title: Evaluation of 8-hydroxy-2'-deoxyguanosine (8-OHdG) adduct levels and DNA strand breaks in human peripheral blood lymphocytes exposed in vitro to polycyclic aromatic hydrocarbons with or without animal metabolic activation.
abstract: The polycyclic aromatic hydrocarbons (PAHs) dibenzo(a,h)anthracene, benzo(ghi)perylene, benzo(b)fluoranthene and benzo(a)pyrene have been identified in urban air from Mexico City and some of them are classified as human carcinogens. In the present study, human peripheral blood lymphocytes were exposed in vitro to different concentrations of PAHs with (+S9) or without (-S9) metabolic activation. The genotoxic and cytotoxic effects of each PAH were examined with an alkaline comet assay and trypan blue dye exclusion, and oxidative DNA damage was determined via the detection of 8-hydroxy-2'-deoxyguanosine (8-OhdG) adduct levels by enzyme-linked immunosorbent assay (ELISA). The DNA damage was evaluated with two genotoxicity parameters: the frequency of comets and the comet tail length. Concentrations of 20, 40, 80, 160 and 320 µM DB(a,h)A-S9; 20, 40, 80, 160 and 240 µM B(ghi)P-S9; 20, 30, 40, 60 and 80 µM B(b)F-S9; and 80 µM B(a)P-S9 for 24 h induced a small but significant increase in the means of comet frequency, in the tail length and in the 8-oHDg levels in relation to the control (0.5% DMSO-S9). However, all PAHs+S9 produced a more significant increase in DNA strand breaks and the level of 8-OHdG compared with the control (0.5% DMSO+S9), with a concentration-effect relationship. The viability of lymphocytes exposed to all PAHs-S9 and PAHs+S9 was not modified compared with the control. The results of this study demonstrate that the comet and ELISA are rapid, suitable and sensitive methods to detect in vitro PAH-induced DNA damage in human peripheral lymphocytes.
authors: Laboratorio de Citogenética Ambiental, y Química Atmosférica Centro de Ciencias de la Atmósfera, Universidad Nacional Autónoma de México, Ciudad Universitaria Coyoacán, México.
authors: Rodríguez-Romero María
authors: RR
authors: Isabel
authors: Gómez-Arroyo
authors: GA
authors: Sandra
authors: Villalobos-Pietrini
authors: VP
authors: Rafael
authors: Martínez-Valenzuela
authors: MV
authors: Carmen
authors: Cortés-Eslava
authors: CE
authors: Josefina
authors: Calderón-Ezquerro María
authors: CE
authors: del Carmen
authors: García-Martínez
authors: GM
authors: Rocío
authors: Arenas-Huertero
authors: AH
authors: Francisco
authors: Calderón-Segura María
authors: CS
authors: Elena
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000393:Air Pollutants / Q000378:metabolism / Q000633:toxicity*; D000818:Animals; D001711:Biotransformation; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D020552:Comet Assay; D018736:DNA Adducts / Q000378:metabolism*; D053960:DNA Breaks; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008099:Liver / Q000201:enzymology; D008214:Lymphocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D018384:Oxidative Stress / Q000187:drug effects; D011084:Polycyclic Aromatic Hydrocarbons / Q000378:metabolism / Q000633:toxicity*; D051381:Rats; D018570:Risk Assessment; D001335:Vehicle Emissions / Q000633:toxicity*
chemical_list: D000393:Air Pollutants; D018736:DNA Adducts; D011084:Polycyclic Aromatic Hydrocarbons; D001335:Vehicle Emissions; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine