RNA干扰技术用于survivin基因表达的研究  

Application of RNA interference on gene expression of survivin

作　　者：沈波[1] 鞠桂芝[1] 刘扬[1] 

机构地区：[1]吉林大学公共卫生学院放射生物学教研室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2006年第3期357-359,共3页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30270346)

摘　　要：目的:构建pSUPER-SVV表达载体,并在真核细胞中进行表达,验证survivin基因表达是否受到抑制。方法:设计特异性以survivin基因为靶序列的寡核苷酸序列,退火后将其连接于pSUPER.basic载体中,经测序鉴定插入序列的正确性。以脂质体转染方法将pSUPER-SVV干扰质粒转染人类宫颈癌(HeLa)细胞,应用流式细胞术和RT-PCR方法检测survivin基因的表达情况。结果:成功构建survivin的RNAi表达载体。该载体可以使HeLa细胞中survivin基因在mRNA转录水平明显降低,与转染空质粒组相比差异具有显著性(P<0.05)。利用流式细胞术检测蛋白表达水平,转染pSUPER-SVV组抑制率可达31.62%,与转染空质粒组相比差异具有显著性(P<0.001)。结论:成功构建干扰survivin基因表达的载体,转染HeLa细胞后survivin基因在mRNA转录水平、蛋白表达水平均受到明显抑制。Objective To construct pSUt）ER-SVV vector and obtain its expression in HeLa cells, in order to interfere the expression of survivin. Methods Oligonucleotide sequences specific for survivin were designed for RNA interference （RNAi）, after annealed they were ligated to pSUPER, basic vector. After sequencing, the vector of pSUPER-SVV was transfected into HeLa cells by LipofectAmine. The expression of survivin was detected by RT-PCR and FCM. Results The RNAi expression vector of survivin was constructed successfully, which depressed the expression of survivin efficiently. Survivin was significantly depressed on the level of mRNA in HeLa cells compared with the control groups （P〈0.05）, and the expression of survivin was down-regulated to 31.62%, there was significant difference compared with control group （P〈0.001）. Conclusion The RNAi vector targeted to survivin gene is successfully constructed, survivin is depressed on the levels of mRNA and protein in HeLa cells transfected by the vector.

关 键 词：SURVIVIN RNA干扰 基因表达 

分 类 号：Q78[生物学—分子生物学] R737.33[医药卫生—肿瘤]