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靶向PTTG和survivin基因共干扰质粒对U251细胞的沉默效率 被引量:2

Silence effects of double interfering vector targeting PTTG and survivin genes on U251 cells
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摘要 目的:探讨共干扰质粒pGenesil-2.1-PTTG-survivin siRNA对U251细胞人垂体瘤转化基因(PTTG)和生存素(survivin)基因表达的影响。方法:U251细胞分为5组,分别为正常细胞对照组、Lipo2000+pGenesil-2.1HK组(HK阴性对照组)、Lipo2000+pGenesil-2.1-PTTG siRNA、Lipo2000+pGenesil-2.1-survivin siRNA及Lipo2000+pGenesil-2.1-PTTG-survivin siRNA。应用RT-PCR法、流式细胞术和Westernblotting方法检测转染36h后的各组U251细胞PTTG与survivin基因mRNA和蛋白表达。结果:3个干扰组的U251细胞中PTTG和survivin基因mRNA和蛋白表达水平均较HK阴性对照组显著降低(P<0.01),以共干扰质粒pGenesil-2.1-PTTG-survivin siRNA组降低最明显。结论:共干扰pGenesil1-PTTG-survivin siRNA质粒对U251细胞中PTTG与survivin基因的沉默效应强于单独沉默其中某一基因的干扰质粒。 Objective To explore the effects of double interfering vector pGenesil-2.1-PTTG-survivin siRNA on PTTG and survivin gene expressions in human U251 glioma cell line.Methods U251 cells were divided in to five groups: normal control,negative control(Lipo2000+ pGenesil-2.1 HK),Lipo2000+pGenesil-2.1-PTTG siRNA,Lipo2000+ pGenesil-2.1-survivin siRNA and Lipo2000+pGenesil-2.1-PTTG-survivin siRNA groups.The PTTG and survivin protein and mRNA levels in every group were measured with flow cytometry,Western blotting and RT-PCR method after U251 cells were transfected with these plasmids.Results The PTTG and survivin protein and mRNA levels in Lipo2000+pGenesil-2.1 PTTG siRNA,Lipo2000+ pGenesil-2.1 survivin siRNA and Lipo2000+pGenesil-2.1-PTTG-survivin siRNA groups were decreased significantly compared with negative control group(P0.01),especially in pGenesil-2.1-PTTG-survivin siRNA group.Conclusion The silence effects of double interfering vector pGenesil-2.1-PTTG-survivin siRNA on PTTG and survivin gene in U251 cells are stronger than those of pGenesil-2.1-PTTG siRNA or pGenesil-2.1-survivin siRNA.
出处 《吉林大学学报(医学版)》 CAS CSCD 北大核心 2012年第1期37-41,共5页 Journal of Jilin University:Medicine Edition
基金 教育部博士点基金资助课题(20070183193) 吉林省科技厅自然科学基金资助课题(200705108 200505228)
关键词 垂体肿瘤转化基因 生存素 RNA干扰 胶质瘤 pituitary tumor transforming gene survivin RNAi glioma
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同被引文献16

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