siRNA真核表达载体的构建及其对CXCR4基因的沉默效应
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唐利立

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Construction of eukaryotic expression vector of siRNA and its silence effects on CXCR4 gene
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    摘要:

    目的:构建趋化因子受体(CXCR4) 特异性siRNA ( small interfere RNA)真核表达载体,体外观察对CXCR4基因的沉默作用。方法:采用基因克隆技术, 将合成的特异性CXCR4 RNA干扰寡核苷酸序列插入真核表达载体pSUPER, 构建CXCR4 siRNA真核表达载体。以脂质体法将pSUPER空载体和2个(pSUPER/CXCR4siRNA0和pSUPER/ CXCR4siRNA1 )重组质粒分别导入具有高转移特性的MDAMB231乳腺癌细胞系,检测各组乳腺癌细胞内CXCR4 mRNA及蛋白的表达情况。 结果:成功构建CXCR4 siRNA真核表达载体. 转染CXCR4 RNA干扰片段的乳腺癌细胞, 72 h后CXCR4 mRNA及蛋白表达下调,抑制作用明显。 结论:构建的RNA干扰真核表达载体能明显下调CXCR4 mRNA及蛋白的表达,为应用于乳腺癌的治疗研究提供了一定的实验基础。

    Abstract:

    Abstract:Objective:To construct the small interfering RNA (siRNA) eukaryotic expression vector specific for human CXCR4 gene and to observe its silencing effects on CXCR4 gene. Methods:The expression vectors of pSUPER/ CXCR4siRNA1 and the control group pSUPER/ CXCR4siRNA0 were constructed by gene recombination, and then were transfected by liposome mediated into the MDAMB231 breast cancer cells with high metastasis potential . The expression of CXCR4 mRNA and protein was detected. Results:The eukaryotic expression vectors of pSUPER/ CXCR4siRNA1 ,which downregulated mRNA and protein of CXCR4 at 72 h after transfection, were successfully constructed. The inhibitory effects of pSUPER/CXCR4 siRNA1 was obvious stastictically. Conclusions:Eukaryotic expression vector of siRNA specific for CXCR4 is successfully constructed and has inhibitory effects obviously, which can provide the experimental basis for its application in the treatment of breast cancer.

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何英,唐利立. siRNA真核表达载体的构建及其对CXCR4基因的沉默效应[J].中国普通外科杂志,2006,15(10):4-740.
DOI:10.7659/j. issn.1005-6947.2006.10.004

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  • 在线发布日期: 2006-10-25