title: Quantification for total demethylation potential of environmental samples utilizing the EGFP reporter gene.
abstract: The demethylation potential of pollutants is arguably an innate component of their toxicity in environmental samples. A method was developed for determining the total demethylation potential of food samples (TDQ). The demethylation epigenetic toxicity was determined using the Hep G2 cell line transfected with pEGFP-C3 plasmids containing a methylated promoter of the EGFP reporter gene. The total demethylation potential of the sample extracts (the 5-AZA-CdR demethylation toxic equivalency) can be quantified within one week by using a standard curve of the 5-AZA-CdR demethylation agent. To explore the applicability of TDQ for environmental samples, 17 groundwater samples were collected from heavy polluted Kuihe river and the total demethylation potentials of the sample extracts were measured successfully. Meaningful demethylation toxic equivalencies ranging from 0.00050 to 0.01747μM were found in all groundwater sample extracts. Among 19 kinds of inorganic substance, As and Cd played important roles for individual contribution to the total demethylation epigenetic toxicity. The TDQ assay is reliable and fast for quantifying the DNA demethylation potential of environmental sample extracts, which may improve epigenetic toxicity evaluations for human risk assessment, and the consistent consuming of groundwater alongside the Kuihe river pose unexpected epigenetic health risk to the local residents.
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Qian
authors.affiliation: College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China.
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China.
authors.forename: Zhan-Lu
authors.initials: ZL
authors.lastname: Lv
authors.affiliation: Department of Chemistry, Umeå University, SE-901 87, Umeå, Sweden.
authors.forename: Mats
authors.initials: M
authors.lastname: Tysklind
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China; School of Public Health, Anhui Medical University, Hefei, Anhui 230032, China.
authors.forename: Bao
authors.initials: B
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China; School of Public Health, Anhui Medical University, Hefei, Anhui 230032, China.
authors.forename: Jia-Bing
authors.initials: JB
authors.lastname: Wu
authors.affiliation: School of Public Health, Anhui Medical University, Hefei, Anhui 230032, China.
authors.forename: Yong-Jian
authors.initials: YJ
authors.lastname: Yang
authors.affiliation: College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China.
authors.forename: Yi-Shu
authors.initials: YS
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China.
authors.forename: Fei-Fei
authors.initials: FF
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China.
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Duan
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China.
authors.forename: Yong-Jie
authors.initials: YJ
authors.lastname: Wei
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China.
authors.forename: Chun-Hui
authors.initials: CH
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China.
authors.forename: Li-Xin
authors.initials: LX
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China.
authors.forename: Jin-Liang
authors.initials: JL
authors.lastname: Zhang
authors.affiliation: Institute of Environmental Health and Related Product Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China.
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing 100012, China; Institute of Environmental Health and Related Product Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. Electronic address: xlwang@craes.org.cn.
authors.forename: Xian-Liang
authors.initials: XL
authors.lastname: Wang
mesh_terms: D001151:Arsenic / Q000032:analysis; D019175:DNA Methylation; D060766:Drinking Water / Q000032:analysis*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics*; D060587:Groundwater / Q000032:analysis*; D056945:Hep G2 Cells; D006801:Humans; D008670:Metals / Q000032:analysis; D014874:Water Pollutants, Chemical / Q000032:analysis*
chemical_list: D060766:Drinking Water; D008670:Metals; D014874:Water Pollutants, Chemical; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D001151:Arsenic
keywords: Demethylation potential; EGFP; Environmental sample; Epigenetic toxicity; Kuihe river