Intercalation of anthracyclines into living cell DNA analyzed by flow cytometry.
Anthracyclines (ANT) are used in the treatment of leukemia and other cancers. These drugs have been shown to intercalate between the strands of DNA. In the present study, we show that the amount of ANT intercalated into DNA can be determined by measuring the fluorescence resonance energy transfer (FRET) between Hoechst 33342 (H33342) and ANT bound to DNA. The transfer efficiency was found to depend on the amount of disposable ANT but was independent of the amount of H33342 bound to DNA over a wide range of H33342 concentrations. The method was adapted for flow cytometric measurement of FRET in whole living cells and was used to evaluate the degree of intercalation of daunorubicin (DAU) and idarubicine (IDA) into DAU-sensitive and DAU-resistant leukemic cell lines. ANT intercalation into DNA was affected by factors which modify the intracytoplasmic concentration of ANT, and it was shown that the action of ANT and the resistance to ANT could not be attributed solely to the intercalative effect of the drugs. The method has advantages over previously described methods and represents a useful complementary tool in studies on the mode of action of ANT and the mechanisms of chemoresistance.
Belloc
F
F
Laboratoire d'Hématologie, Hôpital Haut Lévéque, Pessac, France.
Lacombe
F
F
Dumain
P
P
Lopez
F
F
Bernard
P
P
Boisseau
M R
MR
Reifers
J
J
D001562:Benzimidazoles / Q000737:chemistry; D004247:DNA / Q000187:drug effects*; D003630:Daunorubicin / Q000737:chemistry / Q000494:pharmacology*; D004735:Energy Transfer; D005434:Flow Cytometry; D005453:Fluorescence; D006801:Humans; D015255:Idarubicin / Q000737:chemistry / Q000494:pharmacology*; D007364:Intercalating Agents / Q000737:chemistry / Q000494:pharmacology*; D015473:Leukemia, Promyelocytic, Acute / Q000473:pathology; D014407:Tumor Cells, Cultured / Q000187:drug effects
D001562:Benzimidazoles; D007364:Intercalating Agents; D004247:DNA; C017807:bisbenzimide ethoxide trihydrochloride; D015255:Idarubicin; D003630:Daunorubicin