Karmveer Yadav1, Syed Azmal Ali2, Ashok Kumar Mohanty2, Eshwarmoorthy Muthusamy3, Kesavan Subaharan4, Gautam Kaul5. 1. N.T. Lab-1, Division of Animal Biochemistry, ICAR-National Dairy Research Institute, Karnal, 132001, India. karmveery3@gmail.com. 2. Cell Biology and Proteomics Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal, 132001, Haryana, India. 3. Chemistry and Physics of Materials Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, 560064, India. 4. Division of Germplasm, Conservation and Utilisation, National Bureau of Agricultural Insect Resources, Bangalore, 560024, India. 5. N.T. Lab-1, Division of Animal Biochemistry, ICAR-National Dairy Research Institute, Karnal, 132001, India. gkndri@gmail.com.
Abstract
BACKGROUND: The 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. RESULTS: Through 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. CONCLUSIONS: Based 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.
BACKGROUND: The 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. RESULTS: Through 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. CONCLUSIONS: Based 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.
Entities:
Keywords:
Apoptosis; Cell morphology; Cytoskeleton dynamics; ER-mediated phagocytosis and nanoparticles
Authors: Jung Park; Sebastian Bauer; Karl Andreas Schlegel; Friedrich W Neukam; Klaus von der Mark; Patrik Schmuki Journal: Small Date: 2009-03 Impact factor: 13.281
Authors: Laura Larson; Serge Arnaudeau; Bruce Gibson; Wei Li; Ryoko Krause; Binghua Hao; James R Bamburg; Daniel P Lew; Nicolas Demaurex; Frederick Southwick Journal: Proc Natl Acad Sci U S A Date: 2005-01-25 Impact factor: 11.205
Authors: Cory Hanley; Janet Layne; Alex Punnoose; K M Reddy; Isaac Coombs; Andrew Coombs; Kevin Feris; Denise Wingett Journal: Nanotechnology Date: 2008-07-23 Impact factor: 3.874