| siRNA抑制缺氧人视网膜色素上皮细胞ILK的表达及对HIF-1α的影响 |
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| 引用本文： | 张奕霞,曾爱萍,曾水清.siRNA抑制缺氧人视网膜色素上皮细胞ILK的表达及对HIF-1α的影响[J].国际眼科杂志,2008,8(4):708-710. |
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| 作者姓名： | 张奕霞 曾爱萍 曾水清 |
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| 作者单位： | 1. 430022,中国湖北省武汉市,华中科技大学同济医学院附属协和医院眼科;832002,新疆维吾尔自治区石河子市,石河子大学医学院一附院眼科
2. 华中科技大学同济医学院附属协和医院眼科,中国湖北省武汉市,430022 |
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| 摘 要： | 目的:探讨干扰RNA(siRNA)沉默缺氧培养下人的视网膜色素上皮细胞(hRPE)中整合素连接激酶(ILK)的表达及其对缺氧诱导因子1α(HIF-1α)表达的影响。方法:CoCl2建立hRPE细胞的化学缺氧模型,Western blot和RT-PCR方法半定量检测不同缺氧时间(0,6,12,24,48h)hRPE细胞中ILK、HIF-1α蛋白及其mRNA的表达;阳离子脂质体转染ILK的干扰片段抑制缺氧24h hRPE细胞中ILK的表达,同上方法检测转染后缺氧24h hRPE细胞中ILK的表达及其对HIF-1α表达的影响。结果:ILK表达于正常及缺氧培养的hRPE细胞中。随着hRPE细胞缺氧时间的延长,在蛋白和mRNA水平ILK、HIF-1α都呈现逐渐增加的表达趋势。siRNA抑制ILK在缺氧24h hRPE细胞中的表达,同时显著抑制了HIF-1α的表达,较阴性对照组、单纯脂质体组有显著差异(P<0.01)。结论:ILK可以通过HIF途径应答RPE细胞的缺氧反应。
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| 关 键 词： | 缺氧 RNA干扰 ILK HIF-1α |
| 修稿时间： | 2008年1月13日 |
Effect of integrin-linked kinase siRNA-mediated gene silencing on hypoxia human retinal pigment epithelial cell and the expression of HIF-1α |
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| Yi-Xia Zhang,Ai-Ping Zeng,Shui-Qing Zeng.Effect of integrin-linked kinase siRNA-mediated gene silencing on hypoxia human retinal pigment epithelial cell and the expression of HIF-1α[J].International Journal of Ophthalmology,2008,8(4):708-710. |
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| Authors: | Yi-Xia Zhang Ai-Ping Zeng Shui-Qing Zeng |
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| Institution: | 1Department of Ophthalmology,the Affiliated Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,Hubei Province,China;2Department of Ophthalmology,the First Affiliated Hospital,School of Medicine,Shihezi University,Shihezi 832002,Xinjiang Uygur Autonomous Region,China |
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| Abstract: | AIM:To study the expression of integrin-linked kinase(ILK)in hypoxic human retinal pigment epithelial(hRPE)cell with siRNA-mediated gene silencing and the influence on the expression of HIF-1α.METHODS:COCl2(150 μmol/L)was used to establish the chemical hypoxic model in hRPE cell.Western blot and RT-PCR were used to semiquantitatively evaluate the expressions of ILK and HIF-1α in protein and their mRNA levels at different hypoxia time(0,6,12,24 and 48 hours).ILK intervention fragment transfected with liposome 2000 was inserted into hypoxia hRPE cells,and the expression of ILK was silenced.The expression of ILK and the influence on HIF-1α were detected with Western blot and RT-PCR after transfection.RESULTS:ILK was found in normal and hypoxia hRPE.The expressions of ILK and HIF-1α increased with time extension on hypoxia.SiRNA transfection significantly depressed the levels of ILK and HIF-1α.CONCLUSION:ILK could respond to hypoxia via HIF pathway in hRPE. |
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| Keywords: | hypoxia siRNA integrin-linked kinase HIF-1α |
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