title: Genomic instability induced by α-pinene in Chinese hamster cell line.
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.
authors.affiliation: Dipartimento di Scienze e Tecnologie Molecolari e Biomolecolari (STEMBIO), Università di Palermo, Viale delle Scienze, Parco d'Orleans, 90128 Palermo, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Catanzaro
authors.forename: Fabio
authors.initials: F
authors.lastname: Caradonna
authors.forename: Giusi
authors.initials: G
authors.lastname: Barbata
authors.forename: Marghereth
authors.initials: M
authors.lastname: Saverini
authors.forename: Maurizio
authors.initials: M
authors.lastname: Mauro
authors.forename: Giulia
authors.initials: G
authors.lastname: Sciandrello
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D000081008:Bicyclic Monoterpenes; D002478:Cells, Cultured; D002869:Chromosome Aberrations / Q000187:drug effects*; D003114:Colony-Forming Units Assay; D020552:Comet Assay; D006224:Cricetinae; D003412:Cricetulus; D004249:DNA Damage / Q000187:drug effects*; D042822:Genomic Instability / Q000187:drug effects*; D007124:Immunoenzyme Techniques; D048629:Micronuclei, Chromosome-Defective / Q000187:drug effects*; D015162:Micronucleus Tests; D039821:Monoterpenes / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000081008:Bicyclic Monoterpenes; D039821:Monoterpenes; D017382:Reactive Oxygen Species; C005451:alpha-pinene