山东大学耳鼻喉眼学报 ›› 2016, Vol. 30 ›› Issue (5): 110-114.doi: 10.6040/j.issn.1673-3770.0.2016.322

• 论著 • 上一篇    下一篇

RNAi沉默PDCD4基因对人喉癌Hep-2细胞增殖及β-catenin表达的影响

许元腾1,陈瑞庆2,林功标1,方秀玲1,俞舒娟1,梁晓华3,张榕1   

  1. 福建医科大学附属第一医院 1.耳鼻咽喉科;
    2.中心实验室;
    3.检验科, 福建 福州 350005
  • 收稿日期:2016-07-20 出版日期:2016-10-20 发布日期:2016-10-20
  • 通讯作者: 林功标. E-mail: lingongbiao@qq.com; 张榕. E-mail:zhang2012rong@sina.cn E-mail:xyt973@163.co
  • 作者简介:许元腾. E-mail:xyt973@163.co
  • 基金资助:
    福建省自然科学基金(2014j01311);福建省财政厅专项基金［闽财指(2015)1297号］

Effect of silencing PDCD4 gene on proliferation of Hep-2 cells and the expression of β-catenin by RNA interference technique.

XU Yuanteng1, CHEN Ruiqing2, LIN Gongbiao1, FANG Xiuling1, YU Shujuan1, LIANG Xiaohua3, ZHANG Rong   

  1. 1. Department of Otolaryngology;2. Central Laboratory;3. Clinical Laboratory, The First Affliated Hospital of Fujian Medical University, Fuzhou 350005, Fujian, China
  • Received:2016-07-20 Online:2016-10-20 Published:2016-10-20

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摘要： 目的 应用RNAi技术下调人喉癌Hep-2细胞中PDCD4基因的表达,探讨其对Hep-2细胞增殖及β-catenin表达的影响。 方法 设计并合成针对PDCD4基因的shRNA质粒与阴性对照质粒,分别转染Hep-2细胞。实时定量RT-PCR和Western blotting检测转染前后阴性对照组与干扰组PDCD4、β-catenin基因mRNA和蛋白的表达。克隆形成实验观察Hep-2细胞增殖能力的变化。 结果 与对照组相比,PDCD4干扰组的Hep-2细胞克隆形成率显著升高(P=0.01),PDCD4干扰组PDCD4mRNA和蛋白均显著减低(P<0.01)、β-catenin mRNA和蛋白均较对照组显著增加(P<0.01)。 结论 成功应用RNAi技术下调人喉癌Hep-2细胞中PDCD4基因的表达,Hep-2细胞克隆形成能力增强,β-catenin mRNA和蛋白表达显著升高。

关键词: RNA干扰, 喉癌, Hep-2细胞, β-catenin, 细胞增殖, PDCD4基因

Abstract: Objective To study the effect of silencing PDCD4 gene on proliferation of Hep-2 cells and the expression of β-catenin by RNA interference technique. Methods Specific shRNA plasmid and negative control plasmid of PDCD4 gene were designed and synthesized. Hep-2 cells were transfected with shRNA plasmid and negative control plasmid. Real-time RT-PCR and Western blotting analysis were performed to determine the expressions of PDCD4 and β-catenin mRNA and protein after transfection, respectively. The proliferation of Hep-2 cells was observed by colony-forming assay. Results Compared to the control group, the colony formation rate of Hep-2 cells was significantly increased in the shRNNA plasmid group(P=0.01); the expressions of PDCD4 mRNA and protein were significantly reduced(P<0.01);the expressions of β-catenin mRNA and protein were significantly increased(P<0.01). Conclusion RNA interference technique can effectively silence PDCD4 gene expression of Hep-2 cells,enhance the colony forming ability of Hep-2 and increase the expressions of β-catenin mRNA and protein.

Key words: Hep-2 cell, PDCD4 gene, β-catenin, Cell proliferation, Laryngeal cancer, RNA interference

中图分类号: 

  • R739.65
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