靶向PTTG和survivin基因共干扰质粒对U251细胞的沉默效率

J4 ›› 2012, Vol. 38 ›› Issue (1): 37-41.

靶向PTTG和survivin基因共干扰质粒对U251细胞的沉默效率

徐松柏¹|刘宝斌¹|赵红光²|于洪泉¹|李蕴潜¹|张显峰¹|张宇¹|金铮¹|赵刚¹

（1. 吉林大学第一医院神经外科|吉林长春 130021；2. 吉林大学第一医院核医学科|吉林长春 130021)

收稿日期:2011-04-02 出版日期:2012-01-28 发布日期:2012-01-28
通讯作者: 赵刚 E-mail:(Tel:0431-88782463,E-mail：jackieshew@sina.com)
作者简介:徐松柏（1978-）|男|吉林省长春市人|讲师|医学博士|主要从事胶质瘤基因治疗的研究。
基金资助:
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）

Silence effects of double interfering vector targeting PTTG and survivin genes on U251 cells

XU Song-bai¹,LIU Bao-bin¹,ZHAO Hong-guang²,YU Hong-quan¹,LI Yun-qian¹,ZHANG Xian-feng¹,ZHANG Yu,JIN Zheng¹|ZHAO Gang¹

(1. Department of Neurosurgery,First Hospital,Jilin University,Changchun 130021,China; 2. Department of Nuclear Medicine,First Hospital,Jilin University,Changchun 130021,China)

Received:2011-04-02 Online:2012-01-28 Published:2012-01-28

摘要/Abstract

摘要：

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+ 〖JP〗pGenesil-2.1 HK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Western blotting方法检测转染36 h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。

关键词: 垂体肿瘤转化基因；生存素；RNA干扰；胶质瘤

Abstract:

Abstract: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（P<0.01）,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.

Key words: pituitary tumor transforming gene;survivin;RNAi;glioma

中图分类号:

徐松柏|刘宝斌|赵红光|于洪泉|李蕴潜|张显峰|张宇|金铮|赵刚. 靶向PTTG和survivin基因共干扰质粒对U251细胞的沉默效率[J]. J4, 2012, 38(1): 37-41.

XU Song-bai,LIU Bao-bin,ZHAO Hong-guang,YU Hong-quan,LI Yun-qian,ZHANG Xian-fen. Silence effects of double interfering vector targeting PTTG and survivin genes on U251 cells[J]. J4, 2012, 38(1): 37-41.