Titanium dioxide nanoparticles (TiO2 NPs) are widely found in consumer and industrial products, contributing to their prevalent presence in our surroundings. In this study, several miRNAs in the immuno-related pathways were found to be dysregulated in RAW264.7 cells after 24-h exposure to TiO2 NPs, including miR-29b-3p, which had not been previously found to be associated with the dysregulation of immunity after exposure to TiO2 NPs. The KEGG pathway and GO enrichment analysis suggested that miR-29b-3p functioned both in the T and B cell receptor signaling pathways. The NFAT5 gene was predicted to regulate miR-29b-3p using the MiRDB online database. The expression of miR-29b-3p and NFAT5 was found to be inversely correlated using qRT-PCR and western blotting analysis. Dual-luciferase reporter gene assays demonstrated the precise regulatory relationship between miR-29b-3p and NFAT5. The upregulation of miR-29b-3p was found to reinforce the apoptosis of cells, while no changes were found in terms of the cell cycle or cell proliferation, using MTT, cell apoptosis, and cycle detection experiments. Our results demonstrate that miR-29b-3p is involved in the response of RAW264.7 cells to exposure to TiO2, proving evidence for the further study of the toxicity and mechanisms of nano-TiO2 exposure.
Keywords: Bioinformatics; Cell apoptosis; Immuno-related pathways; NFAT5; RAW 264.7 macrophages; TiO2 nanoparticles; miR-29b-3p.