shRNA-mediated interference of gene expression PLCε downturn on T24 bladder cancer cells

LUO Chun-li; et al

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shRNA-mediated interference of gene expression PLCε downturn on T24 bladder cancer cells

更新时间：2024-01-17

- shRNA-mediated interference of gene expression PLCε downturn on T24 bladder cancer cells
- Journal of Chongqing Medical University Vol. 34, Issue 10, Pages: 1313-1316(2009)
- 作者机构：
  
  1. 重庆医科大学检验系教育部重点实验室
  2. 重庆医科大学附属第一医院泌尿外科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R737.14
- Published：2009，
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[1]罗春丽,郭永灿,赵懿,欧俐苹,颜令,朴军,吴小候.shRNA干扰介导PLCε基因表达下调对膀胱癌T24细胞生长的影响[J].重庆医科大学学报,2009,34(10):1313-1316.

LUO Chun-li, et al. shRNA-mediated interference of gene expression PLCε downturn on T24 bladder cancer cells. [J]. Journal of Chongqing Medical University 34(10):1313-1316(2009)
[1]罗春丽,郭永灿,赵懿,欧俐苹,颜令,朴军,吴小候.shRNA干扰介导PLCε基因表达下调对膀胱癌T24细胞生长的影响[J].重庆医科大学学报,2009,34(10):1313-1316. DOI：

LUO Chun-li, et al. shRNA-mediated interference of gene expression PLCε downturn on T24 bladder cancer cells. [J]. Journal of Chongqing Medical University 34(10):1313-1316(2009) DOI：

摘要

目的:探讨shRNA干扰沉默PLCε基因对人膀胱癌T24细胞生长的影响。方法:体外构建PLCε基因的shRNA的重组质粒

Lipofectamine2000介导法转染T24细胞

采用RT-PCR方法检测特异性shRNA的重组质粒对PLCε基因沉默效果

转染后采用MTT法检测shRNA的重组质粒对细胞增殖的作用

免疫细胞化学染色法检测T24细胞PCNA的表达

电镜观察细胞超微结构的改变。结果:shRNA的重组质粒能有效下调PLCε基因的表达

抑制率为78.01%

与对照组比较其差异有统计学意义（P<0.01）;细胞增殖活性受到明显抑制

与对照组比较其差异有统计学意义（P<0.01）;细胞内PCNA含量明显低于空白对照组和阴性质粒组

其差异有统计学意义（P<0.01）;细胞形态改变产生凋亡小体。结论:PLCε基因有望成为应用RNAi技术探索治疗膀胱癌潜在的靶基因。

Abstract

Objective:To investigate whether PLCε gene down regulation by RNA interference(RNAi) leads to inhibition of proliferation in human bladder carcinoma T24 cell.Methods: The shRNA recombinant plasmids targeting to PLCε gene was constructed and transfected into bladder carcinoma T24 cell with Lipofectamine 2000.RT-PCR was used to monitor the validity of specific s h R N A in down regulation of PLCε.Then MTT assay was performed for detecting cell proliferation

the changes of PCNA were analyzed by immunocytochemical method

Electron microscope was used to observe the morphological changes.Results: The specific PLCε shRNA was confirmed to be efficient in silencing PLCε expression.PLCε gene down regulation by the shRNA recombinant plasmids inhibition cell proliferation rate about 78.01%

They were significantly different from that of control group(P<0.01;After transfected the specific recombinant plasmids

PCNA expression was significantly decreased

They were significantly different from that of control group(P<0.01;morphological changes produce apoptotic body.Conclusion: PLCεis likely to be potential molecular target for bladder carcinoma in gene therapy by RNAi.

关键词

膀胱癌PLCε基因RNA干扰细胞增殖增殖细胞核抗原

Keywords

Bladder carcinomaPLCεG geneRNA interferencecell proliferationproliferating cell nuclear antigen

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