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Increased mutant frequencies in the HPRT gene locus of leukemia HL-60 cells treated with succinylacetone

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Abstract

Succinyl acetone (SA) was initially identified in the urine of patients with tyrosinemia type I, an autosomally recessive inherited disease. SA has been used to downregulate the activity of myeloperoxidase (MPO) through its specific inhibition of heme biosynthesis and to investigate the biological properties of MPO in the human myeloid leukemic (HL-60) cell line. The goal of this study is to evaluate the mutagenic potential of SA by determining the frequencies of somatic mutations in the hypoxanthine-guanine phosphoribosyl transferase (HPRT) reporter gene in HL-60 cells following treatment with the chemical. Treatments of HL-60 cells with 500 μmol/L SA for 72 h, a condition generally used to inhibit the MPO activity, resulted in a significantly increased HPRT mutant frequency (HPRT-Mf), compared with the control of untreated cells (47.25 × 10-6 versus 7.5 × 10-6, respectively, p <0.01). Treatment of the cells with lower doses of SA also led to an increase in HPRT-Mf but this was significant only with 200 μmol/L (28.67 × 10-6, p<0.05) and not with doses lower than 100 μmol/L (p0.05), compared with the control of untreated cells (7.5 × 10-6). These data show a dose–response increase in HPRT-Mf in HL-60 cells treated with SA, suggesting that this chemical causes mutations in the HPRT locus in these cells either directly or indirectly through its inhibition of the MPO activity.

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Abbreviations

6-TG:

6-thioguanine

ALA-D:

5-aminolevulinate dehydratase

BPDE:

benzo[a]pyrene 7,8-diol-9,10-epoxide

DMSO:

dimethyl sulfoxide

FAA:

fumarylacetoacetate

HPRT:

hypoxanthine-guanine phosphoribosyl transferase

HPRT-Mf:

HPRT mutant frequency

MPO:

myeloperoxidase

SA:

succinylacetone

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Authors and Affiliations

  1. Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

    K.-C. Zheng, V. E. Kagan & P. Keohavong

  2. Department of Pharmacology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

    J. C. Yalowich

  3. University of Pittsburgh Cancer Institute, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

    J. C. Yalowich, V. E. Kagan & P. Keohavong

  4. Department of Environmental and Preventive Medicine, School of Medicine, University of the Ryukyus, Okinawa, Japan

    K.-C. Zheng

  5. Department of Environmental and Occupational Health, Graduate School of Public Health, University of Pittsburgh, 100 Technology Drive, Bridgeside Point, Suite 350, Pittsburgh, PA, 15219–3130, USA

    P. Keohavong

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  1. K.-C. Zheng
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  2. J. C. Yalowich
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  3. V. E. Kagan
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  4. P. Keohavong
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Correspondence to P. Keohavong.

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Zheng, KC., Yalowich, J.C., Kagan, V.E. et al. Increased mutant frequencies in the HPRT gene locus of leukemia HL-60 cells treated with succinylacetone. Cell Biol Toxicol 22, 361–370 (2006). https://doi.org/10.1007/s10565-006-0171-4

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