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Title (Primary) Hepatic co-cultures in vitro reveal suitable to detect Nrf2-mediated oxidative stress responses on the bladder carcinogen o-anisidine
Author Wewering, F.; Jouy, F.; Caliskan, S.; Kalkhof, S.; von Bergen, M.; Luch, A.; Zellmer, S.;
Journal Toxicology In Vitro
Year 2017
Department MOLSYB;
Volume 40
Language englisch;
POF III (all) F11;
Keywords o-Anisidine; ROS; Nuclear factor erythroid 2-related factor 2 (Nrf2); Co-culture; HepG2
UFZ wide themes RU3;
Abstract

The azo dye o-anisidine is known as an industrial and environmental pollutant. Metabolites of o-anisidine remain in the liver for > 24 h. However, the toxicological impact of o-anisidine on the liver and its individual cell types, e.g., hepatocytes and immune cells, is currently poorly understood.

A novel co-culture system, composed of HepG2 or Huh-7 cells, and differentiated THP-1 cells was used to study the metabolic capacity towards o-anisidine, and compared to primary murine hepatocytes which express high enzyme activities. As model compounds the carcinogenic arylamine o-anisidine and its non-carcinogenic isomer, p-anisidine, as well as caffeine were used.

Global proteome analysis revealed an activation of eIF2 and Nrf2-mediated oxidative stress response pathways only in co-cultures after treatment with o-anisidine. This was confirmed via detection of reactive oxygen species. In addition, the mitochondrial membrane potential decreased already after 3 h treatment of cells, which correlated with a decrease of ATP levels (R2 > 0.92). In the supernatant of co-cultured, but not single-cultured HepG2 and Huh-7 cells, o-anisidine caused increases of damage-associated proteins, such as HMGB1 (high mobility group box-1) protein.

In summary, only co-cultures of HepG2 and THP-1 cells predict o-anisidine induced stress responsive pathways, since the system has a higher sensitivity compared to single cultured cells.

ID 18664
Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=18664
Wewering, F., Jouy, F., Caliskan, S., Kalkhof, S., von Bergen, M., Luch, A., Zellmer, S. (2017):
Hepatic co-cultures in vitro reveal suitable to detect Nrf2-mediated oxidative stress responses on the bladder carcinogen o-anisidine
Toxicol. Vitro 40 , 153 - 160