title: gammaH2AX: A potential DNA damage response biomarker for assessing toxicological risk of tobacco products.
abstract: Differentiation among American cigarettes relies primarily on the use of proprietary tobacco blends, menthol, tobacco substitutes, paper porosity, paper additives, and filter ventilation. These characteristics substantially alter per cigarette yields of tar and nicotine in standardized protocols promulgated by government agencies. However, due to compensatory alterations in smoking behavior to sustain a preferred nicotine dose (e.g., by increasing puff frequency, inhaling more deeply, smoking more cigarettes per day, or blocking filter ventilation holes), smokers actually inhale similar amounts of tar and nicotine regardless of any cigarette variable, supporting epidemiological evidence that all brands have comparable disease risk. Consequently, it would be advantageous to develop assays that realistically compare cigarette smoke (CS)-induced genotoxicity regardless of differences in cigarette construction or smoking behavior. One significant indicator of potentially carcinogenic DNA damage is double strand breaks (DSBs), which can be monitored by measuring Ser 139 phosphorylation on histone H2AX. Previously we showed that phosphorylation of H2AX (defined as gammaH2AX) in exposed lung cells is proportional to CS dose. Thus, we proposed that gammaH2AX may be a viable biomarker for evaluating genotoxic risk of cigarettes in relation to actual nicotine/tar delivery. Here we tested this hypothesis by measuring gammaH2AX levels in A549 human lung cells exposed to CS from a range of commercial cigarettes using various smoking regimens. Results show that gammaH2AX induction, a critical event of the mammalian DNA damage response, provides an assessment of CS-induced DNA damage independent of smoking topography or cigarette type. We conclude that gammaH2AX induction shows promise as a genotoxic bioassay offering specific advantages over the traditional assays for the evaluation of conventional and nonconventional tobacco products.
authors: Vector Tobacco Inc., Public Health, 712 Fifth Avenue, 52nd Floor, New York, NY 10019, USA. talbino@vectorgroupltd.com
authors: Anthony P
authors: AP
authors: Albino
authors: Ellen D
authors: ED
authors: Jorgensen
authors: Patrick
authors: P
authors: Rainey
authors: Gene
authors: G
authors: Gillman
authors: T Jeffrey
authors: TJ
authors: Clark
authors: Diana
authors: D
authors: Gietl
authors: Hong
authors: H
authors: Zhao
authors: Frank
authors: F
authors: Traganos
authors: Zbigniew
authors: Z
authors: Darzynkiewicz
mesh_terms: D015415:Biomarkers / Q000032:analysis*; D045744:Cell Line, Tumor; D004249:DNA Damage; D006657:Histones / Q000032:analysis*; D006801:Humans; D009152:Mutagenicity Tests / Q000379:methods*; D009538:Nicotine / Q000633:toxicity*; D012306:Risk; D012907:Smoking / Q000009:adverse effects; D013638:Tars / Q000633:toxicity*
chemical_list: D015415:Biomarkers; C468783:H2AX protein, human; D006657:Histones; D013638:Tars; C024746:tobacco tar; D009538:Nicotine