Influence of N-methylformamide on the intracellular transport of doxorubicin.
The polar solvent N-methylformamide proved to be capable of enhancing the cytotoxic potential of various antitumoral compounds, both in vitro and in vivo. In many cases, this ability depended on the sequence of treatment, and the enhancement of the cytotoxic effect occurred only when N-methylformamide administration succeeded anticancer drug treatment. The results obtained in the present study indicate that N-methylformamide interferes with the mechanisms of intracellular transport and efflux of the antitumoral drug doxorubicin. In particular, laser scanning confocal microscopy observations performed on melanoma cells (M14) after N-methylformamide administration revealed evident alterations of the microtubular network, including numerous interruptions of the microtubules. Moreover, when doxorubicin-treated cells were recovered in the presence of the polar solvent, the normal efflux of the anthracyclinic antibiotic appeared to be hampered, and the drug was localized mainly in well delimited perinuclear regions. Double staining experiments demonstrated the colocalization of the doxorubicin molecules and the WGA-stained regions as well as a close structural relationship between them and the microtubule system. These results indicate that N-methylformamide interferes with the doxorubicin transport inducing a damage in the microtubular network and the consequent persistence and entrapment of the drug in the regions likely occupied by the Golgi apparatus of tumor cells. This finding could account for the chemosensitizing properties exerted by N-methylformamide.
Calcabrini
A
A
Laboratorio di Ultrastrutture, Istituto Superiore di Sanità, Rome/Italy.
Villa
A M
AM
Molinari
A
A
Doglia
S M
SM
Arancia
G
G
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D004317:Doxorubicin / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods; D005559:Formamides / Q000494:pharmacology*; D018613:Microscopy, Confocal; D008870:Microtubules / Q000187:drug effects; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D005559:Formamides; D004317:Doxorubicin; C002950:methylformamide