Archives of Toxicology

, Volume 78, Issue 1, pp 7–15 | Cite as

Vanadyl sulfate can differentially damage DNA in human lymphocytes and HeLa cells

  • Katarzyna WozniakEmail author
  • Janusz Blasiak
Inorganic Compounds


Using the comet assay, we showed that vanadyl sulfate induced DNA damage in human normal lymphocytes and in HeLa cells. Vanadyl at 0.5 and 1 mM produced DNA single- and double-strand breaks (SSBs and DSBs) in lymphocytes, whereas in HeLa cells we observed only SSBs. Post-treatment of vanadyl-damaged DNA from lymphocytes with formamidopyrimidine-DNA glycosylase (Fpg), an enzyme recognizing oxidized purines, gave rise to a significant increase in the extent of DNA damage. A similar effect was observed in HeLa cells, but, using endonuclease III, we also detected oxidized pyrimidines in DNA of these cells. There were no differences in the extent of DNA damage in the lymphocytes and HeLa cells in the pH >13 and pH 12.1 conditions of the comet assay, which indicates that strand breaks, and not alkali-labile sites, contributed to the measured DNA damage. Study of DNA repair, determined in the comet assay as an ability of cells to decrease of DNA damage, revealed that HeLa cells retained the ability to repair vanadyl-damaged DNA induced at a ten-fold higher concentration than that in lymphocytes. Incubation of the cells with nitrone spin traps DMPO, POBN and PBN decreased the extent of DNA damage, which might follow from the production of free radicals by vanadyl sulfate. The presence of vitamins A, C or E caused an increase of DNA damage in HeLa cells whereas in lymphocytes such an increase was observed only for vitamin C. Our data indicate that vanadyl sulfate can be genotoxic for normal and cancer cells. It seems to have a higher genotoxic potential for cancer cells than for normal lymphocytes. Vitamins A, C and E can increase this potential.


Vanadyl sulfate Human lymphocytes HeLa cells Comet assay Spin trapping Vitamins A, C and E 



We would like to thank Magdalena Jaskula for her technical assistance. The work was supported by the grant 505/449 (K.W.) and 505/450 (J.B.) from the University of Lodz, Poland.


  1. Ashby JA, Tinwell H, Lefevre PA, Browne MA (1995) The single cell gel electrophoresis assay for induced DNA damage (comet assay): measurement of tail length and moment. Mutagenesis 10:85–90PubMedGoogle Scholar
  2. Barbeau B, Bernier R, Dumais N, Briand G, Olivier M, Faure R, Posner BI Tremblay M (1997) Activation of HIV-1 long terminal repeat transcription and virus replication via NF-κB-dependent and -independent pathways by potent phosphotyrosine phosphatase inhibitors, the peroxovanadium compounds. J Biol Chem 272:12968–12977CrossRefPubMedGoogle Scholar
  3. Bevan AP, Drake PG, Yale JF, Shaver A, Posner BI (1995) Peroxovanadium compounds: biological actions and mechanism of insulin-mimesis. Mol Cell Biochem 153:49–58PubMedGoogle Scholar
  4. Blasiak J, Kowalik J (2000) A comparison of the in vitro genotoxicity of tri- and hexavalent chromium. Mutat Res 469:135–145CrossRefPubMedGoogle Scholar
  5. Carr A, Frei B (1999) Does Vitamin C act as a pro-oxidant under physiological conditions? FASEB J 13:1007–1012PubMedGoogle Scholar
  6. Catani MV, Costanzo A, Savini I, Levrero M, De Laurenzi V, Wang JYJ, Melino G, Avigliano L (2002) Ascorbate up-regulates MLH1 (Mut L homologue-1) and p73: implications for the cellular response to DNA damage. Biochem J 364:441–447CrossRefPubMedGoogle Scholar
  7. Collins AR, Duthie SJ, Dobson VL (1993) Direct enzymatic detection of endogenous base damage in human lymphocyte DNA. Carcinogenesis 14:1733–1735PubMedGoogle Scholar
  8. Cortizo AM, Salice VC, Vescina CM, Etcheverry SB (1997) Proliferative and morfological changes induced by vanadium compounds on Swiss 3T3 fibroblasts. Biometals 10:127–133PubMedGoogle Scholar
  9. Cusi K, Cukier S, DeFronzo RA, Torres M, Puchulu FM, Pereira Redondo JC (2001) Vanadyl sulfate improves hepatic and muscle insulin sensitivity in type 2 diabetes. J Clin Endocrinol Metab 86:1410–1417PubMedGoogle Scholar
  10. Dong Y, Narla RK, Sudbeck E, Uckun FM (2000) Synthesis, X-ray structure, and anti-leukemic activity of oxovanadium(IV) complexes. J Inorg Biochem 78:321–330CrossRefPubMedGoogle Scholar
  11. Evans MD, Podmore ID, Daly GJ, Perrett D, Lunec J, Herbert KE (1995) Detection of purine lesions in cellular DNA using single cell gel electrophoresis with Fpg protein. Biochem Soc Trans 23:434SPubMedGoogle Scholar
  12. Gallo-Hendrikx E, Murray SA, Vonderhaar BK, Xiao Z-XJ (2001) Vanadate disrupts mammary gland development in whole organ culture. Dev Dyn 222:354–367CrossRefPubMedGoogle Scholar
  13. Hanauske U, Hanauske AR, Marshall MH, Muggia VA, Von Hoff DD (1987) Biphasic effect of vanadium salts on in vitro tumor colony growth. Int J Cell Cloning 5:170–178PubMedGoogle Scholar
  14. Heffetz D, Bushkin H, Dror R, Zick Y (1990) The insulinomimetic agents H2O2 and vanadate stimulate protein tyrosine phosphorylation in intact cells. J Biol Chem 265:2896–2902PubMedGoogle Scholar
  15. Hori C, Oka T (1980) Vanadate enhances the stimulatory action of insulin on DNA synthesis in cultured mouse mammary gland. Biochim Biophys Acta 610:235–240CrossRefPubMedGoogle Scholar
  16. Huang C, Ding M, Li J, Leonard SS, Rojanasakul Y, Castranova V, Vallyathan V, Ju G, Shi X (2001) Vanadium-induced nuclear factor of activated T cells activation through hydrogen peroxide. J Biol Chem 276:22397–22403CrossRefPubMedGoogle Scholar
  17. Ivancsits S, Pilger A, Diem E, Schaffer A, Rudiger HW (2002) Vanadate induces DNA strand breaks in cultured human fibroblasts at doses relevant to occupational exposure. Mutat Res 519:25–35CrossRefPubMedGoogle Scholar
  18. Jaspers I, Samet JM, Erzurum S, Reed W (2000) Vanadium-induced κB-dependent transcription depends upon peroxide-induced activation of the p38 mitogen-activated protein kinase. Am J Respir Cell Mol Biol 23:95–102PubMedGoogle Scholar
  19. Klaude M, Eriksson S, Nygren J, Ahnstrom G (1996) The comet assay: mechanisms and technical considerations. Mutat Res 363:89–96PubMedGoogle Scholar
  20. Krokan HE, Standal R, Slupphaug G (1997) DNA glycosylases in the base excision repair. Biochem J 325:1-16PubMedGoogle Scholar
  21. McNeill JH, Yuen VG, Dai S, Orvig C (1995) Increased potency of vanadium using organic ligands. Mol Cell Biochem 153:175–180PubMedGoogle Scholar
  22. Morinville A, Maysinger D, Shaver A (1998) From vanadis to atropos: vanadium compounds as pharmacological tools in cell death signalling. Trends Pharmacol Sci 19:452–460CrossRefPubMedGoogle Scholar
  23. Murata M, Kawanishi S (2000) Oxidative DNA damage by vitamin A and its derivative via superoxide generation. J Biol Chem 275:2003–2008CrossRefPubMedGoogle Scholar
  24. Rojas E, Valverde M, Herrera LA, Altamirano-Lozano M, Ostrosky-Wegman P (1996) Genotoxicity of vanadium pentoxide evaluate by the single cell gel electrophoresis assay in human lymphocytes. Mutat Res 359:77–84CrossRefPubMedGoogle Scholar
  25. Rojas E, Herrera LA, Poirier LA, Ostrosky-Wegman P (1999) Are metals dietary carcinogens? Mutat Res 443:157–181PubMedGoogle Scholar
  26. Ruff SJ, Chen K, Cohen S (1997) Peroxovanadate induces tyrosine phosphorylation of multiple signaling proteins in mouse liver and kidney. J Biol Chem 272:1263–1267CrossRefPubMedGoogle Scholar
  27. Sakurai H (2002) A new concept: the use of vanadium complexes in the treatment of diabetes mellitus. Chem Rec 2:237–248CrossRefPubMedGoogle Scholar
  28. Sakurai H, Nakai M, Miki T, Tsuchiya K, Takada J, Matsushita R (1992) DNA cleavage by hydroxyl radicals generated in a vanadyl ion-hydrogen peroxide system. Biochem Biophys Res Commun 189:1090–1095PubMedGoogle Scholar
  29. Sakurai H, Tamura H, Okatani K (1995) Mechanism for a new antitumor vanadium complex: hydroxyl radical-dependent DNA cleavage by 1,10-phenanthroline-vanadyl complex in the presence of hydrogen peroxide. Biochem Biophys Res Commun 206:133–137CrossRefPubMedGoogle Scholar
  30. Sakurai H, Kojima Y, Yoshikawa Y, Kawabe K, Yasui H (2002) Antidiabetic vanadium(IV) and zinc(II) complexes. Coordin Chem Rev 226:187–198CrossRefGoogle Scholar
  31. Sheu CW, Rodriguez I, Lee JK (1992) Proliferation and morphological transformation of BALB/3T3 cells by a prolonged treatment with sodium orthovanadate. Food Chem Toxicol 30:307–311CrossRefPubMedGoogle Scholar
  32. Shi X, Wang P, Jiang H, Mao Y, Ahmed N, Dalal N (1996) Vanadium(IV) causes 2′-deoxyguanosine hydroxylation and deoxyribonucleic acid damage via free radical reactions. Ann Clin Lab Sci 26:39–49PubMedGoogle Scholar
  33. Singh NP, Stephens RE (1997) Microgel electrophoresis: sensitivity, mechanisms and DNA electrostretching. Mutat Res 383:167–175PubMedGoogle Scholar
  34. Singh NP, McCoy T, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–192PubMedGoogle Scholar
  35. Smith JB (1983) Vanadium ions stimulate DNA synthesis in Swiss mouse 3T3 and 3T6. Proc Natl Acad Sci USA 80:6162–6166PubMedGoogle Scholar
  36. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Miyamae Y, Rojas E, Ryu J-C, Sasaki Y-F (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35:206–221PubMedGoogle Scholar
  37. Wozniak K, Blasiak J (2002) Free radicals-mediated induction of oxidized DNA bases and DNA-protein cross-links by nickel chloride. Mutat Res 514:233–243CrossRefPubMedGoogle Scholar
  38. Wozniak K, Blasiak J (2003) In vitro genotoxicity of lead acetate: induction of single and double DNA strand breaks and DNA-protein cross-links. Mutat Res 535:127–139PubMedGoogle Scholar
  39. Yamashita N, Murata M, Inoue S, Burkitt MJ, Milne L, Kawanishi S (1998) α-Tocopherol induces oxidative damage to DNA in the presence of copper(II) ions. Chem Res Toxicol 11:855–862CrossRefPubMedGoogle Scholar
  40. Zhang Z, Huang C, Li J, Leoanrd SS, Lanciotti R, Butterworth L, Shi X (2001) Vanadate-induced cell growth regulation and the role of reactive oxygen species. Arch Biochem Biophys 392:311–320CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Department of Molecular GeneticsUniversity of LodzLodzPoland

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