Elasticity Mapping Analysis of Apical Cell Periphery Actin Structures of Normal Fibroblasts and Cervical Cancer Cells

Takanori Kihara, Takuji Yoshida, Seyed M. Ali Haghparast, Jun Miyake

Department of Mechanical Science and Bioengineering, Graduate School of Engineering Science, Osaka University, Osaka, Japan.

DOI: 10.4236/jasmi.2013.32015                

Abstract

The cell mechanical features are largely regulated by actin cytokeleton. By analyzing the mechanical features, it is possible to evaluate the characteristics of the complicated actin cytoskeleton in diverse cell types. In this study, we examined the sub-membrane mechanical structures of normal fibroblasts TIG-1 cells, and cervical cancer Hela cells using local elasticity mapping method of atomic force microscope. Especially we aimed at clarifying the regulatory mechanisms of sub-membrane actin structures in these cells by activation of actomyosin formation using calyculin A. This technique revealed that TIG-1 and Hela cells bore clearly different sub-membrane mechanical structures. TIG-1 cells had aligned stiff filamentous structures, whereas Hela cells had crooked and relatively soft filaments. The surface stiffness of TIG-1 cells increased slightly by actomyosin formation due to stiffness increase of the aligned filamentous structures. On the other hand, the surface stiffness of Hela cells increased by actomyosin formation due to upregulation of the apical actin filaments. Therefore, the structural and regulatory differences of the apical actin filaments could be demonstrated by atomic force microscopy elasticity mapping analysis.

Keywords

Atomic Force Microscopy; Actin Cytoskeleton; Elasticity Mapping Analysis; Mechanical Structure

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	J. M. Maloney, D. Nikova, F. Lautenschlager, E. Clarke, R. Langer, J. Guck and K. J. Van Vliet, “Mesenchymal Stem Cell Mechanics from the Attached to the Suspended State,” Biophysical Journal, Vol. 99, No. 8, 2010, pp. 2479-2487. doi:10.1016/j.bpj.2010.08.052
[2]	T. Kihara, S. M. Haghparast, Y. Shimizu, S. Yuba and J. Miyake, “Physical Properties of Mesenchymal Stem Cells Are Coordinated by the Perinuclear Actin Cap,” Biochemical and Biophysical Research Communications, Vol. 409, No. 1, 2011, pp. 1-6. doi:10.1016/j.bbrc.2011.04.022
[3]	S. Suresh, J. Spatz, J. P. Mills, A. Micoulet, M. Dao, C. T. Lim, M. Beil and T. Seufferlein, “Connections between Single-Cell Biomechanics and Human Disease States: Gastrointestinal Cancer and Malaria,” Acta Biomaterialia, Vol. 1, No. 1, 2005, pp. 15-30. doi:10.1016/j.actbio.2004.09.001
[4]	S. E. Cross, Y. S. Jin, J. Rao and J. K. Gimzewski, “Nanomechanical Analysis of Cells from Cancer Patients,” Nature Nanotechnology, Vol. 2, No. 12, 2007, pp. 780-783. doi:10.1038/nnano.2007.388
[5]	E. Evans and A. Yeung, “Apparent Viscosity and Cortical Tension of Blood Granulocytes Determined by Micropipet Aspiration,” Biophysical Journal, Vol. 56, No. 1, 1989, pp. 151-160. doi:10.1016/S0006-3495(89)82660-8
[6]	J. Guck, R. Ananthakrishnan, T. J. Moon, C. C. Cunningham and J. Kas, “Optical Deformability of Soft Biological Dielectrics,” Physical Review Letters, Vol. 84, No. 23, 2000, pp. 5451-5454. doi:10.1103/PhysRevLett.84.5451
[7]	M. Radmacher, M. Fritz, C. M. Kacher, J. P. Cleveland and P. K. Hansma, “Measuring the Viscoelastic Properties of Human Platelets with the Atomic Force Microscope,” Biophysical Journal, Vol. 70, No. 1, 1996, pp. 556-567. doi:10.1016/S0006-3495(96)79602-9
[8]	C. Rotsch, F. Braet, E. Wisse and M. Radmacher, “AFM Imaging and Elasticity Measurements on Living Rat Liver Macrophages,” Cell Biology International, Vol. 21, No. 11, 1997, pp. 685-696. doi:10.1006/cbir.1997.0213
[9]	H. Haga, S. Sasaki, K. Kawabata, E. Ito, T. Ushiki and T. Sambongi, “Elasticity Mapping of Living Fibroblasts by AFM and Immunofluorescence Observation of the Cytoskeleton,” Ultramicroscopy, Vol. 82, No. 1-4, 2000, pp. 253-258. doi:10.1016/S0304-3991(99)00157-6
[10]	R. Matzke, K. Jacobson and M. Radmacher, “Direct, High-Resolution Measurement of Furrow Stiffening during Division of Adherent Cells,” Nature Cell Biology, Vol. 3, No. 6, 2001, pp. 607-610. doi:10.1038/35078583
[11]	J. Dai and M. P. Sheetz, “Mechanical Properties of Neuronal Growth Cone Membranes Studied by Tether Formation with Laser Optical Tweezers,” Biophysical Journal, Vol. 68, No. 3, 1995, pp. 988-996. doi:10.1016/S0006-3495(95)80274-2
[12]	N. Wang, “Mechanical Interactions among Cytoskeletal Filaments,” Hypertension, Vol. 32, No. 1, 1998, pp. 162-165. doi:10.1161/01.HYP.32.1.162
[13]	W. R. Trickey, T. P. Vail and F. Guilak, “The Role of the Cytoskeleton in the Viscoelastic Properties of Human Articular Chondrocytes,” Journal of Orthopaedic Research, Vol. 22, No. 1, 2004, pp. 131-139. doi:10.1016/S0736-0266(03)0150-5
[14]	T. Sugitate, T. Kihara, X.-Y. Liu and J. Miyake, “Mechanical Role of the Nucleus in a Cell in Terms of Elastic Modulus,” Current Applied Physics, Vol. 9, No. 4, 2009, pp. e291-e293. doi:10.1016/j.cap.2009.06.020
[15]	L. Lu, S. J. Oswald, H. Ngu and F. C. Yin, “Mechanical Properties of Actin Stress Fibers in Living Cells,” Biophysical Journal, Vol. 95, No. 12, 2008, pp. 6060-6071. doi:10.1529/biophysj.108.133462
[16]	A. S. Maddox and K. Burridge, “RhoA Is Required for Cortical Retraction and Rigidity during Mitotic Cell Rounding,” The Journal of Cell Biology, Vol. 160, No. 2, 2003, pp. 255-265. doi:10.1083/jcb.200207130
[17]	P. Kunda, A. E. Pelling, T. Liu and B. Baum, “Moesin Controls Cortical Rigidity, Cell Rounding, and Spindle Morphogenesis during Mitosis,” Current Biology, Vol. 18, No. 2, 2008, pp. 91-101. doi:10.1016/j.cub.2007.12.051
[18]	Y. Shimizu, S. M. Haghparast, T. Kihara and J. Miyake, “Cortical Rigidity of Round Cells in Mitotic Phase and Suspended State,” Micron, Vol. 43, No. 12, 2012, pp. 1246-1251. doi:10.1016/j.micron.2012.03.011
[19]	M. Lekka, K. Pogoda, J. Gostek, O. Klymenko, S. Prauzner-Bechcicki, J. Wiltowska-Zuber, J. Jaczewska, J. Lekki and Z. Stachura, “Cancer Cell Recognition—Mechanical Phenotype,” Micron, Vol. 43, No. 12, 2012, pp. 1259-1266. doi:10.1016/j.micron.2012.01.019
[20]	S. M. A. Haghparast, T. Kihara, Y. Shimizu, S. Yuba and J. Miyake, “Actin-Based Biomechanical Features of Suspended Normal and Cancer Cells,” Journal of Bioscience and Bioengineering, 2013 (in press). doi:10.1016/j.jbiosc.2013.03.003
[21]	H. Ishihara, H. Ozaki, K. Sato, M. Hori, H. Karaki, S. Watabe, Y. Kato, N. Fusetani, K. Hashimoto and D. Uemura, “Calcium-Independent Activation of Contractile Apparatus in Smooth Muscle by Calyculin-A,” Journal of Pharmacology and Experimental Therapeutics, Vol. 250, No. 1, 1989, pp. 388-396.
[22]	M. Ohashi, S. Aizawa, H. Ooka, T. Ohsawa, K. Kaji, H. Kondo, T. Kobayashi, T. Noumura, M. Matsuo, Y. Mitsui, S. Murota, K. Yamamoto, H. Ito, H. Shimada and T. Utakoji, “A New Human Diploid Cell Strain, TIG-1, for the Research on Cellular Aging,” Experimental Gerontology, Vol. 15, No. 2, 1980, pp. 121-133. doi:10.1016/0531-5565(80)90083-2
[23]	H. Hertz, “über die Berührung Fester Elastischer Korper,” Journal für die Reine und Angewandte Mathematik, Vol. 92, No. 1881, pp. 156-171.
[24]	Y. Shimizu, T. Kihara, S. M. Haghparast, S. Yuba and J. Miyake, “Simple Display System of Mechanical Properties of Cells and Their Dispersion,” PLoS ONE, Vol. 7, No. 3, 2012, Article ID: e34305. doi:10.1371/journal.pone.0034305