Gene expression profiling in lung tissues from rats exposed to formaldehyde

Formaldehyde is a ubiquitous toxic organic compound recently classified as a carcinogen by the International Agency for Research on Cancer and one of the major factors causing sick building syndrome. In this study, we have investigated the effects of formaldehyde on mRNA expression in rat lung tissues by applying genomics. Rats were exposed to ambient air and two different concentrations of formaldehyde (0, 5, 10 ppm) for 2 weeks at 6 h/day and 5 days/week in an inhalation chamber. Malondialdehyde (MDA) assay and carbonyl spectrometric assay were conducted to determine lipid peroxidation and protein oxidation levels and Comet assays were used for genotoxicity evaluation. Level of MDA, carbonyl insertion and DNA damage in the lungs of rats exposed to FA were found to be dose dependently increased. Gene expression was evaluated by using a bio-array hybridization analysis. A total of 21 (2 up- and 19 down-regulated) genes were identified as biomarkers for formaldehyde effects. Several differentiated gene groups were found. Genes involved in apoptosis, immunity, metabolism, signal transduction, transportation, coagulation and oncogenesis were found to be up- and down-regulated. Among these genes, the mRNA expressions of cytochrome P450, hydroxymethylbilane synthase, glutathione reductase, carbonic anhydrase 2, natriuretic peptide receptor 3, lysosomal associated protein transmembrane 5, regulator of G-protein signaling 3, olfactomedin related ER localized protein, and poly (ADP-ribose) polymerase-1 were confirmed by quantitative RT-PCR analysis. In summary, the MDA lipid peroxidation and the carbonyl protein oxidation assays showed that cytotoxic effects increased with increasing formaldehyde levels. Genomic analysis showed that 21 genes were up- or down-regulated. Of these genes, nine were confirmed by quantitative RT-PCR and could be potential biomarkers for human diseases associated with formaldehyde exposure.

[1]  M. E. Meek,et al.  Inhaled Formaldehyde: Exposure Estimation, Hazard Characterization, and Exposure-Response Analysis , 2003, Journal of toxicology and environmental health. Part B, Critical reviews.

[2]  G. Heuser,et al.  Evidence for formaldehyde antibodies and altered cellular immunity in subjects exposed to formaldehyde in mobile homes. , 1987, Archives of environmental health.

[3]  R. M. Adibhatla,et al.  Serial Review: Redox-regulated Phospholipase Signal Transduction Serial Review Editors: Phospholipase a 2 , Reactive Oxygen Species, and Lipid Peroxidation in Cerebral Ischemia , 2005 .

[4]  R. Jamasbi,et al.  Growth inhibition and DNA damage induced by benzo[a]pyrene and formaldehyde in primary cultures of rat tracheal epithelial cells. , 1988, Mutation research.

[5]  M. Fujimura,et al.  Potentiation of allergic bronchoconstriction by repeated exposure to formaldehyde in guinea‐pigs in vivo , 2003, Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology.

[6]  R. Grafström.  In vitro studies of aldehyde effects related to human respiratory carcinogenesis. , 1990, Mutation research.

[7]  M. Casanova,et al.  Pharmacodynamics of formaldehyde: applications of a model for the arrest of DNA replication by DNA-protein cross-links. , 1999, Toxicology and applied pharmacology.

[8]  W. Ross,et al.  Relationship between DNA damage and survival in formaldehyde-treated mouse cells. , 1980, Mutation research.

[9]  W. Hennink,et al.  Identification of Formaldehyde-induced Modifications in Proteins , 2004, Journal of Biological Chemistry.

[10]  Eunha Oh,et al.  Effects of benzo(a)pyrene on protein expression in Jurkat T‐cells , 2004, Proteomics.

[11]  A. Borchert,et al.  Regulation of enzymatic lipid peroxidation: the interplay of peroxidizing and peroxide reducing enzymes. , 2002, Free radical biology & medicine.

[12]  Wei Wang,et al.  [Effects of formaldehyde on germ cells of male mice]. , 2003, Wei sheng yan jiu = Journal of hygiene research.

[13]  R. Desnick,et al.  Hydroxymethylbilane synthase: complete genomic sequence and amplifiable polymorphisms in the human gene. , 1993, Genomics.

[14]  J. Fagan,et al.  Quantitation of oxidative damage to tissue proteins. , 1999, The international journal of biochemistry & cell biology.

[15]  A. Konno,et al.  Effect of formaldehyde on the expression of adhesion molecules in nasal microvascular endothelial cells: the role of formaldehyde in the pathogenesis of sick building syndrome , 2002, Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology.

[16]  Roberto Colombo,et al.  Protein carbonyl groups as biomarkers of oxidative stress. , 2003, Clinica chimica acta; international journal of clinical chemistry.

[17]  R. Corvi.  Genomics: An In Vitro Toxicology Point of View , 2002, Alternatives to laboratory animals : ATLA.

[18]  E. Niki,et al.  Cytotoxic effect of formaldehyde with free radicals via increment of cellular reactive oxygen species. , 2005, Toxicology.

[19]  C. Bayerl,et al.  Low concentrations of formaldehyde induce DNA damage and delay DNA repair after UV irradiation in human skin cells , 2004, Experimental dermatology.

[20]  M. Taira,et al.  Hexachlorobenzene impairs glucose metabolism in a rat model of porphyria cutanea tarda: a mechanistic approach , 2004, Archives of Toxicology.

[21]  S. John,et al.  Mouse natriuretic peptide receptor 3 gene maps to proximal Chromosome 15 , 1997, Mammalian Genome.

[22]  G. Cosma,et al.  Benzo[a]pyrene- and formaldehyde-induced DNA damage and repair in rat tracheal epithelial cells. , 1988, Toxicology.

[23]  S. Hecht,et al.  Reactions of formaldehyde plus acetaldehyde with deoxyguanosine and DNA: formation of cyclic deoxyguanosine adducts and formaldehyde cross-links. , 2003, Chemical research in toxicology.

[24]  K. Doi,et al.  Rat strain difference in histology and expression of Th1- and Th2-related cytokines in nasal mucosa after short-term formaldehyde inhalation. , 2003, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[25]  C. Harris,et al.  Formaldehyde damage to DNA and inhibition of DNA repair in human bronchial cells. , 1983, Science.

[26]  H. Nakazawa,et al.  A case of sick building syndrome in a Japanese office worker. , 2005, Industrial health.

[27]  W. Zhong,et al.  Formaldehyde-induced DNA adducts as biomarkers of in vitro human nasal epithelial cell exposure to formaldehyde. , 2004, Mutation research.

[28]  R. Tice,et al.  A simple technique for quantitation of low levels of DNA damage in individual cells. , 1988, Experimental cell research.

[29]  Shawn D. Chase,et al.  Rapid, fluorimetric-liquid chromatographic determination of malondialdehyde in biological samples. , 2002, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[30]  M. Çiftçi,et al.  Effect of melatonin on enzyme activities of glutathione reductase from human erythrocytes in vitro and from rat erythrocytes in vivo. , 2006, European journal of pharmacology.

[31]  R. Dempsey,et al.  Phospholipase A2, hydroxyl radicals, and lipid peroxidation in transient cerebral ischemia. , 2003, Antioxidants & redox signaling.

[32]  C. Harris,et al.  Repair of DNA damage caused by formaldehyde in human cells. , 1984, Cancer research.