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• W2000352983 abstract "Human flavin-dependent monooxygenase (FMO) isoforms 1 and 3 were expressed by retroviral gene transfer in mouse C3H/10T12 cells. FMO function was determined by the sulfoxidation of p-tolylmethylsulfide (TMS). Enzyme activity ranged from 4 to 30 nmol p-tolylmethylsulfoxide (TMSO)/30 min/mg cell protein for FMO 3 clones; for FMO 1 clones, the range was 1–6 nmol TMSO/30 min/mg. Cytotoxicity in these clones after exposure to thiocarbamate compounds was assessed by clonogenic assay. Thiourea (TU), phenylthiourea (PTU), and α-naphthylthiourea (ANTU) were toxic to FMO 3 cells but not to parental and FMO 1 clones; 50% toxicity was attained at 1×10−4 M TU, 5×10−6 M PTU, and 1×10−6 M ANTU. Toxicity was observed after a minimum exposure time of 6 hr. Parental cells were resistant to toxicity for exposure times spanning the entire clonogenic assay period (10 days). Ethylene thiourea (ETU) was not toxic to FMO 3 cells, but preincubation with 1×10−3 M ETU blocked TU toxicity. Reducing GSH levels by preincubation with 1×10−5 M buthionine sulfoxime (BSO) increased TU sensitivity in FMO 3 cells from 1×10−4 to 1×10−6 M to achieve 50% toxicity. BSO also increased the sensitivity of “low expressor” FMO 3 clones to TU, but did not alter the refractoriness of either parental or FMO 1 expressing cells to TU. N-Acetylcysteine afforded modest protection to TU toxicity by shifting 50% cytotoxicity for TU from 5×10−5 to 1×10−3 M. TU mutagenicity was assayed by the development of ouabain resistance in parental and FMO 3 C3H/10T12 cells. Exposure to N-methyl-N′-nitro-N-nitrosoguanidine (MNNG, direct acting mutagen) and TU was executed with and without prior sensitization with BSO. The mutation frequency for MNNG was 76/1×106 surviving cells, whereas no mutants were observed for TU-exposed cultures. The results of this study show that, in isolation, the major human hepatic form of FMO is capable of promoting thiocarbamate toxicity. Consistent with the known reactivity of thiocarbamate intermediates with GSH, treatments that alter GSH levels also altered toxicity in either the protective or sensitizing direction. These cell lines expressing variable levels of FMO 3 and TU sensitivity should prove useful as in vitro systems for dissecting the thiocarbamate toxicity pathway." @default.
• W2000352983 created "2016-06-24" @default.
• W2000352983 creator A5069368372 @default.
• W2000352983 creator A5088332392 @default.
• W2000352983 date "2002-06-01" @default.
• W2000352983 modified "2023-10-18" @default.
• W2000352983 title "Thiourea toxicity in mouse C3H/10T12 cells expressing human flavin-dependent monooxygenase 3" @default.
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• W2000352983 doi "https://doi.org/10.1016/s0006-2952(02)00978-4" @default.
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