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Mechanical stiffness softening and cell adhesion are coordinately regulated by ERM dephosphorylation in KG-1 cells

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Abstract

Mechanical stiffness is closely related to cell adhesion and rounding in some cells. In leukocytes, dephosphorylation of ezrin/radixin/moesin (ERM) proteins is linked to cell adhesion events. To elucidate the relationship between surface stiffness, cell adhesion, and ERM dephosphorylation in leukocytes, we examined the relationship in the myelogenous leukemia line, KG-1, by treatment with modulation drugs. KG-1 cells have ring-shaped cortical actin with microvilli as the only F-actin cytoskeleton, and the actin structure constructs the mechanical stiffness of the cells. Phorbol 12-myristate 13-acetate and staurosporine, which induced cell adhesion to fibronectin surface and ERM dephosphorylation, caused a decrease in surface stiffness in KG-1 cells. Calyculin A, which inhibited ERM dephosphorylation and had no effect on cell adhesion, did not affect surface stiffness. To clarify whether decreasing cell surface stiffness and inducing cell adhesion are equivalent, we examined KG-1 cell adhesion by treatment with actin-attenuated cell softening reagents. Cytochalasin D clearly diminished cell adhesion, and high concentrations of Y27632 slightly induced cell adhesion. Only Y27632 slightly decreased ERM phosphorylation in KG-1 cells. Thus, decreasing cell surface stiffness and inducing cell adhesion are not equivalent, but these phenomena are coordinately regulated by ERM dephosphorylation in KG-1 cells.

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References

  1. Lecuit T, Lenne PF. Cell surface mechanics and the control of cell shape, tissue patterns and morphogenesis. Nat Rev Mol Cell Biol. 2007;8(8):633–44.

    Article  CAS  Google Scholar 

  2. Stewart MP, Helenius J, Toyoda Y, Ramanathan SP, Muller DJ, Hyman AA. Hydrostatic pressure and the actomyosin cortex drive mitotic cell rounding. Nature. 2011;469(7329):226–30.

    Article  CAS  Google Scholar 

  3. Kunda P, Pelling AE, Liu T, Baum B. Moesin controls cortical rigidity, cell rounding, and spindle morphogenesis during mitosis. Curr Biol. 2008;18(2):91–101.

    Article  CAS  Google Scholar 

  4. Shimizu Y, Haghparast SM, Kihara T, Miyake J. Cortical rigidity of round cells in mitotic phase and suspended state. Micron. 2012;43(12):1246–51.

    Article  CAS  Google Scholar 

  5. Eiraku M, Takata N, Ishibashi H, Kawada M, Sakakura E, Okuda S, et al. Self-organizing optic-cup morphogenesis in three-dimensional culture. Nature. 2011;472(7341):51–6.

    Article  CAS  Google Scholar 

  6. Okuda S, Takata N, Hasegawa Y, Kawada M, Inoue Y, Adachi T, et al. Strain-triggered mechanical feedback in self-organizing optic-cup morphogenesis. Sci Adv. 2018;4(11):eaau1354.

    Article  CAS  Google Scholar 

  7. Dai J, Sheetz MP. Mechanical properties of neuronal growth cone membranes studied by tether formation with laser optical tweezers. Biophys J. 1995;68(3):988–96.

    Article  CAS  Google Scholar 

  8. Wang N. Mechanical interactions among cytoskeletal filaments. Hypertension. 1998;32(1):162–5.

    Article  CAS  Google Scholar 

  9. Trickey WR, Vail TP, Guilak F. The role of the cytoskeleton in the viscoelastic properties of human articular chondrocytes. J Orthop Res. 2004;22(1):131–9.

    Article  Google Scholar 

  10. Sugitate T, Kihara T, Liu X-Y, Miyake J. Mechanical role of the nucleus in a cell in terms of elastic modulus. Curr Appl Phys. 2009;9(4, Supplement 1):e291–3.

    Article  Google Scholar 

  11. Haghparast SM, Kihara T, Miyake J. Distinct mechanical behavior of HEK293 cells in adherent and suspended states. PeerJ. 2015;3:e1131.

    Article  Google Scholar 

  12. Yamane J, Ohnishi H, Sasaki H, Narimatsu H, Ohgushi H, Tachibana K. Formation of microvilli and phosphorylation of ERM family proteins by CD43, a potent inhibitor for cell adhesion: cell detachment is a potential cue for ERM phosphorylation and organization of cell morphology. Cell Adh Migr. 2011;5(2):119–32.

    Article  Google Scholar 

  13. Tachibana K, Haghparast SM, Miyake J. Inhibition of cell adhesion by phosphorylated Ezrin/Radixin/Moesin. Cell Adh Migr. 2015;9(6):502–12.

    Article  CAS  Google Scholar 

  14. Tachibana K, Ohnishi H, Haghparast SMA, Kihara T, Miyake J. Activation of PKC induces leukocyte adhesion by the dephosphorylation of ERM. BIochem Biophys Res Commun. 2020;523(1):177–82.

    Article  CAS  Google Scholar 

  15. Brown MJ, Nijhara R, Hallam JA, Gignac M, Yamada KM, Erlandsen SL, et al. Chemokine stimulation of human peripheral blood T lymphocytes induces rapid dephosphorylation of ERM proteins, which facilitates loss of microvilli and polarization. Blood. 2003;102(12):3890–9.

    Article  CAS  Google Scholar 

  16. García-Ortiz A, Serrador JM. ERM proteins at the crossroad of leukocyte polarization, migration and intercellular adhesion. Int J Mol Sci. 2020;21(4):1502.

    Article  Google Scholar 

  17. Wojcikiewicz EP, Zhang X, Chen A, Moy VT. Contributions of molecular binding events and cellular compliance to the modulation of leukocyte adhesion. J Cell Sci. 2003;116(Pt 12):2531–9.

    Article  CAS  Google Scholar 

  18. Shimizu Y, Kihara T, Haghparast SM, Yuba S, Miyake J. Simple display system of mechanical properties of cells and their dispersion. PLoS ONE. 2012;7(3):e34305.

    Article  CAS  Google Scholar 

  19. Phan TKT, Shahbazzadeh F, Kihara T. Alpha-mangostin reduces mechanical stiffness of various cells. Hum Cell. 2020;33(2):347–55.

    Article  CAS  Google Scholar 

  20. Kato K, Umezawa K, Funeriu DP, Miyake M, Miyake J, Nagamune T. Immobilized culture of nonadherent cells on an oleyl poly(ethylene glycol) ether-modified surface. Biotechniques. 2003;35(5):1014–21.

    Article  CAS  Google Scholar 

  21. Phan TKT, Shahbazzadeh F, Pham TTH, Kihara T. Alpha-mangostin inhibits the migration and invasion of A549 lung cancer cells. PeerJ. 2018;6:e5027.

    Article  Google Scholar 

  22. Haghparast SM, Kihara T, Shimizu Y, Yuba S, Miyake J. Actin-based biomechanical features of suspended normal and cancer cells. J Biosci Bioeng. 2013;116(3):380–5.

    Article  CAS  Google Scholar 

  23. Jadhav S, Eggleton CD, Konstantopoulos K. A 3-D computational model predicts that cell deformation affects selectin-mediated leukocyte rolling. Biophys J. 2005;88(1):96–104.

    Article  CAS  Google Scholar 

  24. Rouven Brückner B, Pietuch A, Nehls S, Rother J, Janshoff A. Ezrin is a major regulator of membrane tension in epithelial cells. Sci Rep. 2015;5:14700.

    Article  Google Scholar 

  25. Staser K, Shew MA, Michels EG, Mwanthi MM, Yang FC, Clapp DW, et al. A Pak1-PP2A-ERM signaling axis mediates F-actin rearrangement and degranulation in mast cells. Exp Hematol. 2013;41(1):56-66.e2.

    Article  CAS  Google Scholar 

  26. Rullo J, Becker H, Hyduk SJ, Wong JC, Digby G, Arora PD, et al. Actin polymerization stabilizes α4β1 integrin anchors that mediate monocyte adhesion. J Cell Biol. 2012;197(1):115–29.

    Article  CAS  Google Scholar 

  27. Li Y, Harada T, Juang YT, Kyttaris VC, Wang Y, Zidanic M, et al. Phosphorylated ERM is responsible for increased T cell polarization, adhesion, and migration in patients with systemic lupus erythematosus. J Immunol. 2007;178(3):1938–47.

    Article  CAS  Google Scholar 

  28. Liu L, Schwartz BR, Lin N, Winn RK, Harlan JM. Requirement for RhoA kinase activation in leukocyte de-adhesion. J Immunol. 2002;169(5):2330–6.

    Article  CAS  Google Scholar 

  29. Silveira AAA, Dominical VM, Almeida CB, Chweih H, Ferreira WA Jr, Vicente CP, et al. TNF induces neutrophil adhesion via formin-dependent cytoskeletal reorganization and activation of β-integrin function. J Leukoc Biol. 2018;103(1):87–98.

    CAS  PubMed  Google Scholar 

  30. Uehata M, Ishizaki T, Satoh H, Ono T, Kawahara T, Morishita T, et al. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature. 1997;389(6654):990–4.

    Article  CAS  Google Scholar 

  31. Maekawa M, Ishizaki T, Boku S, Watanabe N, Fujita A, Iwamatsu A, et al. Signaling from Rho to the actin cytoskeleton through protein kinases ROCK and LIM-kinase. Science. 1999;285(5429):895–8.

    Article  CAS  Google Scholar 

  32. Narumiya S, Thumkeo D. Rho signaling research: history, current status and future directions. FEBS Lett. 2018;592(11):1763–76.

    Article  CAS  Google Scholar 

  33. Ishihara H, Ozaki H, Sato K, Hori M, Karaki H, Watabe S, et al. Calcium-independent activation of contractile apparatus in smooth muscle by calyculin-A. J Pharmacol Exp Ther. 1989;250(1):388–96.

    CAS  PubMed  Google Scholar 

  34. Saotome I, Curto M, McClatchey AI. Ezrin is essential for epithelial organization and villus morphogenesis in the developing intestine. Dev Cell. 2004;6(6):855–64.

    Article  CAS  Google Scholar 

  35. Pelaseyed T, Bretscher A. Regulation of actin-based apical structures on epithelial cells. J Cell Sci. 2018;131:20.

    Article  Google Scholar 

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Acknowledgements

We thank Ms. Thi Ly Do (The University of Kitakyushu) for her technical help in AFM analysis. The observation of F-actin structures of KG-1 cells was carried out using a Nikon C2 confocal laser scanning microscope at the Instrumentation Center, The University of Kitakyushu.

Funding

This work was supported by JSPS KAKENHI Grant Numbers JP23107006 and JP21K04797, and by a grant for Young Scientists, Institute of Environmental Science and Technology, The University of Kitakyushu.

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Authors and Affiliations

  1. Department of Life and Environment Engineering, Faculty of Environmental Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu, Kitakyushu, Fukuoka, 808-0135, Japan

    Takanori Kihara, Teru Matsumoto & Yoshihito Nakahashi

  2. Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan

    Kouichi Tachibana

  3. Department of Hematology and Oncology, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa, 259-1193, Japan

    Kouichi Tachibana

Authors
  1. Takanori Kihara
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  2. Teru Matsumoto
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  4. Kouichi Tachibana
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Contributions

Teru Matsumoto and Yoshihito Nakahashi perfomed AFM analysis. Kouichi Tachibana performed immunoblotting. Takanori Kihara performed cell adhesion experiments and is responsible for the whole of this study. All authors have read and agreed to the final version of the manuscript.

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Correspondence to Takanori Kihara.

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This work does not involve human participants or their data. In this study, we used KG-1 cell line RRID:CVCL_0374, which obtained from RIKEN BioResource Center (Ibaraki, Japan).

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Kihara, T., Matsumoto, T., Nakahashi, Y. et al. Mechanical stiffness softening and cell adhesion are coordinately regulated by ERM dephosphorylation in KG-1 cells. Human Cell 34, 1709–1716 (2021). https://doi.org/10.1007/s13577-021-00584-2

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  • DOI: https://doi.org/10.1007/s13577-021-00584-2

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