地区：[1]深圳市第二人民医院血液科深圳市血液病研究所,518035

出　　处：《白血病．淋巴瘤》2011年第6期357-361,共5页Journal of Leukemia & Lymphoma

基　　金：深圳市科技项目（200902052）

摘　　要：目的研究小干扰RNA片段（shRNA）对三氧化二砷（ATO）耐药的白血病细胞株K562／AS2细胞的TopoⅡα、TopoⅡβ基因表达及其功能的影响。方法设计并合成针对TopoⅡα和TopoⅡβ基因序列的shRNA各3对，在脂质体的介导下转染K562／AS2细胞；用荧光实时定量聚合酶链反应（PCR）分析TopoⅡα、TopopⅡβ mRNA的表达水平；流式细胞术检测TopoⅡα、TopoⅡβ蛋白表达。结果针对TopoⅡα—shRNA、TopoⅡβ的shRNA作用于K562／AS2细胞24h后，TopoⅡα mRNA水平和蛋白水平最大下调为（78．22±0．01）％、（31．17±1．27）％（P〈0．05），TopoⅡβ mRNA水平和蛋白水平最大下调为（57．36±0．01）％、（23．98±1．22）％（P〈0．05）。结论转染24h后针对TopoⅡ的shRNA可抑制对ATO耐药的白血病细胞株K562／AS2细胞TopoⅡ基因的表达。Objective To investigate the reversal effect of Topo Ⅱα-shRNA and Topo Ⅱβ-shRNA on Topo Ⅱ gene in K562/AS2 cells. Methods Three pieces of Topo Ⅱα-shRNA and three pieces of Topo Ⅱβ-shRNA were designed, synthesized and transfected into K562/AS2 cells by liposome. Expression level of Topo Ⅱα and Topo Ⅱβ mRNA were determined by real time fluorescence quantitative PCR. The expressions of Topo Ⅱα and TopoⅡβprotein were assayed with flow cytometer. Results After treated with Topo Ⅱα-shRNA or Topo Ⅱβ-shRNA for 24 hours, the expression level of Topo mRNA Ⅱαand Topo Ⅱβ mRNA protein in K562/AS2 cells decreased at most （78.22±0.01） %, （31.17±1.27） % （P 〈0.05）, and （57.36±0.01）%, （23.98±1.22） % （P 〈0.05） respectively. Conclusion The expression of Topo Ⅱ gene can be down-regulated after infected with Topo Ⅱ-shRNA in K562/AS2 cell line.

关 键 词：RNA 小分子干扰 DNA拓扑异构酶类 Ⅱ型 砷酸盐类 K562/AS2细胞 

分 类 号：R733.7[医药卫生—肿瘤]