Cadmium exposure down-regulates 8-oxoguanine DNA glycosylase expression in rat lung and alveolar epithelial cells.
The current study tested the hypothesis that the pulmonary carcinogenic potential of cadmium (Cd) is related to its ability to inhibit the expression (mRNA and protein) and activity of 8-oxoguanine-DNA glycosylase (OGG1), a base excision repair (BER) enzyme that functions to preferentially excise pre-mutagenic 7,8-dihydro-8-oxoguanine (8-oxoG) from DNA. We demonstrate that a single Cd aerosol exposure of adult male Lewis rats causes time- and dose-dependent down-regulation in the pulmonary levels of rOGG1 mRNA and OGG1 protein, quantified by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assays and western analyses, respectively. Immunohistochemical studies confirmed that Cd inhalation reduces the relative amount of OGG1 in lungs of exposed animals without altering its over-all distribution within the lung, which appears to be more prominent within the alveolar epithelium. In agreement with our in vivo studies, we show that OGG1 expression is also attenuated in alveolar epithelial cell cultures exposed to CdCl(2) either acutely or by repeated passaging in Cd-containing medium. The effects caused by Cd were observed in cells that show no loss in viability, as assessed by colony forming ability, the MTT assay, and propidium iodide membrane permeability studies. Nuclear extracts prepared from Cd-treated cells also exhibit a reduction in the ability to nick a synthetic oligonucleotide containing 8-oxoG. We conclude from these studies that Cd causes suppression of OGG1 in the lung and that this mechanism may, in part, play a role in the Cd carcinogenic process.
Potts
Ryan J
RJ
Department of Biochemistry, University of Vermont College of Medicine, Burlington, VT 05405-0068, USA.
Watkin
Richard D
RD
Hart
Beth A
BA
D000818:Animals; D015153:Blotting, Western; D002104:Cadmium / Q000633:toxicity*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D043382:DNA-Formamidopyrimidine Glycosylase; D015536:Down-Regulation / Q000187:drug effects*; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D007150:Immunohistochemistry; D008168:Lung / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D009699:N-Glycosyl Hydrolases / Q000235:genetics* / Q000378:metabolism*; D011419:Propidium / Q000494:pharmacology; D011650:Pulmonary Alveoli / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D011917:Rats, Inbred Lew; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013778:Tetrazolium Salts; D013844:Thiazoles; D014410:Tumor Stem Cell Assay
D013778:Tetrazolium Salts; D013844:Thiazoles; D002104:Cadmium; D011419:Propidium; D009699:N-Glycosyl Hydrolases; D043382:DNA-Formamidopyrimidine Glycosylase; C022616:thiazolyl blue