Unknown,Transcriptomics,Genomics,Proteomics

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Genetic toxicology and toxicogenomic analysis of 3 cigarette smoke condensates in vitro reveals few differences between high and low tar brands

ABSTRACT: This experiment investigated the response of mouse pulmonary epithelial cells in culture (FE1 cell line) to 3 different brands of cigarette smoke condensate. The three brands represent a full flavour, regular and king-size light cigarettes. These vary in some key chemical consistuents. Brand #1 has the highest tar content. The cells were exposed to two doses: one at approximately 50% cytotoxicity, and the other at approximately 20%. Both doses were clastogenic. The major pathways affected included xenobiotic metabolism, cell cycle, apoptosis, oxidative stress and inflammation. Although brand #1 exhibited the most numbers of genes that were differentially expressed, there did not appear to be any major differences in the expression profiles between the brands. Although some genes did not reach statistical significance, in general there was consistency in the direction of fold changes for genes across all brands. MutaMouse FE1 cells in culture (derived from lung epithelial cells) were exposed to cigarette smoke condensate (CSC) collected from 3 brands: Brand #1 M-bM-^@M-^S Full Flavor; Brand #3 M-bM-^@M-^S Regular; Brand #5 M-bM-^@M-^S Light King Size. Experiments were performed on 5 replicates per condition (i.e., n=5 per group); one replicate was removed from 4 groups later in the study. Cells were exposed at 70% confluence in 150 mm plates to either 45 M-BM-5g/ml or 90 M-BM-5g/ml CSC alongside cells exposed to 1% DMSO (Sigma-Alderich, Canada) in 1:1 DMEM:F12 (without FBS). Cells were exposed to CSC for 6 hours and then either: (a) immediately harvested, or (b) washed in PBS and cultured for another 4 hours in fresh media. The two time points were called 6 hours or 10 hours (6 hours exposure, 4 hours recovery for the latter). Cells were harvested with TRIzol (Invitrogen) and stored at -80oC. RNA was extracted and labelled, and hybridized to Agilent 22K mouse development microarrays following the manufacturer's protocol (Agilent Technologies Inc., USA). Arrays were washed and scanned on a ScanArray Express (Perkin-Elmer Life Sciences), and data were acquired with ImaGene 5.5 (BioDiscovery Inc.). Biological samples were hybridized in Cy5, and Stratagene Universal Reference RNA was hybridized in Cy3.

ORGANISM(S): Mus musculus

SUBMITTER: Andrew Williams

PROVIDER: E-GEOD-30079 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Genetic toxicology and toxicogenomic analysis of three cigarette smoke condensates in vitro reveals few differences among full-flavor, blonde, and light products.

Yauk Carole L CL Yauk Carole L CL Williams Andrew A Buick Julie K JK Chen Guosheng G Maertens Rebecca M RM Halappanavar Sabina S White Paul A PA

Environmental and molecular mutagenesis 20120319 4

Cigarette smoking leads to various detrimental health outcomes. Tobacco companies produce different brands of cigarettes that are marketed as reduced harm tobacco products. Early examples included "light" cigarettes, which differ from regular cigarettes due to filter ventilation and/or differences in chemical constituents. In order to establish baseline similarities and differences among different tobacco brands available in Canada, the present study examined the cytotoxicity, mutagenicity, clas ...[more]

PMID: 22431010

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Project description:Like tobacco smoking, habitual marijuana smoking causes numerous adverse pulmonary effects. However, the mechanisms of action involved, especially as compared to tobacco smoke, are still unclear. To uncover putative modes of action, this study employed a toxicogenomics approach to compare the toxicological pathways perturbed following exposure to marijuana and tobacco smoke condensate in vitro. Condensates of mainstream smoke from hand-rolled tobacco and marijuana cigarettes were similarly prepared using identical smoking conditions. Murine lung epithelial cells were exposed to low, medium and high concentrations of the smoke condensates for 6 hr. RNA was extracted immediately or after a 4-hr recovery period and hybridized to mouse whole genome microarrays. Tobacco smoke condensate (TSC) exposure was associated with changes in xenobiotic metabolism, oxidative stress, inflammation, and DNA damage response. These same pathways were also significantly affected following marijuana smoke condensate (MSC) exposure. Although the effects of the condensates were largely similar, dose-response analysis indicates that the MSC is substantially more potent than TSC. In addition, steroid biosynthesis, apoptosis, and inflammation pathways were more significantly affected following MSC exposure, whereas m-phase cell cycle pathways were more significantly affected following TSC exposure. MSC exposure also appeared to elicit more severe oxidative stress than TSC exposure, which may account for the greater cytotoxicity of MSC. This study shows that in general, MSC impacts many of the same molecular processes as TSC. However, subtle pathway differences can provide insight into the differential toxicities of the two complex mixtures. Murine epithelial lung cells were exposed to tobacco smoke condensates (0, 25, 50, 90 Î¼g/ml) or marijuana smoke condensates (0, 2.5, 5, 10 Î¼g/ml) in serum-free medium for a six hour period. Following the six-hour exposure, cells were either harvested immediately or washed with phosphate-buffered saline and incubated in fresh serum-free medium for a four hour recovery period. Total RNA was extracted from the cells and hybridized against Universal Mouse Reference RNA (Agilent Technologies Canada, Inc.) to Agilent whole mouse genome microarray slides containing 44,000 transcripts. A LOWESS normalization was applied to expression results, and statistically significant genes were identified using the R library MAANOVA. Microarray results were validated by real time RT-PCR.

Dataset Information

Genetic toxicology and toxicogenomic analysis of 3 cigarette smoke condensates in vitro reveals few differences between high and low tar brands

Publications

Genetic toxicology and toxicogenomic analysis of three cigarette smoke condensates in vitro reveals few differences among full-flavor, blonde, and light products.

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