【摘要】 目的构建趋化因子受体(CXCR 4)特异性siRNA(small interfere RNA)真核表达载体,体外观察对CXCR 4基因的沉默作用。方法采用基因克隆技术,将合成的特异性CXCR 4 RNA干扰寡核苷酸序列插入真核表达载体pSUPER,构建CXCR 4 siRNA真核表达载体。以脂质体法将pSUPER空载体和2个(pSUPER/CXCR 4-siRNA 0和pSUPER/CXCR 4-siRNA 1)重组质粒分别导入具有高转移特性的MDA-MB-2 3 1乳腺癌细胞系,检测各组乳腺癌细胞内CXCR 4 mRNA及蛋白的表达情况。结果成功构建CXCR 4 siRNA真核表达载体.转染CXCR 4 RNA干扰片段的乳腺癌细胞,7 2 h后CXCR 4 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CXCR 4 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。更多还原

【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.更多还原