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rhNIS基因转导胆管癌细胞介导125I摄取的研究
引用本文:张先林,丁志强,陈勇军,俞亚红,杨继鑫,陈忠. rhNIS基因转导胆管癌细胞介导125I摄取的研究[J]. 中国普通外科杂志, 2009, 18(5): 9-472
作者姓名:张先林  丁志强  陈勇军  俞亚红  杨继鑫  陈忠
作者单位:(华中科技大学同济医学院附属同济医院 胆胰外科, 湖北 武汉 430030)
摘    要:
目的:探讨人钠/碘同向转运体(hNIS)基因cDNA转导至胆管癌细胞系QBC939细胞后的表达水平及其对介导125I摄取的影响。方法:将扩增出的hNIS基因cDNA序列克隆至pMD18-T载体;筛选后的亚克隆hNIS编码至真核表达型载体PDC316中,经脂质体途径导入胆管癌细胞(QBC939),建立新细胞系(QBC939-A) 。同时设立空质粒(QBC939-B)转染和空白(QBC939-C)对照组。在体外培养条件下采用半定量RT-PCR检测各组2,3,6 d细胞内hNIS-mRNA表达水平和1,2,3,6 d的125I摄取情况。结果:建立了能稳定表达hNIS基因的新型细胞系QBC939-A。hNIS-mRNA表达水平检测显示,QBC939-A于3 d表达量达高峰,与QBC939-B和QBC939-C比较,有显著性差异(P<0.05)。且QBC939-A细胞的摄碘能力在转染后3 d达高峰,较QBC939-C高14倍,较QBC939-B高16倍。结论:rhNIS基因转导至胆管癌细胞足以在短期内介导125I的摄取,为介导125I靶向治疗胆管癌的研究奠定了基础。

关 键 词:胆管肿瘤; 钠/碘转运体; 反转录-多聚酶链反应; 重组质粒; 胆管癌细胞; 碘放射性同位素
收稿时间:1900-01-01
修稿时间:1900-01-01

Study on the transfer of the rhNIS gene into human bile duct carcinoma cells to induce125I uptake
ZHANG Xianlin,DING Zhiqiang,CHEN Yongjun,YU Yahong,YANG Jixin,CHEN Zhong. Study on the transfer of the rhNIS gene into human bile duct carcinoma cells to induce125I uptake[J]. Chinese Journal of General Surgery, 2009, 18(5): 9-472
Authors:ZHANG Xianlin  DING Zhiqiang  CHEN Yongjun  YU Yahong  YANG Jixin  CHEN Zhong
Affiliation:(Department of Biliary and pancreatic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030,China)
Abstract:
Objective:To study the expression level and effect of hNIS gene to induced125I uptake after transfecting hNIS gene into cholangiocarcinoma cell line QBC939. Methods :The hNIS cDNA sequences by amplification were ligated with plasmid pMD18-T, selected, subcloned and ligated with eucaryotic expressing plasmid PDC316, using lipofectamine to transfect eucaryotic expressing plasmid vector hNIS-DNA-PDC316(experimental group)and plasmid PDC316(control group)into cholangiocarcinoma cell line QBC939, establishing new cell lines(QBC939-A,QBC939-B) and QBC939-C (control).The expression level of hNIS gene was detected by semi-quantitative RT-PCR in 2-, 3-, and 6-day, and the effect of induced125I uptaking was studied on the 1-,2-, 3-, and 6 day in cell lines of vitro culture.Results:QBC939-A stably expressed hNIS-DNA-PDC316.The expression level on QBC939-A compared with QBC939-B and QBC939-C had significant difference (P<0.05), the peak of induced125I uptaking was detected on the 3rd day in QBC939-A, and cell line QBC939-A accumulated 14 and 16 times more radio-iodide in vitro than QBC939-C and QBC939-B respectively. Conclusions:Tramsfer of hNIS gene into cholangiocarcinoma cells can induce125I uptake in the short-term, This study can be as a foundation for study of targeting action to bile duct carcinoma using radionuclide125I.
Keywords:Bile Duct Neoplasms   Sodium/Iodide Symporter   RT-PCR   Recombinant Plasmid, Cholangiocarcinoma Cell   Iodine Radioisotope
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