Chao C C
Department of Biochemistry, Chang Gung Medical College, Taoyuan, Taiwan, Republic of China.
FEBS Lett. 1995 Oct 16;373(3):285-90. doi: 10.1016/0014-5793(95)01063-k.
A multidrug-resistant (MDR) subline of the immunoblastic B lymphoma cell line was established by sequentially selecting in increasing concentrations of adriamycin. The adriamycin-resistant cell line (HOB1/ADR) demonstrated resistance to a wide spectrum of chemotherapeutic agents including MDR drugs (Vinca alkaloids and anthracycline), antimicrotubule drug (colchicine), and DNA-damaging agents (cisplatin and mitomycin C). The expression of human mdr1 gene, as analyzed by RT-PCR and Western blotting, revealed a 13-15-fold increase in resistant cells. Unexpectedly, HOB1/ADR cells demonstrated a lack of reduced accumulation and of enhanced efflux of adriamycin. More than 60% adriamycin was effluxed at the same rate in both cell lines within 10 min. In contrast, the initial rate of vincristine accumulation was reduced by 3 fold in this resistant cell line. The maximal level of vincristine accumulation was 50% lower in the resistant cells than the parental cells. The maximal efflux rate was enhanced by 5 fold in the resistant cells. Inhibition of vincristine resistance by verapamil associated with restoration of drug accumulation, suggesting that acquired resistance in these cells is due to P-glycoprotein. These studies demonstrated that immunoblastic B lymphoma cells selected for adriamycin resistance preferentially developed P-glycoprotein-mediated vincristine efflux which plays an important role in vincristine resistance. In contrast, the resistant cells did not elevate adriamycin efflux, suggesting an additional mechanism responsible for adriamycin resistance.
通过在逐渐增加浓度的阿霉素中依次筛选,建立了免疫母细胞性B淋巴瘤细胞系的多药耐药(MDR)亚系。阿霉素耐药细胞系(HOB1/ADR)对多种化疗药物具有抗性,包括MDR药物(长春花生物碱和蒽环类药物)、抗微管药物(秋水仙碱)和DNA损伤剂(顺铂和丝裂霉素C)。通过RT-PCR和蛋白质免疫印迹分析人mdr1基因的表达,结果显示耐药细胞中该基因表达增加了13 - 15倍。出乎意料的是,HOB1/ADR细胞对阿霉素的摄取减少和外排增强并不明显。在10分钟内,两种细胞系中超过60%的阿霉素以相同速率外排。相比之下,该耐药细胞系中长春新碱的初始摄取速率降低了3倍。耐药细胞中长春新碱的最大摄取水平比亲代细胞低50%。耐药细胞中长春新碱的最大外排速率提高了5倍。维拉帕米抑制长春新碱耐药并伴随着药物摄取的恢复,这表明这些细胞获得性耐药是由于P-糖蛋白所致。这些研究表明,经阿霉素耐药筛选的免疫母细胞性B淋巴瘤细胞优先产生P-糖蛋白介导的长春新碱外排,这在长春新碱耐药中起重要作用。相比之下,耐药细胞并未提高阿霉素外排,提示存在另一种导致阿霉素耐药的机制。