Differential formation of hydroxyl radicals by adriamycin in sensitive and resistant MCF-7 human breast tumor cells: implications for the mechanism of action.
Adriamycin-stimulated formation of .OH in sensitive and resistant subline of human breast tumor cells (MCF-7) has been examined by electron spin resonance spectroscopy. It was shown that adriamycin significantly stimulated the formation of .OH spin adducts [5,5-dimethyl-1-pyrroline N-oxide (DMPO)-OH] in the sensitive cells but not in the resistant cells. By use of spin-broadening techniques and inhibition of .OH with high molecular weight poly(ethylene glycol), which does not enter intact cells, it was shown that 60-65% of adriamycin-induced .OH were located extracellularly and were metal ion dependent since they were decreased in the presence of desferal. Furthermore, superoxide dismutase and catalase, enzymes that detoxify superoxide and hydrogen peroxide, also significantly inhibited adriamycin-induced .OH formation and protected against the cytotoxicity of adriamycin. The differential .OH formation in these two cell lines is not due to diminished activities of flavin-dependent activating enzymes nor decreased accumulation of the drug in the cells but appears to be related to enhanced activities of detoxifying enzymes, particularly, glutathione peroxidases in the resistant cells.
Sinha
B K
BK
Clinical Pharmacology Branch, National Cancer Institute, Bethesda, Maryland 20892.
Katki
A G
AG
Batist
G
G
Cowan
K H
KH
Myers
C E
CE
D001943:Breast Neoplasms / Q000188:drug therapy* / Q000201:enzymology; D002374:Catalase / Q000502:physiology; D002460:Cell Line; D003114:Colony-Forming Units Assay; D003497:Cyclic N-Oxides / Q000494:pharmacology; D003676:Deferoxamine / Q000494:pharmacology; D004317:Doxorubicin / Q000494:pharmacology*; D004351:Drug Resistance; D004578:Electron Spin Resonance Spectroscopy; D005260:Female; D005609:Free Radicals; D005979:Glutathione Peroxidase / Q000502:physiology; D006801:Humans; D006878:Hydroxides / Q000096:biosynthesis; D017665:Hydroxyl Radical; D010100:Oxygen / Q000378:metabolism; D013482:Superoxide Dismutase / Q000502:physiology; D014410:Tumor Stem Cell Assay
C025634:5,5-dimethylpyrrolidin-2-one-1-oxide; D003497:Cyclic N-Oxides; D005609:Free Radicals; D006878:Hydroxides; D017665:Hydroxyl Radical; D004317:Doxorubicin; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D003676:Deferoxamine; D010100:Oxygen