JNK信号通路介导MDR1/P-gp调控人结肠癌多药耐药  

JNK signal transduction pathway regulates MDR1/P-glycoprotein-mediated multidrug resistance in colon carcinoma cells

作　　者：隋华[1,2] 周利红[1,2] 殷佩浩[1,2] 王炎[1,2] 范忠泽[1,2] 周树峰 李琦[1,2] 

机构地区：[1]上海中医药大学附属普陀医院 [2]上海中医药大学中西医结合肿瘤介入研究所,上海市200062 [3]南佛罗里达大学药学院,美国佛罗里达州坦帕市33612

出　　处：《世界华人消化杂志》2011年第9期892-898,共7页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.308734;上海市科委基金资助项目;No.10ZR1427400;No.10140902600;上海市教委科研创新基金资助项目;No.09YZ132;上海市重点学科基金资助项目;No.S30302~~

摘　　要：目的:探讨在人结肠癌多药耐药细胞株HCT8/V中JNK信号传导通路与多药耐药1(MDR1)基因表达的关系.方法:MTT法检测人结肠癌多药耐药细胞株HCT8/V及其敏感株HCT8细胞对多种抗肿瘤药物的敏感性;抑制JNK信号通路的激活后,采用双荧光素酶活性报告基因检测MDR1启动子转录活性的表达;实时荧光定量PCR检测MDR1mRNA的表达;Westernblot分析对P-糖蛋白(P-gp)表达的影响.结果:HCT8/V细胞对长春新碱(VCR)的耐药倍数为10.49倍.加入JNK特异性抑制剂sp600125(20μmol/L)抑制后,HCT8/V细胞对VCR的半数抑制浓度(IC50)由191.08mg/L±18.18mg/L降低到50.34mg/L±15.71mg/L,逆转指数为3.80(P<0.01).HCT8/V细胞加入JNK抑制剂后,MDR1启动子活性从0.03647±0.00191降低到0.01362±0.00196,下降了2.52倍(P<0.01).实时荧光定量PCR、Westernblot检测结果提示,抑制JNK通路的激活后,HCT8/V细胞的MDR1mRNA从0.34275±0.0339下降到0.13625±0.0196(P<0.01),P-gp表达从0.88132±0.1026下降到0.56174±0.1014(P<0.01).结论:JNK信号通路通过介导MDR1/P-gp调控人肠癌多要耐药,抑制JNK信号通路可降低MDR1/P-gp的水平,逆转人结肠癌HCT8/V细胞的多药耐药性.AIM：To investigate the relationship between the JNK（c-Jun NH2-terminal kinase） signal transduction pathway and multidrug resistance 1（MDR1）/P-glycoprotein（P-gp）-mediated multidrug resistance of human colon carcinoma HCT8/V cells.METHODS：The sensitivity of multidrug-resistant HCT8/V cells and drug-sensitive HCT8 cells to a variety of antitumor drugs was evaluated by MTT assay.After inhibition of the activation of the JNK signaling pathway,MDR1 promoter transcriptional activity and expression of MDR1 mRNA and P-gp were evaluated by luciferase activity assay,real-time fluorescence quantitative PCR（RFQ-PCR） and Western blot,respectively.RESULTS：HCT8/V cells exhibited 10.49-fold higher resistance to vincristine than their parental counterparts.The IC50 value of SP600125（20 μmol/L） decreased from 191.08 mg/L ± 18.18 mg/L to 50.34 mg/L ± 15.71 mg/L and resistance index（RI） reached 3.80（P 0.01）.After inhibition of the activation of the JNK signaling pathway with a JNK specific inhibitor,MDR1 promoter transcriptional activity showed a 2.55-fold decrease（0.03647 ± 0.00191 vs 0.01362 ± 0.00196,P 0.01） and the expression of MDR1/P-gp was significantly downregulated（0.34275 ± 0.0339 vs 0.13625 ± 0.0196,0.88132 ± 0.1026 vs 0.56174 ± 0.1014,both P 0.01） in HCT8/V cells.CONCLUSION：The JNK signal transduction pathway regulates MDR1/P-glycoprotein-mediated multidrug resistance of HCT8/V cells possibly by up-regulating MDR1/P-gp expression.

关 键 词：结肠癌 多药耐药1 P-糖蛋白 C-JUN氨基末端激酶 信号转导通路 

分 类 号：R735.35[医药卫生—肿瘤]