The effect of zinc sulphate and zinc carnosine on genome stability and cytotoxicity in the WIL2-NS human lymphoblastoid cell line

Razinah Sharif @ Mohd Sharif, Philip Thomas, Peter Zalewski, Robin D. Graham, Michael Fenech

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Zinc (Zn) is an essential cofactor required by numerous enzymes that are essential for cell metabolism and the maintenance of DNA integrity. We investigated the effect of Zn deficiency or excess on genomic instability events and determined the optimal concentration of two Zn compounds that minimize DNA-damage events. The effects of Zn sulphate (ZnSO4) and Zn carnosine (ZnC) on cell proliferation were investigated in the WIL2-NS human lymphoblastoid cell line. DNA damage was determined by the use of both the comet assay and the cytokinesis-block micronucleus cytome (CBMN-Cyt) assay. Zn-deficient medium (0μM) was produced using Chelex treatment, and the two Zn compounds (i.e. ZnSO4 and ZnC) were tested at concentrations of 0.0, 0.4, 4.0, 16.0, 32.0 and 100.0μM. Results from an MTT assay showed that cell growth and viability were decreased in Zn-depleted cells (0μM) as well as at 32μM and 100μM for both Zn compounds (P<0.0001). DNA strand-breaks, as measured by the comet assay, were found to be increased in Zn-depleted cells compared with the other treatment groups (P<0.05). The CBMN-Cyt assay showed a significant increase in the frequency of both apoptotic and necrotic cells under Zn-deficient conditions (P<0.0001). Elevated frequencies of micronuclei (MNi), nucleoplasmic bridges (NPBs) and nuclear buds (NBuds) were induced in Zn-depleted cells (P<0.0001), whereas genome damage was reduced in supplemented cultures for both Zn compounds at 4μM and 16μM, possibly suggesting that these concentrations may be optimal for genome stability. The potential protective effect of ZnSO4 and ZnC was also investigated following exposure to 1.0Gy γ-radiation. Culture in medium containing these compounds at 4-32μM prior to irradiation displayed significantly reduced frequencies of MNi, NPBs and NBuds compared with cells maintained in 0μM medium (P<0.0001). Expression of γ-H2AX and 8-oxoguanine glycosylase measured by western blotting was increased in Zn-depleted cells. These results suggest that Zn plays important role in genomic stability and that the optimal Zn concentration-range for prevention of DNA damage and cytotoxicity in vitro lies between 4 and 16μM.

Original languageEnglish
Pages (from-to)22-33
Number of pages12
JournalMutation Research - Genetic Toxicology and Environmental Mutagenesis
Volume720
Issue number1-2
DOIs
Publication statusPublished - 28 Feb 2011

Fingerprint

Carnosine
Zinc Sulfate
Genomic Instability
Zinc
Cell Line
Zinc Compounds
DNA Damage
Micronucleus Tests
Comet Assay
Cytokinesis
DNA Breaks
Culture Media
Cell Survival

Keywords

  • Alkaline comet assay
  • DNA damage
  • Genome stability
  • Micronucleus
  • WIL2-NS cell line
  • Zinc

ASJC Scopus subject areas

  • Health, Toxicology and Mutagenesis
  • Genetics

Cite this

The effect of zinc sulphate and zinc carnosine on genome stability and cytotoxicity in the WIL2-NS human lymphoblastoid cell line. / Sharif @ Mohd Sharif, Razinah; Thomas, Philip; Zalewski, Peter; Graham, Robin D.; Fenech, Michael.

In: Mutation Research - Genetic Toxicology and Environmental Mutagenesis, Vol. 720, No. 1-2, 28.02.2011, p. 22-33.

Research output: Contribution to journalArticle

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