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)