title: DNA methylation analysis in rat kidney epithelial cells exposed to 3-MCPD and glycidol.
abstract: 3-Monochloropropane-1,2-diol (3-MCPD) is a well-known food processing contaminant that has been regarded as a rat carcinogen, which is known to induce Leydig-cell and mammary gland tumors in males, as well as kidney tumors in both genders. 3-MCPD is highly suspected to be a non-genotoxic carcinogen. 2,3-Epoxy-1-propanol (glycidol) can be formed via dehalogenation from 3-MCPD. We aimed to investigate the cytotoxic effects of 3-MCPD and glycidol, then to demonstrate the possible epigenetic mechanisms with global and gene-specific DNA methylation in rat kidney epithelial cells (NRK-52E). IC50value of 3-MCPD was determined as 48 mM and 41.39 mM, whereas IC50value of glycidol was 1.67 mM and 1.13 mM by MTT and NRU test, respectively. Decreased global DNA methylation at the concentrations of 100 μM and 1000 μM for 3-MCPD and 100 μM and 500 μM for glycidol were observed after 48 h exposure by using 5-methylcytosine (5-mC) ELISA kit. Methylation changes were detected in promoter regions of c-myc and Rassf1a in 3-MCPD and glycidol treated NRK-52E cells by using methylation-specific PCR (MSP), whereas changes on gene expression of c-myc and Rassf1a were observed by using real-time PCR. However, e-cadherin, p16, VHL and p15 genes were unmethylated in their CpG promoter regions in response to treatment with 3-MCPD and glycidol. Alterations in DNA methylation might be key events in the toxicity of 3-MCPD and glycidol.
authors.affiliation: a Department of Pharmaceutical Toxicology , Faculty of Pharmacy, Istanbul University , Istanbul , Turkey.
authors.forename: Mine
authors.initials: M
authors.lastname: Senyildiz
authors.affiliation: a Department of Pharmaceutical Toxicology , Faculty of Pharmacy, Istanbul University , Istanbul , Turkey.
authors.forename: Buket
authors.initials: B
authors.lastname: Alpertunga
authors.affiliation: a Department of Pharmaceutical Toxicology , Faculty of Pharmacy, Istanbul University , Istanbul , Turkey.
authors.forename: Sibel
authors.initials: S
authors.lastname: Ozden
mesh_terms: D000818:Animals; D002273:Carcinogens / Q000633:toxicity*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002629:Chemosterilants / Q000633:toxicity*; D018899:CpG Islands / Q000187:drug effects; D019175:DNA Methylation / Q000187:drug effects*; D044127:Epigenesis, Genetic / Q000187:drug effects*; D004852:Epoxy Compounds / Q000633:toxicity*; D005786:Gene Expression Regulation / Q000187:drug effects; D016259:Genes, myc / Q000187:drug effects; D020128:Inhibitory Concentration 50; D007684:Kidney Tubules / Q000187:drug effects* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020005:Propanols / Q000633:toxicity*; D051381:Rats; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism; D000517:alpha-Chlorohydrin / Q000633:toxicity*
chemical_list: D002273:Carcinogens; D002629:Chemosterilants; D004852:Epoxy Compounds; D020005:Propanols; C560857:RASSF1 protein, rat; D025521:Tumor Suppressor Proteins; D000517:alpha-Chlorohydrin; C004312:glycidol
keywords: 3-MCPD; DNA methylation; NRK-52 cells; cytotoxicity; glycidol