Transferrin as a drug carrier: Cytotoxicity, cellular uptake and transport kinetics of doxorubicin transferrin conjugate in the human leukemia cells

Abstract

Leukemias are one of most common malignancies worldwide. There is a substantial need for new chemotherapeutic drugs effective against this cancer. Doxorubicin (DOX), used for treatment of leukemias and solid tumors, is poorly efficacious when it is administered systemically at conventional doses. Therefore, several strategies have been developed to reduce the side effects of this anthracycline treatment. In this study we compared the effect of DOX and doxorubicin-transferrin conjugate (DOX-TRF) on human leukemia cell lines: chronic erythromyeloblastoid leukemia (K562), sensitive and resistant (K562/DOX) to doxorubicin, and acute lymphoblastic leukemia (CCRF-CEM). Experiments were also carried out on normal cells, peripheral blood mononuclear cells (PBMC). We analyzed the chemical structure of DOX-TRF conjugate by using mass spectroscopy. The in vitro growth-inhibition assay XTT, indicated that DOX-TRF is more cytotoxic for leukemia cells sensitive and resistant to doxorubicin and significantly less sensitive to normal cells compared to DOX alone. During the assessment of intracellular DOX-TRF accumulation it was confirmed that the tested malignant cells were able to retain the examined conjugate for longer periods of time than normal lymphocytes. Comparison of kinetic parameters showed that the rate of DOX-TRF efflux was also slower in the tested cells than free DOX. The results presented here should contribute to the understanding of the differences in antitumor activities of the DOX-TRF conjugate and free drug.

Keywords: CCRF-CEM; Cytotoxicity; DOX; DOX–TRF; Doxorubicin (DOX); Doxorubicin–transferrin conjugate (DOX–TRF); E(t)(=60); IMS; Intracellular drug accumulation; Ion Mobility Mass Spectrometry; K562; Leukemia cells; PBMC; TRF; U(t)(=60); V(in); V(out); acute lymphoblastic leukemia cells; chronic erythromyeloblastoid leukemia cells; collisional cross section; doxorubicin; doxorubicin–transferrin conjugate; drift times; drug removed by cells within 60min; drug taken up by cells within 60min; efflux rate; efflux rate constant; influx rate; influx rate constant; k(in); k(out); peripheral blood mononuclear cells; tD; transferrin; Ω.