Genomic instability induced by α-pinene in Chinese hamster cell line

Mutagenesis. 2012 Jul;27(4):463-9. doi: 10.1093/mutage/ges005. Epub 2012 Feb 29.

Abstract

Here, we report the effects of exposure of mammalian cells to α-pinene, a bicyclic monoterpene used in insecticides, solvents and perfumes. Morphological analysis, performed in V79-Cl3 cells exposed for 1 h to increasing concentrations (25 up to 50 μM) of α-pinene, indicated a statistically significant increase in micronucleated and multinucleated cell frequencies; apoptotic cells were seen at 40 and 50 μM. This monoterpene caused genomic instability by interfering with mitotic process; in fact, 50% of cells (versus 19% of control cells) showed irregular mitosis with multipolar or incorrectly localised spindles. Cytogenetic analysis demonstrated high-frequency hypodiploid metaphases as well as endoreduplicated cells and chromosome breaks. Clastogenic damage was prevalent over aneuploidogenic damage as demonstrated by the higher proportion of kinetochore-negative micronuclei. Alkaline comet confirmed that monoterpene exposure caused DNA lesions in a concentration-dependent manner. This damage probably arose by increased reactive oxygen species (ROS) production. In order to assess the generation of ROS, the cells were incubated with CM-H(2)DCFDA and then analysed by flow cytometry. Results demonstrated an increase in fluorescence intensity after α-pinene treatment indicating increased oxidative stress. On the whole, these findings strongly suggest that α-pinene is able to compromise genome stability preferentially through mitotic alterations and to damage DNA through ROS production.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Bicyclic Monoterpenes
  • Cells, Cultured
  • Chromosome Aberrations / drug effects*
  • Colony-Forming Units Assay
  • Comet Assay
  • Cricetinae
  • Cricetulus
  • DNA Damage / drug effects*
  • Genomic Instability / drug effects*
  • Immunoenzyme Techniques
  • Micronuclei, Chromosome-Defective / drug effects*
  • Micronucleus Tests
  • Monoterpenes / toxicity*
  • Oxidative Stress / drug effects*
  • Reactive Oxygen Species / metabolism

Substances

  • Bicyclic Monoterpenes
  • Monoterpenes
  • Reactive Oxygen Species
  • alpha-pinene