骨桥蛋白_____骨桥蛋白；小干扰RNA；真核表达

Osteopontin特异性siRNA真核表达载体的构建及其在GC9811胃癌细胞中沉默效应的鉴定

http://www.100md.com 《第四军医大学学报》 2005年第3期

骨桥蛋白,,骨桥蛋白；小干扰RNA；真核表达,0引言,1材料和方法,2结果,3讨论

     Construction of eukaryotic expression vector of siRNA specific for osteopontin and characterization of its efficiency in GC9811 gastric carcinoma cell line

    TANG HongWei1,2, PAN YangLin1, NIE YongZhan1, MENG FanPing1, XIE HuaHong1, WANG Jun1, WU KaiChun1

    1Institute of Gastrointestinal Diseases Research, Xijing Hospital, Fourth Military Medical University, Xian 710033, 2Department of Gastroenterology, General Hospital, Chinese Peoples Armed Police Forces, Beijing 100039, China

    【Abstract】 AIM: To construct the small interfering RNA (siRNA) eukaryotic expression vector specific for human OPN gene and to observe its silencing effect on OPN gene. METHODS: The expression vectors of mU6pro/OPNsiRNA1 and mU6pro/OPNsiRNA2 were constructed by gene recombination and then were transfected by liposomemediated transfection into the GC9811 gastric carcinoma cell line with high metastasis potential recently established in our laboratory. At 72 h after transfection, the expression of OPN in the levels of mRNA and protein was detected by RTPCR and Westernblotting. RESULTS: The eukaryotic expression vectors of mU6pro/OPNsiRNA1 and mU6pro/OPNsiRNA2, which downregulated mRNA and protein of OPN at 72 h after transfection, were successfully constructed. mU6pro/OPNsiRNA1 was more effective than the other one. CONCLUSION: Eukaryotic expression vector of siRNA specific for OPN is successfully constructed, which lays the basis for its application in the treatment of gastric carcinoma. ......

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