Silence the expression of c-erbB-2 and Bcl-2 gene in Tca8113 cell line by rnai technique

JI Ping; et al

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Silence the expression of c-erbB-2 and Bcl-2 gene in Tca8113 cell line by rnai technique

更新时间：2024-01-17

- Silence the expression of c-erbB-2 and Bcl-2 gene in Tca8113 cell line by rnai technique
- Journal of Chongqing Medical University Issue 2, Pages: 163-168(2006)
- 作者机构：
  
  1. 重庆医科大学附属口腔医院颌面外科
  2. 重庆医科大学附属口腔医院颌面外科,重庆,400016
- 作者简介：
- 基金信息：
- DOI：
  CLC： Q78
- Published：2006，
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[1]季平,张福军,李少林.表达重组干扰质粒对Tca8113细胞c-erbB-2、Bcl-2基因表达干扰作用的实验研究[J].重庆医科大学学报,2006(02):163-168+179.

JI Ping, et al. Silence the expression of c-erbB-2 and Bcl-2 gene in Tca8113 cell line by rnai technique. [J]. Journal of Chongqing Medical University (2):163-168(2006)
[1]季平,张福军,李少林.表达重组干扰质粒对Tca8113细胞c-erbB-2、Bcl-2基因表达干扰作用的实验研究[J].重庆医科大学学报,2006(02):163-168+179. DOI：

JI Ping, et al. Silence the expression of c-erbB-2 and Bcl-2 gene in Tca8113 cell line by rnai technique. [J]. Journal of Chongqing Medical University (2):163-168(2006) DOI：

摘要

目的:探讨表达重组干扰质粒（RNAi技术）对人舌鳞癌细胞株（Tca8113）细胞中c-erbB-2、Bcl-2基因表达的干扰效率。方法:按siRNA设计原则设计针对c-erbB-2、Bcl-2基因的OligoDNA

体外化学合成Oligo DNA

Oligo DNA退火、连接、转化、筛选克隆

构建针对c-erbB-2基因的表达重组RNA干扰质粒（psiC1、psiC2、psiC3）和针对Bcl-2基因的表达重组RNA干扰质粒（psiB1、psiB2、psiB3）

细胞脂质体转染Tca8113细胞

Real-time RT-PCR的标准曲线、扩增曲线和熔解曲线的数据收集处理

采用荧光定量RT-PCR方法检测重组干扰质粒对Tca8113细胞中c-erbB-2、Bcl-2基因表达的干扰效率。结果:实时荧光定量RT-PCR方法检测psiC1、siC2、psiC3质粒脂质体转染后c-erbB-2基因的表达情况结果显示psiC2、psiC3相对于对照组均有不同程度的干扰效果

psiC1相对于对照没有干扰效果。psiC2的干扰程度较大为62.68%

psiC3的干扰程度较小为40.61%。psiB1、psiB3相对于对照组无明显干扰效果

psiB2的干扰程度高达66.20%。结论:针对c-erbB-2、Bcl-2基因的表达重组RNA干扰质粒psiC2（gAgTCCCAACCATgTCAAA）、psiB2（CCgggAgATAgTgATgAA）能有效沉默Tca8113细胞中c-erbB-2、Bcl-2基因的表达。

Abstract

Objective: To investigate the interfering efficiency of RNAi technique on the expression of c-erbB-2 and Bcl-2 oncogene in human tongue squamous carcinoma(Tca8113).Methods: The recombined RNAi plasmids for c-erbB-2 and Bcl-2 oncogene were constituted by the four succussive steps-designing of Oligo DNAs

synthesis of Oligo DNAs

transfection of Oligo DNAs into pSUPER.neo+gfp vectors and selection of positive plasmids.In order to silence the expression of c-erbB-2 and Bcl-2 oncogenes

the recombined RNAi plasmids were transfected into Tca8113 cells by culturing together for about 10 hours

and the interfering efficiency of RNAi for the two oncogenes was evaluated by fluorescence-quantitative RT-PCR.Results: The interfering efficiencies for c-erbB-2 oncogene were 62.68% and 40.61%

respectively in psiC1?psiC2?psiC3

while the interfering efficiencies for Bcl-2 oncogene were 66.20% and respectively in psiB1?siB2?psiB3.Conclusion: The recombined RNAi plasmids of psiC2(gAgTCCCAACCATgTCAAA) for c-erbB-2 oncogene and of psiB2(CCgggAgATAgTgATgAA) for Bcl-2 oncogene can effectively silence the expression of c-erbB-2 and Bcl-2 oncogenes in Tca8113 cell.

关键词

重组干扰质粒脂质体转染基因表达实时荧光定量RT-PCR

Keywords

Recombined interfering plasmidLiposomes transfectiongene expressionFluorescence-quantitative RT-PCR

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