Zinc deficiency or excess within the physiological range increases genome instability and cytotoxicity, respectively, in human oral keratinocyte cells

Razinah Sharif @ Mohd Sharif, Philip Thomas, Peter Zalewski, Michael Fenech

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Zinc (Zn) is an essential component of Zn-finger proteins and acts as a cofactor for enzymes required for cellular metabolism and in the maintenance of DNA integrity. The study investigated the genotoxic and cytotoxic effects of Zn deficiency or excess in a primary human oral keratinocyte cell line and determined the optimal concentration of two Zn compounds (Zn Sulphate (ZnSO 4) and Zn Carnosine (ZnC)) to minimiseDNAdamage. Zn-deficient medium (0 μM) was produced using Chelex treatment, and the two Zn compounds ZnSO 4 and ZnC were tested at concentrations of 0.0, 0.4, 4.0, 16.0, 32.0 and 100.0 μM. Cell viability was decreased in Zndepleted cells (0 μM) as well as at 32 μM and 100 μM for both Zn compounds (P<0.0001) as measured via the MTT assay. 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 Cytokinesis Block Micronucleus Cytome assay showed a significant increase in the frequency of both apoptotic and necrotic cells under Zn-deficient conditions (P<0.05). Furthermore, elevated frequencies of micronuclei (MNi), nucleoplasmic bridges (NPBs) and nuclear buds (NBuds) were observed at 0 and 0.4 μM Zn, whereas these biomarkers were minimised for both Zn compounds at 4 and 16 μM Zn (P<0.05), suggesting these concentrations are optimal to maintain genome stability. Expression of PARP, p53 and OGG1 measured by western blotting was increased in Zn-depleted cells indicating that DNA repair mechanisms are activated. These results suggest that maintaining Zn concentrations within the range of 4-16 μM is essential for DNA damage prevention in cultured human oral keratinocytes.

Original languageEnglish
Pages (from-to)139-154
Number of pages16
JournalGenes and Nutrition
Volume7
Issue number2
DOIs
Publication statusPublished - Apr 2012

Fingerprint

Genomic Instability
Keratinocytes
Zinc
Zinc Compounds
Carnosine
Zinc Sulfate
Micronucleus Tests
DNA Breaks
Comet Assay
Cytokinesis
Coenzymes
Zinc Fingers
DNA Repair
DNA Damage
Cell Survival
Biomarkers
Western Blotting
Maintenance
Cell Line

Keywords

  • Cytotoxicity
  • DNA damage
  • Genomic stability
  • Human oral keratinocytes
  • Micronuclei
  • Zinc

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Genetics

Cite this

Zinc deficiency or excess within the physiological range increases genome instability and cytotoxicity, respectively, in human oral keratinocyte cells. / Sharif @ Mohd Sharif, Razinah; Thomas, Philip; Zalewski, Peter; Fenech, Michael.

In: Genes and Nutrition, Vol. 7, No. 2, 04.2012, p. 139-154.

Research output: Contribution to journalArticle

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