JNK通路介导人肝癌耐药细胞株Bel-7402/FU多药耐药的调控机制  

The regulatory mechanism of JNK signal transduction pathway-mediated multidrug-resistance in human hepatic cancer cell line Bel-7402/FU

作　　者：黄灿[1,2] 许杜娟[3] 居靖[1] 夏泉[3] 王猛[2] 

机构地区：[1]安徽医科大学附属安庆医院药剂科,安徽安庆246000 [2]安徽医科大学药学院,安徽合肥230032 [3]安徽医科大学第一附属医院药剂科,安徽合肥230022

出　　处：《肿瘤》2014年第1期19-25,共7页Tumor

基　　金：安徽省自然科学基金资助项目(编号:11040606M222)

摘　　要：目的:探讨C-Jun氨基末端激酶(C-Jun N-terminal kinase,JNK)信号转导通路对人肝癌5-氟尿嘧啶(5-fluorouracil,5-FU)耐药细胞系Bel-7402/FU产生多药耐药(multidrug resistance,MDR)的调控作用,为肝癌细胞MDR机制的研究提供新靶点。方法:采用蛋白质印迹法检测人肝癌亲本细胞Bel-7402和耐药细胞Bel-7402/FU中JNK及磷酸化-JNK(phospho-JNK,p-JNK)的表达;在Bel-7402/FU细胞中,使用特异性阻断剂SP600125抑制TNK通路后,采用RT-PCR和免疫细胞化学法分别检测MDR1 mRNA和P-糖蛋白(P-glycoprotein,P-gp)的表达;FCM法检测罗丹明123(rhodanmine 123,Rh123)在细胞内的积聚和外排情况;MTT法检测Bel-7402/FU细胞对5-FU的敏感性。结果:耐药细胞Bel-7402/FU组与亲本细胞Bel-7402组相比,JNK蛋白的表达无明显变化,而p-JNK蛋白的表达量明显增加;抑制JNK通路后,Bel-7402/FU细胞中MDR1 mRNA表达水平和P-gP蛋白的表达水平均明显下调,Rh123的蓄积明显增加,外排明显减少,且对5-FU的敏感性明显增加。结论:JNK信号转导通路参与人肝癌耐药细胞株Bel-7402/FU中多药耐药基因MDR1和多药耐约蛋白P-gp的表达,对其耐药起调控作用。Objective： To investigate the regulatory effect of C-Jun N-terminal kinase （JNK） signal transduction pathway on multi-resistance in the human hepatic cancer Bel-7402/5-fluorouracil （FU） cells, and provide a possible novel target for study on the mechanism of multidrug-resistance in human hepatic cancer cells. Methods： The protein expression levels of JNK and phospho-JNK （p-JNK） in parental human hepatic cancer Bel-7402 cells and the drug-resistant Bel-7402/FU cells were detected by Westem blotting. After the inhibition of JNK pathway induced by specific inhibitior SP600125, the expressions of multi-drug resistance 1 （MDR1） mRNA and P-glycoprotein （P-gp） in Bel-7402/FU cells were detected by RT-PCR and immunocytochemistry, respectively. The accumulation and efflux of rhodanmine 123 （Rh123） in the cells were examined by flow cytometry, and the sensitivity to 5-FU of Bel-7402/FU cells was detected by MTT method. Results： The expression of JNK protein was not significantly different between the Bel-7402/FU cells and the parental Bel-7402 cells, but the expression of p-JNK protein in the Bel-7402/FU cells was significantly increased. After inhibition of JNK pathway, the expressions of MDR1 mRNA and P-gp protein were obviously decreased, with a markedly increased accumulation and decreased efflux of Rh123, leading to enhhanced sensitivity to 5-FU of Bel-7402/FU cells. Conclusion： JNK signal transduction pathway is involved in the expressions of MDR1 gene and P-gp in drug-resistant human hepatic cancer Bel-7402/FU cells, and it can regulate the sensitivity to 5-FU of Bel-7402/FU cells.

关 键 词：肝肿瘤 多药耐药相关蛋白质类 JNK信号通路 BEL-7402 FU细胞 

分 类 号：R735.7[医药卫生—肿瘤]