阿霉素在人白血病敏感细胞株HL-60和多药耐药细胞株HL-60/ADR的分布和积聚变化

目的了解化疗药物阿霉素(ADR)在人白血病敏感细胞株HL-60和多药耐药细胞株HL-60/ADR细胞内的分布和积聚变化及其与耐药的关系。方法应用共聚焦激光扫描显微镜和流式细胞术研究ADR在细胞内的分布和积聚,以及维拉帕米、BSO、布雷菲尔得菌素、氯喹对细胞内ADR异常分布的影响。以3种特异染色线粒体、高尔基体和溶酶体的荧光物质Rhodamine123、NBD-ceramide和中性红作为探针,鉴定分隔储留ADR的细胞器。结果与ADR在HL-60细胞核、胞质均匀分布不同,在HL-60/ADR,ADR呈点棘状分布于细胞质,核内ADR荧光很少。这种分布方式与NBD-ceramide荧光在该细胞的分布极其相似。BSO和布雷菲尔得菌素可逆转ADR在HL-60/ADR的异常分布和积聚,而维拉帕米和氯喹无此作用。结论ADR在耐药细胞的异常分布和积聚与肿瘤细胞耐药的形成有关。

Changes of the distribution and accumulation of adriamycin in the sensitive and multidrug-resistant HL-60 cells

Aim To investigate subcellular distribution and accumulation of ADR in the sensitive and multidrug-resistant HL-60 cells and its relation to muhidrug resistance. Methods The subcellular distribution and accumulation of ADR were studied by confocal scanning laser microscope and flow cytometry. The effects of verapamil, BSO, brefeldin A and chloroquine on ADR distribution and accumulation in HL-60/ADR cells were also examined. Rhodamine123, NBD-ceramide and neutral red were used as fluorescent the mitochondria, Golgi apparatus and lysosomes respectively were used to identify the subcellular compartments where ADR was sequestered. Results In drug-sensitive cell line HL-60, ADR fluorescence distributed evenly in the nucleus and cytoplasm, while in muhidrug-resistant cell line HL-60/ADR, ADR fluorescence distributed in a punctated pattern in the cytoplasm and was reduced in the nucleus. The mode of ADR distribution in HL-60/ADR cells is highly similar to that of NBD-ceramide. BSO and brefeldin A,instead of verapamil and chloroquine could reverse the abnormal distribution and accumulation of ADR in HL-60/ ADR cells. Conclusions The change of ADR distribution and reduction of ADR accumulation in muhidrug-resistant cell line was involved in the mechanism of muhidrug resistance.