ABSTRACT
<p><b>OBJECTIVE</b>To define the differences in gene expression patterns between glycidyl methacrylate (GMA)-transformed human lung fibroblast cells (2BS cells) and controls.</p><p><b>METHODS</b>The mRNA differential display polymerase chain reaction (DD-PCR) technique was used. cDNAs were synthesized by reverse transcription and amplified by PCR using 30 primer combinations. After being screened by dot blot analysis, differentially expressed cDNAs were cloned, sequenced and confirmed by Northern blot analysis.</p><p><b>RESULTS</b>Eighteen differentially expressed cDNAs were cloned and sequenced, of which 17 were highly homologous to known genes (homology = 89%-100%) and one was an unknown gene. Northern blot analysis confirmed that eight genes encoding human zinc finger protein 217 (ZNF217), mixed-lineage kinase 3 (MLK-3), ribosomal protein (RP) L15, RPL41, RPS 16, TBX3, stanniocalcin 2 (STC2) and mouse ubiquitin conjugating enzyme (UBC), respectively, were up-regulated, and three genes including human transforming growth factor beta inducible gene (Betaig-h3), alpha-1,2-mannosidase 1A2 (MAN 1A2) gene and an unknown gene were down-regulated in the GMA-transformed cells.</p><p><b>CONCLUSION</b>Analysis of the potential function of these genes suggest that they may be possibly linked to a variety of cellular processes such as transcription, signal transduction, protein synthesis and growth, and that their differential expression could contribute to the GMA-induced neoplastic transformation.</p>
Subject(s)
Humans , Male , Air Pollutants, Occupational , Toxicity , Carcinoma, Squamous Cell , Genetics , Pathology , Cell Line, Transformed , Epoxy Compounds , Toxicity , Fibroblasts , Cell Biology , Gene Expression Profiling , Glycoproteins , Metabolism , Lung , Cell Biology , Mannosidases , Metabolism , Methacrylates , Toxicity , Mitogen-Activated Protein Kinase 3 , Metabolism , Oligonucleotide Array Sequence Analysis , Ribosomal Proteins , Metabolism , Signal Transduction , Genetics , Transforming Growth Factor beta , Metabolism , Ubiquitins , Metabolism , Zinc Fingers , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the genotoxic and nongenotoxic effects of short-term exposure to glycidyl mathacrylate (GMA) on human lung fibroblast cells (2BS cells) in vitro.</p><p><b>METHODS</b>DNA strand breakage was determined by single cell gel electrophoresis, and DNA ladder formation assay and flow cytometric analysis were carried out to detect apoptic responses of cells to GMA exposure. The HPRT gene mutation assay was used to evaluate the mutagenicity, and the effect of GMA on gap junctional intercellular communication (GJIC) in the exposed cells was examined with the scrape loading/dye transfer technique. The ability of GMA to transform 2BS cells was also tested by an in vitro cell transformation assay.</p><p><b>RESULTS</b>Exposure to GMA resulted in a dose-dependent increase in DNA strand breaks but not apoptic responses. GMA was also shown to significantly induce HPRT gene mutations and morphological transformation in 2BS cells in vitro. In contrast, GMA produced a concentration-dependent inhibition of GJIC.</p><p><b>CONCLUSIONS</b>GMA elicits both genotoxic and nongenotoxic effects on 2BS cells in vitro. The induction of DNA damage and gene mutations and inhibition of GJIC by GMA may casually contribute to GMA-induced cell transformation.</p>