中文摘要：

目的在体外研究中证明神经胶质纤维酸性蛋白（GFAP）启动子能引导人钠碘转运体基因（hNIS）实现胶质瘤靶向性表达，并能有效介导放射性碘治疗胶质瘤。方法使用Lipofectamine2000脂质体将PGL3．Basic、PGL3-Control、PGL3．GFAP质粒分别转染至U251、U87、MRC-5细胞，24h后在化学发光仪中检测各细胞的相对反应活性；构建GFAP启动子引导hNIS基因表达的重组腺病毒Ad．GFAP．hNIS，转染至各细胞后Westernblotting检测细胞GFAP、hNIS蛋白的表达；同时设Ad—CMV—EGFP组（空白对照）和Ad．CMV—hNIS组（阳性对照），应用y计数器测量3组细胞的131I摄碘和外流能力，龙胆紫染色后记数细胞并计算克隆形成率：U87细胞接种BALB／c雌性裸鼠20只，按随机数字表法分4组（注射Ad-GFAP—hNIS、不注射131I，注射Ad—GFAP—hNIS、注射131I，注射Ad—CMV—EGFP、不注射131I，注射Ad—CMV—EGFP、注射131I，每组5只），各组进行相应处理后观察移植瘤生长情况并比较裸鼠生存周期；U87荷瘤裸鼠肿瘤内注射Ad—GFAP—bNIS和Ad．CMV—EGFP后腹腔内注射1mCi的^99mTcO4溶液，应用SPECT显像。结果与PGL3-Basic、PGL3．Control比较，PGL3-GFAP质粒转染后U251、U87细胞相对反应活性降低，差异有统计学意2Z（P〈0．05）；Western blotting显示U87和U251细胞表达GFAP、hNIS蛋白；125I摄碘能力试验显示感染重组腺病毒Ad-GFAP—hNIS后，U87和U251细胞摄碘能力分别提高69．5倍和70．8倍，细胞内131I的有效半衰期缩短；在U87和U251细胞中，Ad-GFAP-hNIS组细胞克隆形成率分别为（9.31±0．50）％和（9．33±1．15）％，均高于Ad．CMV．EGFP组和Ad—CMV—hNIS组，差异有统计学意义（P〈0．05）；注射Ad—GFAP．hNIS且注射131I组裸鼠生存周期（44．00±0．58）d最长；SPECT实验显示Ad-GFAP—hNIS组裸鼠肿瘤组织可以摄取放射性核素，而Ad—CMV—EGFP组裸鼠不能摄取。结论感染Ad-GF

英文摘要：

Objective To fred the possibility that the glial fibrillary acidic protein （GFAP） promoters modulate the human sodium iodide symporter （hNIS） expression in glioma and lead transecting hNIS gene into glioma cells for radioactive iodide treatment. Methods PGL3-Basic, PGL3-Control and PGL3-GFAP plasmids were transfected into U251, U87 and MRC-5 cells, respectively, with the help of liposome Lipofectamine 2000; 24 h after that, the reactivity of these cells was detected and the efficiency of GFAP promoter was tested under chemiluminescence apparatus. Recombinant adenovirus vector Ad-GFAP-hNIS in which GFAPpromoter could modulate the hNIS gene expression was constructed, and then, the vector was transfected into the U251, U87 and MRC-5 cells; Western blotting was employed to detect the protein expressions of GFAP and hN1S. Ad-CMV-EGFP group （blank control） and Ad-CMV-hNIS group （negative control） and Ad-GFAP-hNIS group were employed; the 131I uptake and effiux abilities and the cell amount after gentian violet staining in the three groups were measured by counter; the clonogenecity rate of them was calculated. BALB/c female nude mice （n=20） was divided into four groups： group of injecting Ad-GFAP-hNIS without ~31I, group of injecting Ad-GFAP-hNIS with 131I, group of injecting Ad-CMV-EGFP without 13q and group of injecting Ad-CMV-EGFP with 131I （n=5）; U87 cells were transfected into the nude mice, and then, the tumor growth was observed and the life cycle of the mice was noted. Nude mice bearing the U87 tumors were injected Ad-GFAP-hNIS and Ad-CMV-EGFP, followed by 1mCi ^99TcO4 via intraperitoneal injection; single photon emission computed tomography （SPECT） was performed. Results As compared with that of cells being transfected with PGL3-blank plasmid, the relative reactivity of U251 and U87 cells being transfected with PGL3-GFAP plasmid was decreased with significant difference （P〈0.05）. Western blotting revealed GFAP and hNIS proteins in U87 and U251 cells. 125I uptake of U87 and