Advertisement

Cytoskeletal Perturbing Drugs and Their Effect on Cell Elasticity

  • Martha E. Grady
  • Russell J. Composto
  • David M. EckmannEmail author
Conference paper
Part of the Conference Proceedings of the Society for Experimental Mechanics Series book series (CPSEMS)

Abstract

The cytoskeleton is primarily responsible for providing structural support, localization and transport of organelles, and intracellular trafficking. The structural support is supplied by actin filaments, microtubules, and intermediate filaments, which contribute to overall cell elasticity to varying degrees. We evaluate cell elasticity in five different cell types with drug-induced cytoskeletal derangements to probe how actin filaments and microtubules contribute to cell elasticity and whether it is conserved across cell type. Specifically, we measure elastic stiffness in chondrocytes, fibroblasts, endothelial cells, hepatocellular carcinoma, and fibrosarcoma using atomic force microscopy. We subject all five cell lines to two cytoskeletal destabilizers: cytochalasin D and nocodazole, which disrupt actin and microtubule polymerization, respectively. Non-cancer cells treated with cytochalasin D show a decrease of 60–80 % in moduli values compared to untreated cells of the same origin, whereas the nocodazole-treated cells show no change. Alternatively, cancer cells exhibit increased stiffness as well as stiffness variability when subjected to nocodazole. Overall, we demonstrate actin filaments contribute more to elastic stiffness than microtubules but this result is cell type dependent. Lastly, we show that disruption of microtubule dynamics affects cancer cell elasticity, suggesting therapeutic drugs targeting microtubules be monitored for significant elastic changes.

Keywords

Atomic force microscopy Cell mechanics Elasticity Cytoskeleton Cancer 

Notes

Acknowledgements

The authors gratefully acknowledge our funding sources: ONR Grant N000141612100 (DME), the Provost’s Postdoctoral Fellowship for Academic Diversity (MEG), NSF-NSEC Grant DMR08-32802 (RJC), and URF 4-000002-4820 (DME), which made this work possible. The work was performed at and supported by the Nano Bio Interface Center at the University of Pennsylvania through an instrumentation grant, DBI-0721913, and DMR-0425780. We also thank Judith Kandel for cell culture training and Dr. Matt Brukman and Dr. Matt Caporizzo for instrument support. We thank the following for their generous donations: Dr. Robert Mauck (chondrocytes), Dr. Ben Stanger (HUH-7), and Dr. Bruce Malkowicz (HT-1080).

References

  1. 1.
    Discher, D.E., Janmey, P., Wang, Y.L.: Tissue cells feel and respond to the stiffness of their substrate. Science 310(5751), 1139–1143 (2005)CrossRefGoogle Scholar
  2. 2.
    Bao, G., Suresh, S.: Cell and molecular mechanics of biological materials. Nat. Mater. 2(11), 715–725 (2003)CrossRefGoogle Scholar
  3. 3.
    Lee, G.Y.H., Lim, C.T.: Biomechanics approaches to studying human diseases. Trends Biotechnol. 25(3), 111–118 (2007)MathSciNetCrossRefGoogle Scholar
  4. 4.
    Rother, J., Noding, H., Mey, I., Janshoff, A.: Atomic force microscopy-based microrheology reveals significant differences in the viscoelastic response between malign and benign cell lines. Open Biol. 4(5), 140046 (2014)CrossRefGoogle Scholar
  5. 5.
    Nalam, P.C., Gosvami, N.N., Caporizzo, M.A., Composto, R.J., Carpick, R.W.: Nano-rheology of hydrogels using direct drive force modulation atomic force microscopy. Soft Matter 11(41), 8165–8178 (2015)CrossRefGoogle Scholar
  6. 6.
    Raman, A., Trigueros, S., Cartagena, A., Stevenson, A.P.Z., Susilo, M., Nauman, E., Contera, S.A.: Mapping nanomechanical properties of live cells using multi-harmonic atomic force microscopy. Nat. Nanotechnol. 6(12), 809–814 (2011)CrossRefGoogle Scholar
  7. 7.
    Caporizzo, M.A., Roco, C.M., Ferrer, M.C.C., Grady, M.E., Parrish, E., Eckmann, D.M., Composto, R.J.: Strain-rate dependence of elastic modulus reveals silver nanoparticle induced cytotoxicity. Nanobiomedicine 2 (2015). pii: 9Google Scholar
  8. 8.
    Ketene, A.N., Schmelz, E.M., Roberts, P.C., Agah, M.: The effects of cancer progression on the viscoelasticity of ovarian cell cytoskeleton structures. Nanomedicine 8(1), 93–102 (2012)Google Scholar
  9. 9.
    Corbin, E.A., Kong, F., Lim, C.T., King, W.P., Bashir, R.: Biophysical properties of human breast cancer cells measured using silicon MEMS resonators and atomic force microscopy. Lab Chip 15(3), 839–847 (2014)CrossRefGoogle Scholar
  10. 10.
    Moreno-Flores, S., Benitez, R., Vivanco, M.D., Toca-Herrera, J.L.: Stress relaxation and creep on living cells with the atomic force microscope: a means to calculate elastic moduli and viscosities of cell components. Nanotechnology 21(44), 445101 (2010)CrossRefGoogle Scholar
  11. 11.
    Kuznetsova, T.G., Starodubtseva, M.N., Yegorenkov, N.I., Chizhik, S.A., Zhdanov, R.I.: Atomic force microscopy probing of cell elasticity. Micron 38(8), 824–833 (2007)CrossRefGoogle Scholar
  12. 12.
    Rebelo, L.M., de Sousa, J.S., Mendes, J., Radmacher, M.: Comparison of the viscoelastic properties of cells from different kidney cancer phenotypes measured with atomic force microscopy. Nanotechnology 24(5), 055102 (2013)CrossRefGoogle Scholar
  13. 13.
    Park, S., Koch, D., Cardenas, R., Kas, J., Shih, C.K.: Cell motility and local viscoelasticity of fibroblasts. Biophys. J. 89(6), 4330–4342 (2005)CrossRefGoogle Scholar
  14. 14.
    Ribeiro, A.S., Khanna, P., Sukumar, A., Dong, C., Dahl, K.: Nuclear stiffening inhibits migration of invasive melanoma cells. Cell. Mol. Bioeng. 7(4), 544–551 (2014)CrossRefGoogle Scholar
  15. 15.
    Buda, A., Pignatelli, M.: Cytoskeletal network in colon cancer: from genes to clinical application. Int. J. Biochem. Cell Biol. 36(5), 759–765 (2004)CrossRefGoogle Scholar
  16. 16.
    Lindberg, U., Karlsson, R., Lassing, I., Schutt, C.E., Höglund, A.-S.: The microfilament system and malignancy. Semin. Cancer Biol. 18(1), 2–11 (2008)CrossRefGoogle Scholar
  17. 17.
    Grady, M.E., Composto, R.J., Eckmann, D.M.: Cell elasticity with altered cytoskeletal architectures across multiple cell types. J. Mech. Behav. Biomed. Mater. 61, 197–207 (2016)CrossRefGoogle Scholar
  18. 18.
    Trickey, W.R., Baaijens, F.P.T., Laursen, T.A., Alexopoulos, L.G., Guilak, F.: Determination of the Poisson's ratio of the cell: recovery properties of chondrocytes after release from complete micropipette aspiration. J. Biomech. 39(1), 78–87 (2006)CrossRefGoogle Scholar

Copyright information

© The Society for Experimental Mechanics, Inc. 2017

Authors and Affiliations

  • Martha E. Grady
    • 1
    • 2
  • Russell J. Composto
    • 1
  • David M. Eckmann
    • 2
    Email author
  1. 1.Department of Materials Science and EngineeringSchool of Engineering and Applied Science, University of PennsylvaniaPhiladelphiaUSA
  2. 2.Department of Anesthesiology and Critical CareSchool of Medicine, University of PennsylvaniaPhiladelphiaUSA

Personalised recommendations