Vesicular anthracycline accumulation in Doxorubicin-selected U-937 cells: Participation of lysosomes
Vesicular anthracycline accumulation in Doxorubicin-selected U-937 cells: Participation of lysosomes
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DOI：
10.1182/blood.v89.10.3745.3745_3745_3754
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发表时间：
1997-05-15
期刊：
影响因子：
20.3
通讯作者：
Slapak, CA
中科院分区：
文献类型：
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作者：
Hurwitz, SJ;Terashima, M;Slapak, CA
U-A10细胞系是人类U-937髓系白血病细胞的阿霉素选择变体，表现出蒽环类药物在扩大的囊室中的重新分布。通过pH敏感染料LysoSensor黄/蓝DND-160的活体染色证实了该隔室的酸性。通过免疫荧光分析进行囊泡室的鉴定。LAMP-1和LAMP-2抗原染色显示，囊泡是增大的溶酶体，位于U-A10细胞细胞核附近。多药耐药相关蛋白和p -糖蛋白主要在耐药细胞的质膜上表达。柔红霉素在细胞间室的积累是用放射性标记的药物定量。将细胞暴露在(3)[H]-柔红霉素中，然后分离完整的细胞核，U-A10细胞的细胞核比U-937细胞的细胞核少2 - 3倍的蒽环类药物积累，而先分离细胞核再暴露在(3)[H]-柔红霉素中，细胞核的净药物积累差异不大。将U-A10和U-937细胞制备的细胞质暴露于(3)[H]柔红霉素中，测定细胞质药物积累。
The U-A10 cell line, a doxorubicin-selected variant of human U-937 myeloid leukemia cells, exhibits a redistribution of anthracyclines into a expanded vesicular compartment. The acidic nature of this compartment was confirmed by vital staining with a pH sensitive dye, LysoSensor yellow/blue DND-160. Identification of the vesicular compartment was performed by immunofluorescence analysis. Staining for the LAMP-1 and LAMP-2 antigens showed that the vesicles are enlarged lysosomes that are eccentrically placed near the nucleus of U-A10 cells. By contrast, the expression of the multidrug resistance-associated protein and the P-glycoprotein wits observed predominately on the plasma membrane of the drug-resistant cells. The accumulation of daunorubicin into cellular compartments was quantified using radiolabeled drug. Exposing cells to (3)[H]-daunorubicin and then isolating intact nuclei showed that nuclei from U-A10 cells accumulated twofold to threefold less anthracycline than nuclei from U-937 cells, However, when nuclei were isolated first and then exposed to (3)[H]-daunorubicin, little difference in net nuclear drug accumulation was detected. Cytoplasts prepared from U-A10 and U-937 cells were exposed to (3)[H]daunorubicin to measure cytoplasmic drug accumulation.