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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
Objective To investigate the expression of human sodium / iodide symporter (hNIS) gene in human cholangiocarcinoma cell line QBC939 and its effect on the uptake of 125I. Methods The cDNA sequence of hNIS gene was cloned into pMD18-T vector. The selected subclone hNIS was cloned into eukaryotic expression vector PDC316 and introduced into cholangiocarcinoma cell line QBC939 by lipofectamine to establish a new cell line (QBC939-A). At the same time, empty plasmid (QBC939-B) transfection and blank (QBC939-C) control group were established. Semi-quantitative RT-PCR was used to detect the expression of hNIS-mRNA and the uptake of 125I on 1,2,3,6 d in each group. Results A novel cell line QBC939-A that can stably express hNIS gene was established. The expression of hNIS-mRNA showed that the expression of QBC939-A peaked at 3 days, which was significantly different from that of QBC939-B and QBC939-C (P <0.05). The uptake of iodine by QBC939-A cells peaked at 3 days after transfection, which was 14 times higher than that of QBC939-C and 16 times higher than that of QBC939-B. CONCLUSIONS: rhNIS transduction into cholangiocarcinoma cells is sufficient to mediate the uptake of 125I in the short term, which is the basis for the study of 125I-targeted cholangiocarcinoma