title: Effect of NQO1 induction on the antitumor activity of RH1 in human tumors in vitro and in vivo.
abstract: NQO1 is a reductive enzyme that is important for the activation of many bioreductive agents and is a target for an enzyme-directed approach to cancer therapy. It can be selectively induced in many tumor types by a number of compounds including dimethyl fumarate and sulforaphane. Mitomycin C is a bioreductive agent that is used clinically for treatment of solid tumors. RH1 (2,5-diaziridinyl-3-(hydroxymethyl)- 6-methyl-1,4-benzoquinone) is a new bioreductive agent currently in clinical trials. We have shown previously that induction of NQO1 can enhance the antitumor activity of mitomycin C in tumor cells in vitro and in vivo. As RH1 is activated selectively by NQO1 while mitomycin C is activated by many reductive enzymes, we investigated whether induction of NQO1 would produce a greater enhancement of the antitumor activity of RH1 compared with mitomycin C. HCT116 human colon cancer cells and T47D human breast cancer cells were incubated with or without dimethyl fumarate or sulforaphane followed by mitomycin C or RH1 treatment, and cytotoxic activity was measured by a clonogenic (HCT116) or MTT assay (T47D). Dimethyl fumarate and sulforaphane treatment increased NQO1 activity by 1.4- to 2.8-fold and resulted in a significant enhancement of the antitumor activity of mitomycin C, but not of RH1. This appeared to be due to the presence of a sufficient constitutive level of NQO1 activity in the tumor cells to fully activate the RH1. Mice were implanted with HL60 human promyelocytic leukemia cells, which have low levels of NQO1 activity. The mice were fed control or dimethyl fumarate-containing diet and were treated with RH1. NQO1 activity in the tumors increased but RH1 produced no antitumor activity in mice fed control or dimethyl fumarate diet. This is consistent with a narrow window of NQO1 activity between no RH1 activation and maximum RH1 activation. This study suggests that selective induction of NQO1 in tumor cells is not likely to be an effective strategy for enhancing the antitumor activity of RH1. In addition, we found that RH1 treatment produced significant leukopenia in mice that may be of concern in the clinic. These results suggest that the ease of reduction of RH1 by NQO1 makes it a poor candidate for an enzyme-directed approach to cancer therapy.
authors.affiliation: Manitoba Institute of Cell Biology, CancerCare Manitoba, University of Manitoba, 675 McDermot Avenue, Winnipeg, Manitoba, R3E 0V9, Canada.
authors.forename: Tyler
authors.initials: T
authors.lastname: Digby
authors.forename: Marsha K
authors.initials: MK
authors.lastname: Leith
authors.forename: James A
authors.initials: JA
authors.lastname: Thliveris
authors.forename: Asher
authors.initials: A
authors.lastname: Begleiter
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000633:toxicity; D001388:Aziridines / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D016227:Benzoquinones / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D045744:Cell Line, Tumor / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004790:Enzyme Induction; D005260:Female; D005562:Formazans / Q000378:metabolism; D007970:Leukopenia / Q000139:chemically induced / Q000473:pathology; D051379:Mice; D008815:Mice, Inbred Strains; D008819:Mice, Nude; D016685:Mitomycin / Q000378:metabolism / Q000494:pharmacology; D016660:NAD(P)H Dehydrogenase (Quinone); D009252:NADPH Dehydrogenase / Q000096:biosynthesis*; D009369:Neoplasms / Q000188:drug therapy* / Q000201:enzymology; D013778:Tetrazolium Salts / Q000378:metabolism; D018675:Toxicity Tests; D023041:Xenograft Model Antitumor Assays
chemical_list: C117776:2,5-diaziridinyl-3-(hydroxymethyl)-6-methyl-1,4-benzoquinone; D000970:Antineoplastic Agents; D001388:Aziridines; D016227:Benzoquinones; D005562:Formazans; D013778:Tetrazolium Salts; C079782:MTT formazan; D016685:Mitomycin; D016660:NAD(P)H Dehydrogenase (Quinone); C491732:Nqo1 protein, mouse; D009252:NADPH Dehydrogenase