A proteomic approach to investigate AuNPs effects in Balb/3T3 cells.
Although gold nanoparticles (AuNPs) are currently used in several industrial products and biomedical applications, information about their biological effects is very limited. Thus, it is becoming crucial to assess their safety and adequately investigate the complexity of cell-nanoparticles interactions. In this work, the Balb/3T3 mouse fibroblast cell line was selected as an in vitro model to study the effects of AuNPs. Alteration of cellular processes and biochemical pathways caused by AuNPs exposure was investigated by analysing the differentially expressed proteome. Of interest was the difference observed in the protein pattern expression of cells exposed to AuNPs. It was found that 88 and 83 proteins were de-regulated after exposure to 5 and 15nm AuNPs, respectively. Analysis of the proteome revealed that AuNPs triggers several pathways related to cellular growth and proliferation, cell morphology, cell cycle regulation, cellular function and maintenance, oxidative stress, and inflammatory response. Moreover, SPR analysis showed an increase of ECM proteins biosynthesis in cells exposed to AuNPs. We observed by TEM analysis that NPs are internalized and confined mainly in autophagosomes. Endoplasmic reticulum stressed and modification at mitochondrial level occurred. This study aims to improve existing knowledge necessary for a correct assessment of the balance between AuNPs potential adverse and beneficial effects and might have important implications for biomedical applications (e.g. nanomedicine). To conclude proteomics link to system biology analysis is a valuable tool to understand and predict nanoparticles' toxicity, furthermore it has the potential to reveal pathways that may not be immediately evident with classical toxicological assays.
European Commission, Joint Research Centre, Institute for Health and Consumer Protection, Via Enrico Fermi 2749, I-21027 Ispra, Italy. Electronic address: sabrina.gioria@ec.europa.eu.
Sabrina
S
Gioria
European Commission, Joint Research Centre, Institute for Health and Consumer Protection, Via Enrico Fermi 2749, I-21027 Ispra, Italy.
Hubert
H
Chassaigne
INGM Istituto Nazionale di Genetica Molecolare, Via Francesco Sforza 28, I-20122 Milan, Italy.
Donatella
D
Carpi
European Commission, Joint Research Centre, Institute for Health and Consumer Protection, Via Enrico Fermi 2749, I-21027 Ispra, Italy.
Antonietta
A
Parracino
Italian National Institute of Health, Viale Regina Margherita 299, I-00161 Rome, Italy.
Stefania
S
Meschini
IRCCS Azienda Ospedaliera Universitaria San Martino, IST Istituto Nazionale per la Ricerca sul Cancro, Largo Rosanna Benzi, 10, I-16132 Genova, Italy.
Paola
P
Barboro
European Commission, Joint Research Centre, Institute for Health and Consumer Protection, Via Enrico Fermi 2749, I-21027 Ispra, Italy.
François
F
Rossi
D000818:Animals; D041702:BALB 3T3 Cells; D002458:Cell Fractionation; D002851:Chromatography, High Pressure Liquid; D024042:Collagen Type I / Q000096:biosynthesis / Q000235:genetics; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D005353:Fibronectins / Q000096:biosynthesis / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006046:Gold / Q000633:toxicity*; D006868:Hydrolysis; D007091:Image Processing, Computer-Assisted; D013058:Mass Spectrometry; D053768:Metal Nanoparticles / Q000633:toxicity*; D051379:Mice; D046529:Microscopy, Electron, Transmission; D010455:Peptides / Q000737:chemistry; D010766:Phosphorylation; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction / Q000187:drug effects; D020349:Surface Plasmon Resonance
D024042:Collagen Type I; D005353:Fibronectins; D010455:Peptides; D006046:Gold
Extracellular matrix (ECM); Gold nanoparticles (AuNPs); Liquid chromatography coupled to high-resolution mass spectrometry (LC–MS/MS); Surface plasma resonance (SPR); Transmission electron microscopy (TEM); Two-dimensional gel electrophoresis (2D-PAGE)