title: NADPH:cytochrome c (P450) reductase activates tirapazamine (SR4233) to restore hypoxic and oxic cytotoxicity in an aerobic resistant derivative of the A549 lung cancer cell line.
abstract: Tirapazamine (TPZ, SR4233, WIN 59075) is a bioreductive drug that is activated in regions of low oxygen tension to a cytotoxic radical intermediate. This labile metabolite shows high selective toxicity towards hypoxic cells, such as those found in solid tumours. Under aerobic conditions, redox cycling occurs with subsequent generation of superoxide radicals, which are also cytotoxic. NADPH:cytochrome c (P450) reductase (P450R) is a one-electron reducing enzyme that efficiently activates TPZ. Recently a derivative of the A549 non-small cell lung cancer cell line (A549c50) was generated that showed substantially reduced P450R activity compared to its parental line (Elwell et al (1997) Biochem Pharmacol 54: 249-257). Here, it is demonstrated that the A549c50 cells are markedly more resistant to TPZ under both aerobic and hypoxic conditions. In addition, these cells have a dramatically impaired ability to metabolize TPZ to its two-electron reduction product, SR4317, under hypoxic conditions when compared to wild-type cells. P450R activity in the A549c50 cells was reintroduced to similar levels as that seen in the parental A549 cells by transfection of the full-length cDNA for human P450R. These P450R over-expressing cells exhibit restored sensitivity to TPZ under both aerobic and hypoxic conditions, comparable to that found in the original parental A549 cells. Further, the ability of the transfected cells to metabolize TPZ to SR4317 under hypoxic conditions is also shown to be restored. This provides further evidence that P450R can play an important role in the activation, metabolism and toxicity of this lead bioreductive drug.
authors.affiliation: Department of Pharmacy and Pharmaceutical Sciences, University of Manchester, UK.
authors.forename: M P
authors.initials: MP
authors.lastname: Saunders
authors.forename: A V
authors.initials: AV
authors.lastname: Patterson
authors.forename: E C
authors.initials: EC
authors.lastname: Chinje
authors.forename: A L
authors.initials: AL
authors.lastname: Harris
authors.forename: I J
authors.initials: IJ
authors.lastname: Stratford
mesh_terms: D000970:Antineoplastic Agents / Q000493:pharmacokinetics*; D001711:Biotransformation; D015687:Cell Hypoxia; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism / Q000473:pathology*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009251:NADPH-Ferrihemoprotein Reductase; D010100:Oxygen / Q000378:metabolism*; D000077704:Tirapazamine; D014227:Triazines / Q000493:pharmacokinetics*; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D014227:Triazines; D000077704:Tirapazamine; D009247:NADH, NADPH Oxidoreductases; D009251:NADPH-Ferrihemoprotein Reductase; D010100:Oxygen