Metabolite Profiling to Monitor Organochlorine Pesticide Exposure in HepG2 Cell Culture
Gas chromatography coupled with time of flight mass spectrometry (GC/MS-TOF) was used to profile endogenous metabolites in HepG2 cell cultures to assess the metabolic changes induced by exposure to different organochlorine pesticides, their mixtures and controls (endosulfan, lindane, DDT and aldrin). Cells were cultured in DMEM with Glutamax at 37 °C with 5 % CO2 for 72 h and then exposed to each pesticide, pesticide mixture or DMSO (as a control) for 24 h, and finally, endogenous metabolites were extracted and analyzed using GC/MS-TOF. The experiment was repeated six times under the same cell passage and culture conditions. PCA, PLS-DA and ROC were performed to analyze the GC/MS-TOF data and identify potential biomarkers. Thirty-five explanatory metabolites were found in both PCA and PLS-DA models, where Q 2 was 0.86 and R 2 was 0.98. Univariate and multivariate ROC showed potential biomarkers for each treatment, suggesting a general toxic mechanism for organochlorine pesticides that is specific for each type of compound. These results confirmed the effect of OCPs in sugar and amino acid metabolism that are linked with the function of cytochrome P450 in reductive dechlorination and oxidative stress.
KeywordsOrganochlorine Pesticides Metabolite profiling Biomarkers Metabolomics
To Becas Generación de Bicentenario conv. 511 from Colciencias (Grant to Martha Zuluaga). To Programa de Jóvenes Investigadores e Innovadores from Colciencias and Universidad Tecnológica de Pereira (Grant to Melchor JJ and Tabares-Villa FA, contracts 5348/2013 and 5826/2013, respectively) and the West Coast Metabolomics Center for receiving Martha Zuluaga as a visitor scholar during 2014.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
- 3.Bonvallot N, Tremblay-Franco M, Chevrier C, Canlet C, Debrauwer L, Cravedi J-P, Cordier S (2014) Potential input from metabolomics for exploring and understanding the links between environment and health. J Toxicol Environ Health B Crit Rev 17(1):21–44. doi: 10.1080/10937404.2013.860318 CrossRefGoogle Scholar
- 8.Benarbia MEA, Macherel D, Faure S, Jacques C, Andriantsitohaina R, Malthièry Y (2013) Plasmatic concentration of organochlorine lindane acts as metabolic disruptors in HepG2 liver cell line by inducing mitochondrial disorder. Toxicol Appl Pharmacol 272(2):325–334. doi: 10.1016/j.taap.2013.06.006 CrossRefGoogle Scholar
- 15.Ruiz-aracama A, Peijnenburg A, Kleinjans J, Jennen D, Delft J Van, Hellfrisch C, Lommen A (2011) An untargeted multi-technique metabolomics approach to studying intracellular metabolites of HepG2 cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin. BMC Genom 12(251):1–19Google Scholar