Proteome-wide analysis reveals molecular pathways affected by AgNP in a ROS-dependent manner

Abstract

The use of mass spectrometry-based proteomics has been increasingly applied in nanomaterials risk assessments as it provides a proteome-wide overview of the molecular disturbances induced by its exposure. Here, we used this technique to gain detailed molecular insights into the role of ROS as an effector of AgNP toxicity, by incubating Bend3 cells with AgNP in the absence or presence of an antioxidant N-acetyl L-cystein (NAC). ROS generation is a key player in AgNP-induced toxicity, as cellular homeostasis was kept in the presence of NAC. By integrating MS/MS data with bioinformatics tools, in the absence of NAC, we were able to pinpoint precisely which biological pathways were affected by AgNP. Cells respond to AgNP-induced ROS generation by increasing their antioxidant pool, via NRF2 pathway activation. Additionally, cell proliferation-related pathways were strongly inhibited in a ROS-dependent manner. These findings reveal important aspects of the AgNP mechanism of action at the protein level.

Keywords:

• Silver nanoparticles
• oxidative stress
• N-Acetyl L-cysteine
• metal uptake
• proteomics

Acknowledgements

We also thank the VILLUM Center for Bioanalytical Sciences and PRO-MS: Danish National Mass Spectrometry Platform for Functional Proteomics (grant no. 5072-00007B) for support to the infrastructure.

Disclosure statement

The authors report no conflicts of interest.

Additional information

Funding

This study has received financial support from the European Research Council (ERC) under the European Union’s Horizon 2020 Research and Innovation Programme (Grant agreement No. 646603).