MSN, MWCNT and ZnO nanoparticle-induced CHO-K1 cell polarisation is linked to cytoskeleton ablation

Karmveer Yadav,Ashok Kumar Mohanty,Gautam Kaul,Syed Azmal Ali,Eshwarmoorthy Muthusamy,Kesavan Subaharan

doi:10.1186/s12951-021-00779-7

Abstract

BackgroundThe cellular response to nanoparticles (NPs) for the mechanical clue and biochemical changes are unexplored. Here, we provide the comprehensive analysis of the Chinese Hamster Ovary (CHO-K1) cell line to study cell behaviour following the exposure of mesoporous silica nanoparticle (MSN), multiwall carbon nanotubes (MWCNTs), and zinc oxide (ZnO) NPs.ResultsThrough the high-throughput proteomic study, we observed that the effect of NPs is alone not restricted to cell viability but also on cell polarisation. In the case of MSN, no drastic changes were observed in cellular morphology, but it upregulated chaperons that might prevent protein aggregation. However, MWCNT showed elongated cell appearance with numerous cytoplasmic vacuoles, and induce lamellipodia formation through actin polymerisation. The cytoskeleton remodelling was accompanied by the increased expression of Dlc-1, cofilin and Rac1 proteins. While ZnO NPs resulted in the rounded cell morphology along with nuclear abnormalities. The proteome analysis revealed that UBXN11 control cell roundness and DOCK3 leads to actin stress fibre formation and finally, loss of cell adhesion. It enhances the expression of catastrophic DNA damage and apoptotic proteins, which was unrecoverable even after 72 h, as confirmed by the colony formation assay. All three NPs trigger over-expression of the endocytic pathway, ubiquitination, and proteasomal complex proteins. The data indicate that ZnO and MSN entered into the cells through clathrin-mediated pathways; whereas, MWCNT invades through ER-mediated phagocytosis.ConclusionsBased on the incubation and concentration of NPs, our work provides evidence for the activation of Rac-Rho signalling pathway to alter cytoskeleton dynamics. Our results assist as a sensitive early molecular readout for nanosafety assessment.

Highlights

Nanotechnology has emerged as a multidimensional discipline getting utilised extensively in everyday life and biomedical field
The results prove that zinc oxide (ZnO) NPs have higher toxicity than multiwall carbon nanotubes (MWCNTs) and mesoporous silica nanoparticle (MSN). 5 μg/mL dose of ZnO NPs selected as High Observed Adverse Effect Level (HOAEL) because, at this concentration, we found a 70% decrease in cell viability
The results showed a significant decrease (1664 ± 451.7 μm2) in surface area on exposure with ZnO5 4259 ± 81.23 μm2 (P < 0.01) whereas, the surface area was significantly increased after exposure to MWCNT5 6491 ± 355.6 μm2 (P < 0.05)

Summary

Results

Through the high-throughput proteomic study, we observed that the effect of NPs is alone not restricted to cell viability and on cell polarisation. While ZnO NPs resulted in the rounded cell morphology along with nuclear abnormalities. The proteome analysis revealed that UBXN11 control cell roundness and DOCK3 leads to actin stress fibre formation and loss of cell adhesion. It enhances the expression of catastrophic DNA damage and apoptotic proteins, which was unrecoverable even after 72 h, as confirmed by the colony formation assay. The data indicate that ZnO and MSN entered into the cells through clathrin-mediated pathways; whereas, MWCNT invades through ER-mediated phagocytosis

Conclusions

Introduction

Materials and methods

Results and discussion

Conclusion