Development of normal human colon cell cultures to identify priority unregulated disinfection by-products with a carcinogenic potential.
Research was initiated to develop an in vitro system to identify disinfection by-products with a potential to transform normal human colonocytes into malignant cells. Tribromomethane and bromochloroacetic acid, rodent colon carcinogens, dibromonitromethane and tribromonitromethane, recently identified in drinking water, and azoxymethane, a classic colon carcinogen, were tested for the ability to transform NCM460 cells. The chronic toxicity was determined for the series of trihalomethanes, haloacetic acids and halonitromethanes as well as NCM460 cell enzymatic capabilities. The order of cytotoxicity was halonitromethanes > haloacetic acids > trihalomethanes. Cytotoxicity within a series increased with the degree of bromination and decreased with the molecular weight. The genotoxicity profile was similar to that for cytotoxicity. Enzymatic analysis demonstrated that NCM460 cells possess glutathione-S transerase-1-1 and CYP450 activity similar to that measured in the large intestine. NCM460 cells were exposed to 10(-6) M of the test chemicals for three days. While NCM460 cells from all treatments had the ability to grow in soft agar to some extent, only cells exposed to azoxymethane or tribromomethane were able to grow in media lacking serum and growth factors. When sub cultured, NCM460 cells exposed to 10(-9) M azoxymethane for three weeks formed colonies with morphology distinct from untreated cells.
DeAngelo
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National Health and Environmental Effects Research Laboratory, United States Environmental Protection Agency, 109 T.W. Alexander Drive, Research Triangle Park, NC 27711, USA. deangelo.anthony@epa.gov
Jones
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Moyer
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D000085:Acetates / Q000494:pharmacology; D001397:Azoxymethane / Q000494:pharmacology; D002273:Carcinogens / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D002471:Cell Transformation, Neoplastic / Q000187:drug effects*; D002478:Cells, Cultured; D003106:Colon / Q000166:cytology* / Q000201:enzymology; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D006842:Hydrocarbons, Brominated / Q000494:pharmacology; D020128:Inhibitory Concentration 50; D013997:Time Factors; D022882:Trihalomethanes / Q000494:pharmacology
D000085:Acetates; D002273:Carcinogens; D006842:Hydrocarbons, Brominated; D022882:Trihalomethanes; D003577:Cytochrome P-450 Enzyme System; C099813:bromochloroacetic acid; D005982:Glutathione Transferase; D001397:Azoxymethane; C015044:bromoform