Neyfakh A A
Interfaculty Laboratory of Molecular Biology and Bioorganic Chemistry, Moscow State University, USSR.
Exp Cell Res. 1988 Jan;174(1):168-76. doi: 10.1016/0014-4827(88)90152-8.
Fluorescence microscopy has shown that 18 different fluorescent dyes, staining various intracellular structures in transformed hamster fibroblasts (DM-15), did not stain or stained weakly multidrug-resistant cells selected from DM-15 by colchicine. Reduced staining by fluorescent dyes was characteristic also of five other tested multidrug-resistant cell lines of hamster and mouse origin, selected by actinomycin D, colcemid, rubomycin, and ruboxyl. The intensity of staining of two revertant cell lines was similar to that of parental sensitive cells. All tested inhibitors of multidrug resistance, including weak detergent, metabolic inhibitors, calcium channel blockers, calmodulin inhibitors, and reserpine, restored normal staining of multidrug-resistant cells. The dyes accumulated in resistant cells in presence of these inhibitors left the cells several minutes after the removal of the inhibitor from the incubation medium. Sensitive cells retained the dyes for several hours. The efflux of the dyes from resistant cells is an active process since it occurred even in the presence of the dyes in the incubation medium. The efflux could be blocked by all tested inhibitors of multidrug resistance and it is possibly a basic mechanism of the reduced staining of resistant cells. These data support the idea that multidrug resistance is based on active nonspecific efflux of the drugs and indicate that the simple procedure of cell staining can be used for the detection of resistant cells and further study of the phenomenon of multidrug resistance.
荧光显微镜检查显示，18种不同的荧光染料可对转化的仓鼠成纤维细胞（DM - 15）中的各种细胞内结构进行染色，但对通过秋水仙碱从DM - 15中筛选出的多药耐药细胞不染色或染色较弱。荧光染料染色减少也是其他5种经测试的仓鼠和小鼠来源的多药耐药细胞系的特征，这些细胞系是通过放线菌素D、秋水仙酰胺、柔红霉素和鲁保菌素筛选出来的。两个回复细胞系的染色强度与亲代敏感细胞相似。所有测试的多药耐药抑制剂，包括弱去污剂、代谢抑制剂、钙通道阻滞剂、钙调蛋白抑制剂和利血平，均可恢复多药耐药细胞的正常染色。在这些抑制剂存在的情况下，耐药细胞中积累的染料在从孵育培养基中去除抑制剂几分钟后就会离开细胞。敏感细胞可将染料保留数小时。染料从耐药细胞中的流出是一个活跃过程，因为即使在孵育培养基中存在染料时也会发生。流出可被所有测试的多药耐药抑制剂阻断，这可能是耐药细胞染色减少的基本机制。这些数据支持了多药耐药基于药物的主动非特异性流出这一观点，并表明细胞染色的简单程序可用于检测耐药细胞以及进一步研究多药耐药现象。
