Altered intracellular distribution of daunorubicin in immature acute myeloid leukemia cells.
We have used laser-assisted confocal microscopy to evaluate the intracellular distribution of daunorubicin (DNR) in acute myeloid leukemia (AML) cell lines and fresh AML cells according to their differentiation phenotype. In KG1a, KG1, TF-1 and HEL cells, which express the early differentiation marker CD34, DNR was distributed in perinuclear vesicles which could be associated with the Golgi apparatus, as suggested by the distribution of fluorescent probes specific for intracellular organelles. In contrast, U937 and HL-60 cells, which display a more mature phenotype, exhibited nuclear and diffuse cytoplasmic DNR fluorescence. DNR sequestration was not correlated with P-glycoprotein (P-gp) or multidrug resistance protein expression. Furthermore, PSC833, a potent P-gp blocker, had little effect on drug sequestration in CD34+ AML cells. We also tested the effect of metabolic inhibitors, cytoskeleton inhibitors and carboxy-ionophores on DNR distribution in both CD34- and CD34+ AML cells. However, only non-specific metabolic inhibitors restored nucleic/cytoplasmic distribution in CD34+ cells. In these cells, the intracellular distribution of doxorubicin and idarubicin was very similar to that of DNR, while the distribution of methoxymorpholinyl-doxorubicin was nuclear and diffusely cytoplasmic. In fresh AML cells, DNR was also concentrated in the perinuclear region in CD34+ but not in CD34- cells. However, DNR sequestration was not observed in normal CD34+ cells. Finally, our results show that DNR is sequestered in organelles in CD34+ AML cells via an active mechanism which appears to be different from P-gp-mediated transport. Abnormal DNR distribution may account for the natural resistance of immature AML cells to anthracyclines.
Lautier
D
D
CJF INSERM 9503, Center Claudius Régaud, Toulouse, France. lautier@regaud-tlse.fnclcc.fr
Bailly
J D
JD
Demur
C
C
Herbert
J M
JM
Bousquet
C
C
Laurent
G
G
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors; D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D018952:Antigens, CD34 / Q000378:metabolism; D001386:Azides / Q000494:pharmacology; D001773:Blood Cells / Q000187:drug effects / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003078:Colchicine / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D003630:Daunorubicin / Q000378:metabolism*; D003847:Deoxyglucose / Q000494:pharmacology; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism* / Q000473:pathology; D018613:Microscopy, Confocal; D008985:Monensin / Q000494:pharmacology; D009550:Nigericin / Q000494:pharmacology; D019810:Sodium Azide; D013696:Temperature; D014407:Tumor Cells, Cultured
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000903:Antibiotics, Antineoplastic; D018952:Antigens, CD34; D001386:Azides; D003571:Cytochalasin B; D008985:Monensin; D019810:Sodium Azide; D003847:Deoxyglucose; D009550:Nigericin; D003078:Colchicine; D003630:Daunorubicin