Quantitative study of doxorubicin in living cell nuclei by microspectrofluorometry.
Doxorubicin-DNA association has been studied by quantitative microspectrofluorometry. Fluorescence emission spectra from a microvolume of single living cell nuclei treated with doxorubicin have been analyzed in terms of difference in spectral shape and fluorescence yield between free and DNA-bound drug. Contribution of each spectral component to the total signal was calculated by least-squares linear regression. With this method of analysis, total drug concentration has been determined with an error of less than 10%. Moreover, the uptake into the nucleus has been studied in a non destructive way, avoiding use of 14C-labelled drug. Kinetic studies of drug accumulation into the nuclei were conducted on sensitive and resistant cells.
Gigli
M
M
Laboratoire de Spectroscopie Biomolèculaire, Faculté de Pharmacie, Reims, France.
Doglia
S M
SM
Millot
J M
JM
Valentini
L
L
Manfait
M
M
D002467:Cell Nucleus / Q000032:analysis*; D004273:DNA, Neoplasm / Q000032:analysis; D004317:Doxorubicin / Q000032:analysis*; D004351:Drug Resistance; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology; D008832:Microchemistry; D013050:Spectrometry, Fluorescence; D014407:Tumor Cells, Cultured / Q000032:analysis / Q000187:drug effects
D004273:DNA, Neoplasm; D004317:Doxorubicin