RNAi技术对人肺腺癌细胞A549内源Survivin基因表达的影响  

Effects of RNA interference technology on the expression of endogenous Survivin in human lung adenocarcinoma cells A549

作　　者：董俊红[1] 王振明[2] 孙洪亮[1] 王平[1] 

机构地区：[1]潍坊医学院,山东潍坊261042 [2]潍坊市人民医院

出　　处：《山东医药》2009年第31期25-27,共3页Shandong Medical Journal

基　　金：山东省自然科学基金青年基金项目(Q2006D08)

摘　　要：目的为Survivin介导的肿瘤基因沉寂疗法的临床应用提供依据。方法应用RNA干扰技术(RNAi)将靶向Survivin的基因片段插入载体后构建重组质粒,将其导入A549细胞,MIT法检测转染前后A549细胞增殖情况,RT-PCR及Western blot法分析转染前后Survivin mRNA和蛋白表达情况。结果成功构建重组质粒pGene-sil1.1-Survivin。转染重组质粒后Survivin mRNA和蛋白的表达明显降低,抑制率分别为65%和71%,细胞增殖受抑。结论RNAi技术构建的重组质粒可明显抑制A549细胞内源Survivin的表达和mRNA转录,其在肿瘤基因沉寂疗法中可能起重要作用。Objective To provide experimental basis for the tumor gene silencing induced by Survivin. Methods The DNA sequence targeting human Survivin was inserted into the plasmid by RNA interference technology（ RANt）, the reeomhinant plasmid were constructed and transfected into A549 cells. Before and after transfection, the cell proliferation was observed by MTT, the levels of Survivin mRNA and Survivin protein were detected by RT-PCR and Western Blot. Results The recombinant plasmid pGenesill. 1-Survivin were successfully constructed. The A549 cells transfeeted with recombinant plasmid showed a lower mRNA and protein expression of Survivin, compared with th control group, the inhibition rate were 65% and 71% respectively. The A549 cell proliferation was inhibited. Conelusion The recombinant plasmid constructed by RNAi can inhibit the protein expression and RNA transcription of endogenous Survivin.

关 键 词：RNA干扰 SURVIVIN基因 人肺腺癌细胞 

分 类 号：R392.2[医药卫生—免疫学]