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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。
Objective To investigate the distribution and accumulation of chemotherapeutic drug adriamycin (ADR) in human leukemia cell line HL-60 and multidrug-resistant cell line HL-60 / ADR and its relationship with drug resistance. Methods Confocal laser scanning microscopy and flow cytometry were used to investigate the distribution and accumulation of ADR in cells and the effects of verapamil, BSO, brefeldin and chloroquine on the abnormal distribution of intracellular ADR. Rhodamine123, NBD-ceramide and neutral red, fluorescent substances of mitochondria, Golgi and lysosomes, were used as probes to identify organelles that separate and retain ADRs. Results Compared with ADR, the nuclear and cytoplasmic distribution of HL-60 cells was different from that of ADR. In HL-60 / ADR, the ADR showed some spiny distribution in the cytoplasm with little ADR fluorescence in the nucleus. This distribution is very similar to the distribution of NBD-ceramide fluorescence in this cell. BSO and brefeldin could reverse the abnormal distribution and accumulation of ADR in HL-60 / ADR, whereas verapamil and chloroquine did not. Conclusion The abnormal distribution and accumulation of ADR in drug-resistant cells is related to the formation of drug-resistant tumor cells.