Home LiteratureArticle Details
PMID: 17169990 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Ascorbate acts as a highly potent inducer of chromate mutagenesis and clastogenesis: linkage to DNA breaks in G2 phase by mismatch repair.

Nucleic acids research ·Vol. 35 ·No. 2 ·2007-00-00 ·Pages 465-76

Reynolds M, Stoddard L, Bespalov I, Zhitkovich A

Abstract

Here we examined the role of cellular vitamin C in genotoxicity of carcinogenic chromium(VI) that requires reduction to induce DNA damage. In the presence of ascorbate (Asc), low 0.2-2 microM doses of Cr(VI) caused 10-15 times more chromosomal breakage in primary human bronchial epithelial cells or lung fibroblasts. DNA double-strand breaks (DSB) were preferentially generated in G2 phase as detected by colocalization of H2AX and 53BP1 foci in cyclin B1-expressing cells. Asc dramatically increased the formation of centromere-negative micronuclei, demonstrating that induced DSB were inefficiently repaired. DSB in G2 cells were caused by aberrant mismatch repair of Cr damage in replicated DNA, as DNA polymerase inhibitor aphidicolin and silencing of MSH2 or MLH1 by shRNA suppressed induction of H2AX and micronuclei. Cr(VI) was also up to 10 times more mutagenic in cells containing Asc. Increasing Asc concentrations generated progressively more mutations and DSB, revealing the genotoxic potential of otherwise nontoxic Cr(VI) doses. Asc amplified genotoxicity of Cr(VI) by altering the spectrum of DNA damage, as total Cr-DNA binding was unchanged and post-Cr loading of Asc exhibited no effects. Collectively, these studies demonstrated that Asc-dependent metabolism is the main source of genotoxic and mutagenic damage in Cr(VI)-exposed cells.

MeSH Terms
Ascorbic Acid/physiology Carcinogens, Environmental/metabolism,toxicity Cells, Cultured Chromium/metabolism,toxicity DNA Breaks, Double-Stranded DNA Mismatch Repair G2 Phase Histones/analysis Humans MutS Homolog 2 Protein/metabolism Mutagenesis Mutagens/metabolism,toxicity Nuclear Proteins/metabolism S Phase
Chemicals
Carcinogens, Environmental Histones Mutagens Nuclear Proteins Chromium MutS Homolog 2 Protein Ascorbic Acid
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Reynolds Mindy
Department of Pathology and Laboratory Medicine, Brown University Providence, Rhode Island 02912, USA.
Stoddard Lauren
Bespalov Ivan
Zhitkovich Anatoly
References (51)
51 references, click to expand
  1. Genotoxicity and mutagenicity of chromium(VI)/ascorbate-generated DNA adducts in human and bacterial cells.
    Biochemistry. 2003 Feb 4;42(4):1062-70 PMID: 12549927
  2. DNA damage-induced replication fork regression and processing in Escherichia coli.
    Science. 2003 Feb 14;299(5609):1064-7 PMID: 12543983
  3. Binding discrimination of MutS to a set of lesions and compound lesions (base damage and mismatch) reveals its potential role as a cisplatin-damaged DNA sensing protein.
    J Biol Chem. 2003 Jun 6;278(23):21267-75 PMID: 12654906
  4. Management of hemochromatosis. Hemochromatosis Management Working Group.
    Ann Intern Med. 1998 Dec 1;129(11):932-9 PMID: 9867745
  5. In vitro plasmid DNA cleavage by chromium(V) and -(IV) 2-hydroxycarboxylato complexes.
    Chem Res Toxicol. 1999 Apr;12(4):371-81 PMID: 10207127
  6. Megabase chromatin domains involved in DNA double-strand breaks in vivo.
    J Cell Biol. 1999 Sep 6;146(5):905-16 PMID: 10477747
  7. Hexavalent chromium and lung cancer in the chromate industry: a quantitative risk assessment.
    Risk Anal. 2004 Oct;24(5):1099-108 PMID: 15563281
  8. Importance of chromium-DNA adducts in mutagenicity and toxicity of chromium(VI).
    Chem Res Toxicol. 2005 Jan;18(1):3-11 PMID: 15651842
  9. Microsatellite instability and protein expression of the DNA mismatch repair gene, hMLH1, of lung cancer in chromate-exposed workers.
    Mol Carcinog. 2005 Mar;42(3):150-8 PMID: 15605365
  10. DNA mismatch repair.
    Annu Rev Biochem. 2005;74:681-710 PMID: 15952900
  11. Guanine-specific oxidation of double-stranded DNA by Cr(VI) and ascorbic acid forms spiroiminodihydantoin and 8-oxo-2'-deoxyguanosine.
    Chem Res Toxicol. 2005 Jul;18(7):1140-9 PMID: 16022506
  12. Mismatch repair proteins are activators of toxic responses to chromium-DNA damage.
    Mol Cell Biol. 2005 May;25(9):3596-607 PMID: 15831465
  13. Killing of chromium-damaged cells by mismatch repair and its relevance to carcinogenesis.
    Cell Cycle. 2005 Aug;4(8):1050-2 PMID: 15970712
  14. Occupational exposure to hexavalent chromium. Final rule.
    Fed Regist. 2006 Feb 28;71(39):10099-385 PMID: 16528853
  15. Ascorbate depletion mediates up-regulation of hypoxia-associated proteins by cell density and nickel.
    J Cell Biochem. 2006 Apr 1;97(5):1025-35 PMID: 16288478
  16. ATR kinase activation mediated by MutSalpha and MutLalpha in response to cytotoxic O6-methylguanine adducts.
    Mol Cell. 2006 May 19;22(4):501-10 PMID: 16713580
  17. Causes of DNA single-strand breaks during reduction of chromate by glutathione in vitro and in cells.
    Free Radic Biol Med. 2006 Jun 1;40(11):1981-92 PMID: 16716899
  18. Mechanisms in eukaryotic mismatch repair.
    J Biol Chem. 2006 Oct 13;281(41):30305-9 PMID: 16905530
  19. Lower mutagenicity but higher stability of Cr-DNA adducts formed during gradual chromate activation with ascorbate.
    Carcinogenesis. 2006 Nov;27(11):2316-21 PMID: 16714765
  20. DNA mismatch repair (MMR) mediates 6-thioguanine genotoxicity by introducing single-strand breaks to signal a G2-M arrest in MMR-proficient RKO cells.
    Clin Cancer Res. 2003 Jun;9(6):2327-34 PMID: 12796402
  21. Histone H2AX phosphorylation is dispensable for the initial recognition of DNA breaks.
    Nat Cell Biol. 2003 Jul;5(7):675-9 PMID: 12792649
  22. Report from the in vitro micronucleus assay working group.
    Mutat Res. 2003 Oct 7;540(2):153-63 PMID: 14550499
  23. Chromium inhibits transcription from polycyclic aromatic hydrocarbon-inducible promoters by blocking the release of histone deacetylase and preventing the binding of p300 to chromatin.
    J Biol Chem. 2004 Feb 6;279(6):4110-9 PMID: 14625279
  24. Dependence of the cytotoxicity of DNA-damaging agents on the mismatch repair status of human cells.
    Cancer Res. 2004 May 15;64(10):3391-4 PMID: 15150090
  25. Mechanisms and consequences of methylating agent-induced SCEs and chromosomal aberrations: a long road traveled and still a far way to go.
    Cytogenet Genome Res. 2004;104(1-4):77-86 PMID: 15162018
  26. Mismatch repair-dependent G2 checkpoint induced by low doses of SN1 type methylating agents requires the ATR kinase.
    Genes Dev. 2004 Jun 1;18(11):1331-44 PMID: 15175264
  27. ATM-mediated stabilization of hMutL DNA mismatch repair proteins augments p53 activation during DNA damage.
    Mol Cell Biol. 2004 Jul;24(14):6430-44 PMID: 15226443
  28. Human nucleotide excision repair efficiently removes chromium-DNA phosphate adducts and protects cells against chromate toxicity.
    J Biol Chem. 2004 Jul 16;279(29):30419-24 PMID: 15087443
  29. 53BP1, an activator of ATM in response to DNA damage.
    DNA Repair (Amst). 2004 Aug-Sep;3(8-9):945-52 PMID: 15279780
  30. Oxidative stress and apoptosis in metal ion-induced carcinogenesis.
    Free Radic Biol Med. 2004 Sep 1;37(5):582-93 PMID: 15288116
  31. One hundred years of chromium and cancer: a review of epidemiological evidence and selected case reports.
    Am J Ind Med. 1990;17(2):189-215 PMID: 2405656
  32. Ascorbate is the principal reductant of chromium(VI) in rat lung ultrafiltrates and cytosols, and mediates chromium-DNA binding in vitro.
    Carcinogenesis. 1992 Aug;13(8):1319-24 PMID: 1499083
  33. Mechanisms of chromium carcinogenicity and toxicity.
    Crit Rev Toxicol. 1993;23(3):255-81 PMID: 8260068
  34. Mammalian cell gene mutation assays working group report.
    Mutat Res. 1994 Jun;312(3):235-9 PMID: 7514737
  35. Reaction of chromium(VI) with ascorbate produces chromium(V), chromium(IV), and carbon-based radicals.
    Chem Res Toxicol. 1994 Mar-Apr;7(2):219-30 PMID: 8199312
  36. Efficient transport and accumulation of vitamin C in HL-60 cells depleted of glutathione.
    J Biol Chem. 1997 Apr 11;272(15):9915-21 PMID: 9092530
  37. Microprobe X-ray absorption spectroscopic determination of the oxidation state of intracellular chromium following exposure of V79 Chinese hamster lung cells to genotoxic chromium complexes.
    Chem Res Toxicol. 1997 May;10(5):533-5 PMID: 9168250
  38. PARP is important for genomic stability but dispensable in apoptosis.
    Genes Dev. 1997 Sep 15;11(18):2347-58 PMID: 9308963
  39. DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139.
    J Biol Chem. 1998 Mar 6;273(10):5858-68 PMID: 9488723
  40. Lung cancer mortality in nickel/chromium platers, 1946-95.
    Occup Environ Med. 1998 Apr;55(4):236-42 PMID: 9624277
  41. Differential effects and transport kinetics of ascorbate derivatives in leukemic cell lines.
    Anticancer Res. 1998 Jul-Aug;18(4A):2487-93 PMID: 9703897
  42. Threshold mechanisms and site specificity in chromium(VI) carcinogenesis.
    Carcinogenesis. 2000 Apr;21(4):533-41 PMID: 10753182
  43. Unmasking a killer: DNA O(6)-methylguanine and the cytotoxicity of methylating agents.
    Mutat Res. 2000 Apr;462(2-3):71-82 PMID: 10767619
  44. Lung cancer among workers in chromium chemical production.
    Am J Ind Med. 2000 Aug;38(2):115-26 PMID: 10893504
  45. Reflections on hexavalent chromium: health hazards of an industrial heavyweight.
    Environ Health Perspect. 2000 Sep;108(9):A402-7 PMID: 11017901
  46. p53 binding protein 1 (53BP1) is an early participant in the cellular response to DNA double-strand breaks.
    J Cell Biol. 2000 Dec 25;151(7):1381-90 PMID: 11134068
  47. Photoactivation of DNA thiobases as a potential novel therapeutic option.
    Curr Biol. 2001 Jul 24;11(14):1142-6 PMID: 11509240
  48. Direct oxidation of guanine and 7,8-dihydro-8-oxoguanine in DNA by a high-valent chromium complex: a possible mechanism for chromate genotoxicity.
    Chem Res Toxicol. 2001 Sep;14(9):1315-22 PMID: 11559048
  49. Carcinogenic chromium(VI) induces cross-linking of vitamin C to DNA in vitro and in human lung A549 cells.
    Biochemistry. 2002 Mar 5;41(9):3156-67 PMID: 11863455
  50. Frequent microsatellite instability in lung cancer from chromate-exposed workers.
    Mol Carcinog. 2002 Mar;33(3):172-80 PMID: 11870883
  51. Fork reversal and ssDNA accumulation at stalled replication forks owing to checkpoint defects.
    Science. 2002 Jul 26;297(5581):599-602 PMID: 12142537
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkl1069
Published
2007-00-00
Epub
2006-00-14
Pages
465-76
Language
English
Region
England
NLM ID
0411011
PMCID
PMC1802609
Subset
IM
Grants
NIEHS NIH HHS · ES008786 · United States
NIEHS NIH HHS · R01 ES008786 · United States
NIEHS NIH HHS · ES012915 · United States
NIEHS NIH HHS · R01 ES012915 · United States
NIEHS NIH HHS · ES007272 · United States
NIEHS NIH HHS · T32 ES007272 · United States
External Links
PubMed source DOI: 10.1093/nar/gkl1069 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2