Oxidative stress signalling: A potential mediator of tumour necrosis factor α-induced genomic instability in primary vascular endothelial cells

Mohan Natarajan, C. F. Gibbons, S. Mohan, S. Moore, M. A. Kadhim

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

Studying the potential role of tumour necrosis factor (TNF)α in the initiation of genomic instability is necessary to understand whether TNFα can serve as a signalling mediator of radiation-induced genomic instability in non-irradiated bystander cells. In this study, we examined whether TNFα could initiate processes through oxidative stress signalling that lead to DNA damage and genomic instability in primary vascular endothelium. In these cells, low linear energy transfer (LET) radiation (0.1-2 Gy) induced the secretion of TNFα into the culture medium. When added ectopically, TNFα at concentrations ranging from 0.1 ng ml-1 to 10 ng ml-1 increased (twofold to threefold) intracellular oxidative stress. Next, to examine whether TNFα induces genetic damage, cells were treated with TNFα for 5 h and analysed immediately using the single cell gel electrophoresis assay or after 3 days, 12 days and 20 days using solid stain chromosomal analysis. Cells exposed to 0.1 Gy, 1 Gy or 2 Gy or treated with 100 μM H2O2 were used as positive controls. The results showed that TNFα as low as 0.1 ng ml-1 could initiate increased DNA damage compared with untreated controls. When examined in the progeny cells after several generations, the chromosomal instability appeared to be carried over even after day 12 and day 20. The increased genetic damage is inhibited in cells that are pre-incubated with the antioxidant enzyme catalase, the antioxidant N-acetyl-L-cysteine or the metal chelator pyrrolidine dithiocarbamate. These results clearly indicate that TNFα at concentrations at which no cytotoxicity is observed could induce genetic damage through free radical generation, which could, in turn, lead to the delayed events associated with genomic instability.

Original languageEnglish (US)
Pages (from-to)S13-S22
JournalBritish Journal of Radiology
Volume80
Issue numberSPEC. ISS. 1
DOIs
StatePublished - Oct 9 2007

ASJC Scopus subject areas

  • Radiology Nuclear Medicine and imaging

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