Document Detail

Water quality assessment using the AREc32 reporter gene assay indicative of the oxidative stress response pathway.
MedLine Citation:
PMID:  23032559     Owner:  NLM     Status:  MEDLINE    
The reporter gene assay AREc32 is based on the induction of the Nrf2 mediated oxidative stress response pathway in the human breast cancer cell line MCF7, where eight copies of the antioxidant response element (ARE) are linked to a reporter gene encoding for luciferase. The Nrf2-ARE pathway is responsive to many chemicals that cause oxidative stress, among them a large number of pesticides and skin irritants. We adopted and validated the AREc32 bioassay for water quality testing. tert-Butylhydroquinone served as the positive control, phenol as the negative control and other reactive chemicals were assessed for their specificity. An environmentally relevant reference chemical, benzo(a)pyrene was the most potent inducer of all tested chemicals. The concentration causing an induction ratio (IR) of 1.5 (EC(IR1.5)) was chosen as the effect benchmark value. The assay was applied to 21 water samples ranging from sewage to drinking water, including secondary treatment and various tertiary treatment options (ozonation, biologically activated carbon filtration, membrane filtration, reverse osmosis, advanced oxidation, chlorination, chloramination). The samples were enriched by solid phase extraction. In most samples the oxidative stress response was far more sensitive than cytotoxicity. The primary and secondary treated effluent exceeded the effect threshold IR 1.5 at a relative enrichment factor (REF) of 1, i.e., the native samples were active. All tertiary treated samples were less potent and their EC(IR1.5) lay between REF 1 and 10. The Nrf2 pathway was induced at a REF of approximately 10 for surface waters and drinking water, and above this enrichment cytotoxicity took over in most samples and quenched the induction. The blank (ultrapure water run through the sample enrichment process) was cytotoxic at an REF of 100, which is the limit of concentrations range that can be evaluated. Treatment typically decreased both the cytotoxicity and oxidative stress response apart from drinking water treatment where chlorination caused an increase in oxidative stress response, presumably due to the formation of disinfection by-products. This study demonstrates the relevance and applicability of the oxidative stress response pathway for water quality monitoring.
Beate I Escher; Mriga Dutt; Erin Maylin; Janet Y M Tang; Simon Toze; C Roland Wolf; Matti Lang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of environmental monitoring : JEM     Volume:  14     ISSN:  1464-0333     ISO Abbreviation:  J Environ Monit     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-10-25     Completed Date:  2012-12-20     Revised Date:  2014-07-22    
Medline Journal Info:
Nlm Unique ID:  100968688     Medline TA:  J Environ Monit     Country:  England    
Other Details:
Languages:  eng     Pagination:  2877-85     Citation Subset:  IM    
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MeSH Terms
Antioxidant Response Elements / genetics
Biological Assay / methods*
Cell Line, Tumor
Drinking Water / chemistry
Environmental Monitoring / methods*
Genes, Reporter*
Luciferases / analysis,  metabolism
Oxidative Stress / genetics
Sewage / chemistry
Water Pollutants, Chemical / analysis*,  toxicity
Water Purification
Grant Support
10822//Cancer Research UK
Reg. No./Substance:
0/Drinking Water; 0/Sewage; 0/Water Pollutants, Chemical; EC 1.13.12.-/Luciferases

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