rhNIS基因转导胆管癌细胞介导125I摄取的研究

Study on the transfer of the rhNIS gene into human bile duct carcinoma cells to induce125I uptake

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。

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.