Monitoring incorporation, transformation and subcellular distribution of N-l-leucyl-doxorubicin in uterine sarcoma cells using capillary electrophoretic techniques.
Previous reports have demonstrated that N-l-leucyl-doxorubicin (LeuDox) is less toxic than its parent drug, Dox, but the underlying causes of this reduced toxicity have yet to be fully elucidated. In this study, the incorporation of LeuDox into (i) the MES-SA human uterine sarcoma cell line and (ii) its Dox resistant counterpart, MES-SA/Dx5 cell line and the subsequent transformation of LeuDox into Dox and its subcellular distribution, were investigated by micellar electrokinetic chromatography with laser-induced fluorescence detection (MEKC-LIF). In both cell lines the cellular uptakes of Dox and LeuDox were similar at equimolar doses, while the percent transformation of LeuDox into Dox in MES-SA/Dx5 cells was about twice as great as its transformation in MES-SA cells, which is beneficial for reaching Dox cytotoxic levels in this resistant cell line. When both cells lines were treated with IC(35) concentrations of either Dox and LeuDox, the intracellular Dox amounts were 6-fold higher in the resistant cell line than in the sensitive cell line, suggesting that other cellular processes play a role in the cytotoxicity of Dox in the resistant cell line. The amounts and ratios of Dox and LeuDox in four subcellular fractions of LeuDox-treated MES-SA/Dx5 cells were also investigated. The highest Dox/LeuDox ratio (i.e. 2.92) was found in the nuclear fraction, followed by the ratio in the low density organelle fraction (i.e. 1.92) that contains lysosomes, organelles in which lysosomal hydrolytic enzymes, capthesins, transform LeuDox into Dox.
Wang
Yaohua
Y
Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA.
Arriaga
Edgar A
EA
D045744:Cell Line, Tumor; D004317:Doxorubicin / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm; D019075:Electrophoresis, Capillary / Q000379:methods; D005260:Female; D006801:Humans; D011355:Prodrugs / Q000493:pharmacokinetics / Q000627:therapeutic use; D012509:Sarcoma / Q000188:drug therapy* / Q000378:metabolism; D014018:Tissue Distribution; D014594:Uterine Neoplasms / Q000188:drug therapy* / Q000378:metabolism
D011355:Prodrugs; C073409:N-leucyldoxorubicin; D004317:Doxorubicin