胰腺癌多药耐药细胞株BxPC～3／ADM的建立及耐药机制的初步探讨  

Construction of multidrug resistant cell line BxPC-3/ADM of pancreatic cancer and initial investigation of its resistance mechanism

作　　者：蒋永剑[1] 虞先浚[1] 徐近[1] 李骥[1] 傅德良[1] 倪泉兴[1] 

机构地区：[1]复旦大学附属华山医院外科 ,上海200040

出　　处：《中华肝胆外科杂志》2009年第11期835-838,共4页Chinese Journal of Hepatobiliary Surgery

摘　　要：目的构建胰腺癌多药耐药细胞株BxPC-3／ADM，通过动态监测诱导耐药过程，探讨其耐药的可能机制。方法逐步增加阿霉素浓度对BxPC-3细胞进行诱导，在不同的阿霉素诱导浓度，MTT法检测细胞对多种化疗药物的耐药指数，RTPCR和Western印迹法检测细胞MDR1和MRPmRNA和蛋白的表达。结果成功构建多药耐药细胞株BxPC-3／ADM；在0．05μg／ml至8μg／ml阿霉素诱导浓度，细胞对5-Fu、MMC和Gem的耐药指数无明显增加；在16μg／ml浓度3种化疗药物的耐药指数有较明显上升，同时伴有MDR1mRNA表达的明显增加；至32μg／ml 5-Fu和Gem的耐药指数未再有继续上升，而MMC的耐药指数则继续上升，同时MDR1mRNA表达未再增加，而MRPmRNA表达则明显增加。Western印迹实验显示MDR1和MRP蛋白表达变化与mRNA表达变化趋势一致。结论MDR1基因在诱导早期的耐药中起主导作用，在后期则和MRP基因协同发挥作用。Objective To construct multidrug resistance cell line BxPC-3/ADM of pancreatic cancer and investigate its resistance mechanism through dynamically monitoring the inducing course. Methods BxPC-3 cell was induced by increasing the concentration of ADM gradually. At the different inducing concentrations of ADM, the resistance indexes to multiple chemotherapeutics were detected by MTT and the mRNA and protein expression of MDR1 and MRP were determined by RT-PCR and Western blot, respectively. Results Multidrug resistant cell line BxPC-3/ADM was constructed suc cessfully. When the inducing concentration of ADM was from 0.05 μg/ml to 8 /μg/ml, the resistance indexes to 5-Fu, MMC and Gem did not significantly increase. When the inducing concentration of ADM was 16μg/ml, the resistance indexes to three chemotherapeuties increased significantly and the mRNA expression of MDR1 obviously rose simultaneously. When the inducing concentration of ADM was 32 μg/ml, the resistance indexes to 5-Fu and Gem did not increase continually, whereas the resistance index to MMC increased continually. Meanwhile, the mRNA expression of MDR1 did not keep on rising, whereas the mRNA expression of MRP obviously increased. Western blot showed that the change of MDR1 and MRP protein expression was consistent with the change of mRNA expression. Conclusion During the inducing course, MDR1 has its main contribution at the early stage, whereas MDR1 and MRP have co-contributing action at the late stage.

关 键 词：胰腺肿瘤 多药耐药 MDR1 MRP 

分 类 号：R735.9[医药卫生—肿瘤]