Meeting Abstract
P3.64 Wednesday, Jan. 6 Ozone-induced gene dysregulation in rat and Tokay gecko lung cells: A comparative approach YAMASHITA, R.*; SAITO-REIS, C.; TAKAI, J.; DOHM, M.R.; Chaminade University, Honolulu mdohm@chaminade.edu
Ground-level ozone (O3), a significant oxidant component of urban air pollution, is linked to excess lung disease and morbidity in humans. Effects on wildlife are not known. O3 is insoluble in water and penetrates into the lung. O3 oxidizes carbohydrates, proteins, and lipids. These ozonolysis products induce dysregulation of 100s of genes in lung cells, including immune and proliferative functions. Recent studies by us on amphibians and reptiles indicate that many physiological responses are evolutionary conserved. The purpose of our study was to investigate gene expression patterns in cultured lung epithelial cells (ATCC CCL-111) from the lizard (Gekko gecko) in response to short (30 min) and repeated (30 min/day for one week) exposure to an ecologically relevant dose (0.2 parts per million O3). We used a cross species approach. qRT-PCR arrays of 84 oxidative stress, antioxidant defense, and JAK/STAT pathway genes for rats were used (SA Biosciences). First, gecko cells were exposed to O3 or clean air, then hours or days later, total RNA was extracted for cDNA, applied to arrays (cross species hybridization). Second, gecko cells were exposed to O3 or clean air, then the conditioned media was harvested and applied to cultured rat cells (CCL-149). Total RNA was extracted from the rat cells, then cDNA was made and applied to the PCR arrays. Threshold cycles were evaluated and compared to house keeping genes. Preliminary results suggest short-term O3 exposure caused transient dysregulation of 100s of genes in the vertebrate lung cells. Within hours following exposure, a subset of transcription factors and apoptosis-related genes were dysregulated; later, a distinct set of genes were dysregulated, with an emphasis on restoring redox balance. INBRE NIH Grant Number P20 RR016467-08 from the National Center for Research Resources.
