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张宏飞

作品数:3 被引量:2H指数:1
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脂质体介导人NIS基因转染肺癌A549细胞及其蛋白表达被引量:1
2012年
目的研究人钠/碘同向转运体(NIS)基因转染肺癌细胞及其蛋白表达。方法鉴定质粒pcDAN3-hNIS中的插入基因NIS基因。培养的肺癌A549分为两组:实验组(转染pcDAN3-hNIS),对照组(转染pcDAN3)。脂质体介导NIS基因转染肺癌细胞,采用Western Blot免疫印迹法和免疫组化法检测肺癌细胞中NIS蛋白的表达。结果实验组的肺癌细胞有NIS蛋白的表达,而对照组无表达,两组比较差异有显著性(P=0.000)。结论转染人NIS基因的肺癌细胞可表达NIS蛋白,为探索放射性碘治疗肺癌的研究提供理论依据。
张宏飞严煜张裕东龚德军
关键词:肺癌细胞基因转染脂质体放射性碘治疗
Transfection of the Human Sodium/Iodide Symporter(NIS) Gene with Liposomes and the Expression of the NIS Protein in Human Lung A549 Cancer Cells被引量:1
2008年
OBJECTIVE To examine the possibility of human sodium iodide symporter (hNIS) protein expression in lung cancer cells. METHODS Human lung A549 cancer cells were thawed and cultured in vitro. The cells were divided into an experimental group transfected with a recombinant pcDNA3-hNIS plasmid and a control group transfected only with a pcDNA3 plasmid. The recombinant plasmid vector encoding the hNIS gene (pcDNA3-hNIS) was amplified, purified and identified. The hNIS gene was followed by DNA sequencing. A Western blot and an immunohistochemical assay were applied to detect the hNIS protein expression in the transfected human lung A549 cancer cells. RESULTS Restriction enzyme digestion and DNA sequencing results showed the size and direction of the inserted gene in the recombinant pcD- NA3-hNIS plasmid was correct. The Western blot method and immunohistochemical analysis showed a positive NIS protein expression in the experimental group. The NIS protein was detected mainly in the cell membranes showing a positive rate up to 70.6% with no expression of the NIS protein in the control group. There was a significant difference between two groups (P=0.000). CONCLUSION The hNIS gene was transfected effectively into human lung A549 cancer cells mediated by Lipofectamine 2000, and was expressed with its protein in vitro.
严煜张宏飞张裕东王晓谭
关键词:LIPOSOME
人重组pcDNA3人钠/碘同向转运体核表达质粒的构建
2006年
目的:获取、构建并鉴定携带人钠/碘同向转运体(hNIS)基因的pcDNA3.0质粒载体。方法:利用逆转录和聚合酶链反应获取Graves甲亢患者甲状腺NIS基因可编码区片段。与pUCm-T质粒载体连接、经转化、蓝白斑筛选、酶切鉴定,序列测定鉴定,获得pUCm-T-重组质粒。pUCm-T-重组质粒亚克隆得到pcDNA3-hNIS重组质粒。结果:所构建的pcDNA3-hNIS重组质粒中的hNIS基因序列与文献报道序列一致。结论:成功构建pcDNA3-hNIS重组质粒。
张宏飞严煜张裕东
关键词:基因转移逆转录聚合酶链反应
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