Inhibition of survivin gene expression in EJ cells by RNAi

YANG Hua-an; et al

您当前的位置：

首页 >

文章列表页 >

Inhibition of survivin gene expression in EJ cells by RNAi

更新时间：2024-01-17

- Inhibition of survivin gene expression in EJ cells by RNAi
- Journal of Chongqing Medical University Issue 1, Pages: 33-36(2008)
- 作者机构：
  
  1. 重庆医科大学附属第一医院泌尿外科
  2. 重庆医科大学附属第一医院泌尿外科,重庆,400016
- 作者简介：
- 基金信息：
- DOI：
  CLC： R737.14
- Published：2008，
扫描看全文
[1]杨华安,苟欣,吴跃,郑传东.RNA干扰技术阻断EJ细胞中survivin基因的表达[J].重庆医科大学学报,2008(01):33-36.

YANG Hua-an, et al. Inhibition of survivin gene expression in EJ cells by RNAi. [J]. Journal of Chongqing Medical University (1):33-36(2008)
[1]杨华安,苟欣,吴跃,郑传东.RNA干扰技术阻断EJ细胞中survivin基因的表达[J].重庆医科大学学报,2008(01):33-36. DOI：

YANG Hua-an, et al. Inhibition of survivin gene expression in EJ cells by RNAi. [J]. Journal of Chongqing Medical University (1):33-36(2008) DOI：

摘要

目的:用真核转录载体pSilencer2.0构建针对survivin基因的重组真核转录载体

并转染膀胱肿瘤细胞系EJ

通过RNA干扰（RNAi）阻断EJ细胞中survivin基因的表达。方法:将合成的DNA正义链及反义链变性、退火

形成的双链DNA与pSilencer2.0线性质粒以T4DNA连接酶连接

重组质粒经酶切、DNA测序鉴定后

用脂质体法转染EJ细胞

通过RT-PCR、免疫印迹检测干扰效果。结果:酶切及测序证实设计合成的DNA已正确插入载体。RT-PCR、免疫印迹检测实验表明:pSilencer2.0-SVV1、pSilencer2.0-SVV2重组载体有效地阻断了EJ细胞中survivin基因的表达

survivin基因在mRNA和蛋白水平上的表达明显下调（P<0.01）

其中pSilencer2.0-SVV2重组载体干扰效果更佳。结论:成功构建了针对survivin基因的真核转录载体pSilencer2.0-SVV1、pSilencer2.0-SVV2

两者均可有效地阻断膀胱癌细胞系EJ细胞中survivin基因的表达

pSilencer2.0-SVV2重组载体效果更佳

这为后继实验研究打下基础。

Abstract

Objective:To construct eukaryotic expression vectors with the pSilencer2.0 vector for inhibiting human survivin gene by RNA interference

and to detect the effect of the silenced survivin gene on EJ cells.Methods:Two target gene segments were synthesized and cloned into the pSilencer2.0 vector respectively to construct two recombinant eukaryotic expression vectors

pSilencer2.0-SVV1and pSilencer2.0-SVV2

which were identified by enzyme digestion analysis and DNA sequencing.Then the EJ cells were transfected with the recombinant vectors by lipofection and the interference effect was detected by RT-PCR and Western-blot.Results:Enzyme digestion analysis and DNA sequencing showed that two target segments were cloned into pSilencer2.0 vectors correctly.The results of RT-PCR and Western blot indicated that pSilencer2.0-SVV1 and pSilencer2.0-SVV2 vectors could significantly knock down the transcription and expression of survivin gene(P<0.01)

and the latter was better.Conclusion:pSilencer2.0-SVV1 and pSilencer2.0-SVV2 vectors for survivin gene were constructed successfully.The transcription and expression of survivin gene were inhibited effectively by the constructed vectors in the bladder cancer cell line EJ and the latter was better.This paves way for the subsequent studies.

关键词

RNA干扰Survivin膀胱癌细胞

Keywords

RNA interferenceSurvivinBladder cancer cell

references

Views

113

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Molecular mechanism of Buzhong Yiqi Decoction in coordination with Siβ-catenin in intervention against cisplatin resistance in lung adenocarcinoma by regulating the Wnt/β-catenin signaling pathway

RNAi technology applied to the preparation of molecular probes for diagnosis and treatment of ovarian cancer

Construction of recombinant lentiviral vector of si RNA for transforming growth factor-beta typeⅠ receptor gene and its expression in human embryonic lung fibroblasts

Effect of GM130 on O-linked glycosylation and adhesion of human gastric cancer cell line MKN-45

Activation of Wnt/β-catenin signaling pathway promotes anti-apoptotic genes bcl-2 and survivin expression in H9c2 cell subject to chronic hypoxia

Related Author

Li He

Mu Qirui

Wang Zhe

Wang Ying

Yu Dan

Gao Yuan

Wu Xiaofeng

Wen Ming

Related Institution

Department of Pathology，College of Integrative Medicine，Liaoning University of Traditional Chinese Medicine

Department of Radiology,The First Affiliated Hospital of Chongqing Medical University

Department of Emergency and Intensive Care Unit,The First Affiliated Hospital of Chongqing Medical University

Teaching and Research Section of Pathology,College of Basic Medicine,Molecular Medicine and Cancer Research Center,Chongqing Medical University

Department of Cardiology,Children’s Hospital of Chongqing Medical University

⁰