Reversal of mitomycin C resistance by overexpression of bioreductive enzymes in Chinese hamster ovary cells.
The clinical utility of antineoplastic agents is limited by the development of drug resistance by tumors. Mitomycin C (MC) is a bacterial product that must be enzymatically reduced to exert anticancer activity. We have demonstrated that expression of the bacterial MC resistance-associated (MCRA) protein in Chinese hamster ovary (CHO) cells confers profound resistance to this antibiotic under aerobic conditions, but not under hypoxia. MCRA produces resistance to MC by redox cycling of the activated hydroquinone intermediate back to the prodrug form. A CHO cell line developed by stepwise exposure to increasing concentrations of MC likewise expressed high level resistance to MC in air, but not under hypoxia. The overexpression of DT-diaphorase and NADPH:cytochrome c (P-450) reductase, two enzymes known to activate MC, restored sensitivity to MC in both MCRA-transfected and drug-selected cell lines. The level of sensitization was proportional to the quantity of enzyme activity expressed, supporting the concept that the levels of these two activating enzymes are important for sensitivity to MC. The findings of resistance to MC in air but not under hypoxic conditions and of restoration of sensitivity to MC by increasing levels of DT-diaphorase activity, properties not adequately explained by other resistance mechanisms (i.e., decreases in MC activation, repair of DNA lesions, and/or drug efflux), support the hypothesis that a functional mammalian homologue of MCRA may be involved in producing resistance to MC.
Baumann
R P
RP
Department of Pharmacology, Cancer Center, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
Hodnick
W F
WF
Seow
H A
HA
Belcourt
M F
MF
Rockwell
S
S
Sherman
D H
DH
Sartorelli
A C
AC
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000493:pharmacokinetics / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics; D001711:Biotransformation; D016466:CHO Cells / Q000187:drug effects / Q000201:enzymology; D015687:Cell Hypoxia / Q000502:physiology; D006224:Cricetinae; D019008:Drug Resistance, Neoplasm; D016685:Mitomycin / Q000493:pharmacokinetics / Q000494:pharmacology*; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D009251:NADPH-Ferrihemoprotein Reductase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D010088:Oxidoreductases; D010100:Oxygen / Q000378:metabolism; D014162:Transfection
D000903:Antibiotics, Antineoplastic; D001426:Bacterial Proteins; D016685:Mitomycin; D010088:Oxidoreductases; C088145:mcrA protein, Streptomyces lavendulae; D009251:NADPH-Ferrihemoprotein Reductase; D016660:NAD(P)H Dehydrogenase (Quinone); D010100:Oxygen