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山东大学学报（医学版） ›› 2014, Vol. 52 ›› Issue (11): 11-15.doi: 10.6040/j.issn.1671-7554.0.2014.263

• 基础医学 • 上一篇    下一篇

靶向EZH2的shRNA表达对SW480细胞的抑制作用及5-Fu对其影响的研究

吕艳锋1,2, 姜华2, 王建新2, 张成博3   

  1. 1. 山东中医药大学, 山东 济南 250014;
    2. 山东大学第二医院肛肠外科, 山东 济南 250033;
    3. 山东中医药大学文献研究所, 山东 济南 250014
  • 收稿日期:2014-04-27 发布日期:2014-11-10
  • 通讯作者: 张成博.E-mail:youtow@sina.com E-mail:youtow@sina.com
  • 基金资助:
    山东省自然科学基金(ZR2011HQ049);济南市科技明星计划(201004055,20090208)

Inhibitive effect of EZH2-specific shRNA on SW480 cells and the impact of 5-FU

LV Yanfeng1,2, JIANG Hua2, WANG Jianxin2, ZHANG Chengbo3   

  1. 1. Shandong University of Traditional Chinese Medicine, Jinan 250014, Shandong, China;
    2. Department of Anoproctology, Second Hospital of Shandong University, Jinan 250033, Shandong, China;
    3. Literature Research Institute, Shangdong University of Traditional Chinese Medicine, Jinan 250014, Shandong, China
  • Received:2014-04-27 Published:2014-11-10

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摘要： 目的 研究EZH2特异性shRNA对人结肠癌细胞株SW480细胞的抑制作用和化疗药物5-FU对其干预作用的影响.方法 体外将EZH2特异性shRNA表达载体转染SW480细胞,RT-PCR和Western blotting检测细胞EZH2 mRNA及蛋白的表达,MTT法检测细胞增殖活性;将靶向EZH2的shRNA表达的SW480细胞接种于裸鼠,5-FU对其进行干预,RT-PCR检测瘤组织EZH2 mRNA的表达.结果 与阴性对照组相比,基因沉默组EZH2 mRNA表达明显降低(P<0.01),EZH2蛋白表达明显降低(P<0.05);与空白对照组相比,基因沉默组细胞生长受到明显抑制(P<0.05);接种于裸鼠后,与正常组相比,5-FU基因沉默组与基因沉默组瘤组织体积、质量、EZH2 mRNA的表达均明显降低(P<0.01),与基因沉默组相比,5-FU基因沉默组EZH2 mRNA的表达明显降低(P<0.05).结论 靶向EZH2的shRNA表达在体外及体内均可显著抑制SW480细胞EZH2 mRNA 和蛋白表达及细胞生长;5-FU能显著抑制体内EZH2基因沉默的SW480细胞EZH2 mRNA的表达.

关键词: EZH2基因, RNA干扰, SW480细胞, 5-Fu

Abstract: Objective To investigate the inhibitive effect of EZH2-specific shRNA on human colon cancer cell line SW480 and the impact of 5-FU on this inhibitive effect. Methods SW480 cells were transfected with EZH2-specific shRNA in vitro, then EZH2 mRNA and protein expressions were detected with RT-PCR and Western blotting, and cell proliferation was examined with MTT. The EZH2-specific shRNA expressing SW480 cells were implanted to nude mice and treated with 5-FU. After that, the levels of EZH2 mRNA expressed by tumor tissues were measured with RT-PCR. Results Compared with that of the negative control group, the EZH2 mRNA expression (P<0.01) and protein expression (P<0.05) of the gene-silencing group markedly decreased. The cell growth of the gene-silencing group was inhibited compared with that of the control group (P<0.05). In nude mice, the tumor volume, tumor weight and EZH2 mRNA expression of the 5-FU gene-silencing group and of the gene-silencing group remarkably decreased compared with those of the normal group (P<0.01). The EZH2 mRNA expression of the 5-FU gene-silencing group significantly decreased compared with that of the gene-silencing group (P<0.05). Conclusion The EZH2-specific shRNAplays a significant inhibitive role on EZH2 mRNA and protein expressions in SW480 cells both in vitro and in vivo. 5-FU can significantly inhibit the EZH2 mRNA expression of EZH2-gene-silencing SW480 cells in vivo.

Key words: EZH2 gene, SW480 cells, RNA interference, 5-Fu

中图分类号: 

  • R735.3
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