Graphene Oxide Nanosheets Retard Cellular Migration via Disruption of Actin Cytoskeleton.

Graphene and graphene-based nanomaterials are broadly used for various biomedical applications due to their unique physiochemical properties. However, how graphene-based nanomaterials interact with biological systems has not been thoroughly studied. This study shows that graphene oxide (GO) nanosheets retard A549 lung carcinoma cell migration through nanosheet-mediated disruption of intracellular actin filaments. After GO nanosheets treatment, A549 cells display slower migration and the structure of the intracellular actin filaments is dramatically changed. It is found that GO nanosheets are capable of absorbing large amount of actin and changing the secondary structures of actin monomers. Large-scale all-atom molecular dynamics simulations further reveal the interactions between GO nanosheets and actin filaments at molecular details. GO nanosheets can insert into the interstrand gap of actin tetramer (helical repeating unit of actin filament) and cause the separation of the tetramer which eventually leads to the disruption of actin filaments. These findings offer a novel mechanism of GO nanosheet induced biophysical responses and provide more insights into their potential for biomedical applications.

[1]  Li Mu,et al.  Effects of Graphene Oxide and Oxidized Carbon Nanotubes on the Cellular Division, Microstructure, Uptake, Oxidative Stress, and Metabolic Profiles. , 2015, Environmental science & technology.

[2]  Carsten Kutzner,et al.  GROMACS 4:  Algorithms for Highly Efficient, Load-Balanced, and Scalable Molecular Simulation. , 2008, Journal of chemical theory and computation.

[3]  Yu Chen,et al.  Two-dimensional graphene analogues for biomedical applications. , 2015, Chemical Society reviews.

[4]  Gustavo K. Rohde,et al.  Altered Cell Mechanics from the Inside: Dispersed Single Wall Carbon Nanotubes Integrate with and Restructure Actin , 2012, Journal of functional biomaterials.

[5]  Jibin Song,et al.  Sequential Drug Release and Enhanced Photothermal and Photoacoustic Effect of Hybrid Reduced Graphene Oxide-Loaded Ultrasmall Gold Nanorod Vesicles for Cancer Therapy. , 2015, ACS nano.

[6]  Lin Zhao,et al.  Protein corona mitigates the cytotoxicity of graphene oxide by reducing its physical interaction with cell membrane. , 2015, Nanoscale.

[7]  W. L. Jorgensen,et al.  Comparison of simple potential functions for simulating liquid water , 1983 .

[8]  Lei Wang,et al.  Graphene oxide induces toll-like receptor 4 (TLR4)-dependent necrosis in macrophages. , 2013, ACS nano.

[9]  T. Wei,et al.  Inhibition of Cancer Cell Migration by Gold Nanorods: Molecular Mechanisms and Implications for Cancer Therapy , 2014 .

[10]  Hong-gang Zhao,et al.  Deguelin inhibits the migration and invasion of lung cancer A549 and H460 cells via regulating actin cytoskeleton rearrangement. , 2015, International journal of clinical and experimental pathology.

[11]  W. Kabsch,et al.  Atomic structure of the actin: DNase I complex , 1990, Nature.

[12]  G. Hummer,et al.  Water conduction through the hydrophobic channel of a carbon nanotube , 2001, Nature.

[13]  Yuichiro Maéda,et al.  The nature of the globular- to fibrous-actin transition , 2009, Nature.

[14]  Y. Wang,et al.  Actin reorganization as the molecular basis for the regulation of apoptosis in gastrointestinal epithelial cells , 2012, Cell Death and Differentiation.

[15]  Lay Poh Tan,et al.  Nanoparticles strengthen intracellular tension and retard cellular migration. , 2014, Nano letters.

[16]  Rongzheng Wan,et al.  Water-mediated signal multiplication with Y-shaped carbon nanotubes , 2009, Proceedings of the National Academy of Sciences.

[17]  M. Jordan,et al.  Microtubules and actin filaments: dynamic targets for cancer chemotherapy. , 1998, Current opinion in cell biology.

[18]  E. Egelman,et al.  A conformational change in the actin subunit can change the flexibility of the actin filament. , 1993, Journal of molecular biology.

[19]  M. Parrinello,et al.  Canonical sampling through velocity rescaling. , 2007, The Journal of chemical physics.

[20]  R. Zhou,et al.  Amino acid analogues bind to carbon nanotube via π-π interactions: comparison of molecular mechanical and quantum mechanical calculations. , 2012, The Journal of chemical physics.

[21]  D. Selkoe.  Alzheimer's disease: genes, proteins, and therapy. , 2001, Physiological reviews.

[22]  Daniel A. Fletcher,et al.  Cell mechanics and the cytoskeleton , 2010, Nature.

[23]  Brian D Holt,et al.  Carbon nanotubes reorganize actin structures in cells and ex vivo. , 2010, ACS nano.

[24]  R. Grosse,et al.  Pharmacological inhibition of actin assembly to target tumor cell motility. , 2014, Reviews of physiology, biochemistry and pharmacology.

[25]  Thomas D Pollard,et al.  Cellular Motility Driven by Assembly and Disassembly of Actin Filaments , 2003, Cell.

[26]  J. King,et al.  Aggregation of γ-crystallins associated with human cataracts via domain swapping at the C-terminal β-strands , 2011, Proceedings of the National Academy of Sciences.

[27]  Hanspeter Winkler,et al.  Molecular modeling of averaged rigor crossbridges from tomograms of insect flight muscle. , 2002, Journal of structural biology.

[28]  Ajay K. Royyuru,et al.  Free energy simulations reveal a double mutant avian H5N1 virus hemagglutinin with altered receptor binding specificity , 2009, J. Comput. Chem..

[29]  Masahide Takahashi,et al.  Cell biology of the movement of breast cancer cells: intracellular signalling and the actin cytoskeleton. , 2009, Cancer letters.

[30]  Huajian Gao,et al.  Graphene microsheets enter cells through spontaneous membrane penetration at edge asperities and corner sites , 2013, Proceedings of the National Academy of Sciences.

[31]  Haiping Fang,et al.  Destructive extraction of phospholipids from Escherichia coli membranes by graphene nanosheets. , 2013, Nature nanotechnology.

[32]  Ruhong Zhou,et al.  Adsorption of Villin Headpiece onto Graphene, Carbon Nanotube, and C60: Effect of Contacting Surface Curvatures on Binding Affinity , 2011 .

[33]  R. Zhou,et al.  Probing the self-assembly mechanism of diphenylalanine-based peptide nanovesicles and nanotubes. , 2012, ACS nano.

[34]  Alexander D. MacKerell,et al.  Extending the treatment of backbone energetics in protein force fields: Limitations of gas‐phase quantum mechanics in reproducing protein conformational distributions in molecular dynamics simulations , 2004, J. Comput. Chem..

[35]  Stefaan C De Smedt,et al.  High intracellular iron oxide nanoparticle concentrations affect cellular cytoskeleton and focal adhesion kinase-mediated signaling. , 2010, Small.

[36]  P. Kollman,et al.  Settle: An analytical version of the SHAKE and RATTLE algorithm for rigid water models , 1992 .

[37]  Xin Zhang,et al.  An efficient signal-on aptamer-based biosensor for adenosine triphosphate detection using graphene oxide both as an electrochemical and electrochemiluminescence signal indicator. , 2015, The Analyst.

[38]  Ruhong Zhou,et al.  Interactions between proteins and carbon-based nanoparticles: exploring the origin of nanotoxicity at the molecular level. , 2013, Small.

[39]  Gil Gonçalves,et al.  The effects of graphene oxide nanosheets localized on F-actin filaments on cell-cycle alterations. , 2013, Biomaterials.

[40]  R. Zhou,et al.  Urea-induced drying of carbon nanotubes suggests existence of a dry globule-like transient state during chemical denaturation of proteins. , 2010, The journal of physical chemistry. B.

[41]  H. Berendsen,et al.  Molecular dynamics with coupling to an external bath , 1984 .

[42]  Ki-Bum Lee,et al.  Design, synthesis, and characterization of graphene-nanoparticle hybrid materials for bioapplications. , 2015, Chemical reviews.

[43]  Ruhong Zhou,et al.  Observation of a dewetting transition in the collapse of the melittin tetramer , 2005, Nature.

[44]  Ruhong Zhou,et al.  Reduced Cytotoxicity of Graphene Nanosheets Mediated by Blood-Protein Coating. , 2015, ACS nano.

[45]  Berk Hess,et al.  LINCS: A linear constraint solver for molecular simulations , 1997 .

[46]  Ji-Xin Cheng,et al.  Highly sensitive transient absorption imaging of graphene and graphene oxide in living cells and circulating blood , 2015, Scientific Reports.

[47]  Ruhong Zhou,et al.  Hydrophobic Collapse in Multidomain Protein Folding , 2004, Science.

[48]  Ruhong Zhou,et al.  Thermal denaturing of mutant lysozyme with both the OPLSAA and the CHARMM force fields. , 2006, Journal of the American Chemical Society.

[49]  K Schulten,et al.  VMD: visual molecular dynamics. , 1996, Journal of molecular graphics.