突变型ⅠκBα转染对肝门部胆管癌多药耐药基因的调控作用

Effect of mⅠκBα transfection on multidrug resistance gene in hilar cholangiocarcinoma cell lines

目的 探讨阻断核转录因子 κB(NF κB)活性对肝门部胆管癌细胞多药耐药基因(MDR 1)表达的影响。方法 用突变核转录因子 κB抑制蛋白αⅠκBα质粒 (mⅠκBα)转染肝门部胆管癌细胞株 (QBC93 9)及稳定表达丙型肝炎病毒核心蛋白 (HCVC)的QBC93 9(QBC93 9HCVC+) ,凝胶迁移率电泳 (EMSA)检测mⅠκBα质粒转染的QBC93 9和QBC93 9HCVC +细胞中NF κBDNA结合活性 ;逆转录 多聚酶链反应 (RT PCR )检测转染mⅠκBα质粒对肝门部胆管癌细胞中MDR 1基因及其表达产物 (P GP )表达的影响。结果 转染mⅠκBα质粒组的QBC93 9和QBC93 9HCVC +细胞放射性浓聚影明显淡于非转染组 ,密度计成对扫描显示 ,转染组和非转染组相对信号密度有明显差异 ;转染突变型ⅠκBα质粒的QBC93 9和QBC93 9HCVC +细胞中MDR 1mRNA和蛋白的表达水平明显低于非转染组。结论 转染mⅠκBα能明显逆转肝门部胆管癌细胞多药耐药性 ,阻断NF κB活性有望成为肝门部胆管癌基因治疗新策略的“靶位”。

Objective To explore the effect on the expression of multidrug resistance gene and protein in hilar cholangiocarcinoma cell linves by inhibiting NF-κB.Methods QBC939HCVC+ cells and QBC939 cells were transfected by mutated ⅠκB plasmid.The activated NF-κB was detected by EMSA,and RT-PCR and immunohistochemical assay were used to detect the expression of MDR-1 mRNA and protein (P-GP) expression.Results The NF-κB DNA binding activity of QBC939HCVC+ and QBC939 cells in transfecting mutated ⅠκB plasmid was lower than in the untransfecting mutated ⅠκB plasmid(P<0.01).The expession of MDR-1mRNA and P-GP was significantly decreased by transfecting mutated ⅠκB plasmid (P<0.01).Conclusion Transfecting mutated ⅠκB plasmid can deteriorate multidrug resistance of hilar cholangiocarcinoma cells.Interruption of NF-κB maybe come a new target in the gene therapy aimed to hilar cholangiocarcinogenesic carcinoma.