Genotoxicity evaluation of multi-component mixtures of polyaromatic hydrocarbons (PAHs), arsenic, cadmium, and lead using flow cytometry based micronucleus test in HepG2 cells

Abstract

Some polyaromatic hydrocarbons (PAHs) and metals are known human carcinogens and the combined toxicity data of these co-contaminants are important for assessing their health risk. In this study, we have evaluated the combined genotoxicity, AhR activity and cell cycle parameters of four PAHs (benzo[a]pyrene (Ba]P), naphthalene (Nap), phenanthrene (Phe) and pyrene (Pyr)) and three metals (arsenic (As), cadmium (Cd), and lead (Pb)) in HepG2 cells using a flow cytometry based micronucleus (MN) test CAFLUX assay and nuclear fluorescence assay, respectively. The mixtures of B[a]P and metals induced a maximum of four fold increase in the MN formation compared to B[a]P alone. The higher combination of PAHs and metals did not significantly increase the MN formation. The mixtures of metals or non-carcinogenic PAHs were found to increase or decrease the aryl hydrocarbon receptor (AhR) activation of B[a]P in HepG2 cell based CAFLUX assay. Overall, the results showed that combined genotoxicity of PAHs and metals in HepG2 cells vary depending on the concentrations and number of the chemicals that are present in the mixtures and the effects of higher order combinations appear to be largely unpredictable from binary combinations. In this study, we have demonstrated the use of flow cytometry based MN test to screen the genotoxicity of environmental chemicals and its mixtures.