山东大学耳鼻喉眼学报 ›› 2015, Vol. 29 ›› Issue (5): 38-42.doi: 10.6040/j.issn.1673-3770.0.2015.143

• 论著 • 上一篇    下一篇

沉默HIF-1α基因对鼻黏膜上皮细胞生长因子表达的影响

李盈盈, 周涵, 张伟强, 董伟达   

  1. 南京医科大学第一附属医院, 江苏省人民医院耳鼻咽喉科, 江苏 南京 210029
  • 收稿日期:2015-04-07 修回日期:2015-07-09 出版日期:2015-10-16 发布日期:2015-10-16
  • 通讯作者: 董伟达,E-mail:weidadong2649@126.com;周涵,E-mail:zhouhan1979@sina.com E-mail:weidadong2649@126.com;zhouhan1979@sina.com
  • 作者简介:李盈盈,E-mail:liyingyer@yeah.net
  • 基金资助:
    国家自然科学基金(81200729)

Effects of silencing hypoxia inducible factor-1α on the expression of cytokines in cultured human nasal epithelial cells

LI Yingying, ZHOU Han, ZHANG Weiqiang, DONG Weida   

  1. Department of Otorhinolaryngology, The First Affiliated Hospital with Nanjing Medical University, Jiangsu Provence Hospital, Nanjing 210029, Jiangsu, China
  • Received:2015-04-07 Revised:2015-07-09 Online:2015-10-16 Published:2015-10-16

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摘要： 目的 采用短发夹 RNA(shRNA)特异性沉默人鼻黏膜上皮细胞(HNEC)缺氧诱导因子-1α (HIF-1α)基因,观察对其表达血管内皮生长因子(VEGF)、转化生长因子(TGF)β1、碱性成纤维细胞生长因子(bFGF) 的影响。方法 体外培养原代鼻黏膜细胞,设计合成以腺病毒为载体GFP标记的shRNA,将HNEC分为空白对照组、阴性对照组(ad-HIF shRNA-neg)、ad-HIF shRNA干扰组。采用RT-PCR和Western blotting技术检测转染后相应因子蛋白和mRNA的表达情况。结果 成功合成ad-HIF shRNA并转染入HNEC中。转染ad-HIF shRNA后,HIF-1α蛋白及mRNA表达分别下降40%和39%(q=31.469,16.590);同时VEGF、TGF-β1、bFGF的蛋白及mRNA表达也随之下降,差异均有统计学意义(P<0.05)。结论 HIF-1α mRNA的shRNA能有效地使HNEC的HIF-1α基因沉默,进而有效地抑制VEGF、TGF-β1和bFGF的表达, 提示HNEC中HIF-1α可能通过直接或间接途径调控着VEGF、bFGF和TGF-β1的表达。

关键词: 鼻黏膜, 血管内皮生长因子, 碱性成纤维细胞生长因子, 缺氧诱导因子-1, 转化生长因子β1, RNA干扰

Abstract: Objective To investigate the effects of silencing hypoxia inducible factor-1α(HIF-1α) by transfecting short hairpin RNA(shRNA)into cultured human nasal epithelial cells (HNECs) and its further regulations on vascular endothelial growth factor(VEGF), transforming growth factor(TGF)β1 and basic fibroblast growth factor(bFGF). Methods HNECs were isolated from 10 inferior turbinate-cutting patients'mucosa. Two HIF-shRNA fragments were built and the best silencing shRNA sequence was used for transfection. We separated cells into three groups: the blank group, the negative control group and the ad-HIFshRNA interference group. The protein and gene expressions were detected by Western blotting and PCR. Results HNECs were transfected with GFP marketed adenovirus which carry the best silencing HIF-1α shRNA. The protein and mRNA expressions of HIF-1α were significantly reduced 40% and 39%(q=31.469, 16.590), and following, the protein and mRNA expressions of VEGF, TGF-β1 and bFGF were reduced (P<0.05). Conclusion Transfection of ad-HIFshRNA can effectively silence the expression of HIF-1α gene and protein and furthermore inhibit the expressions of VEGF, TGF-β1 and FGF-2, which suggests HIF-1α regulates the expression of VEGF, bFGF and TGF-β1 through direct or indirect ways. Thus silencing HIF-1α may provide a new way towards allergic rhiniti's treatment.

Key words: Nasal epithilial cells, RNA interference, Hypoxia-inducible factor-1α, Transforming growth factor-β1, Vascular endothelial growth factor, Basic fibroblast growth factor

中图分类号: 

  • R765.2
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