Cyclic stretch-induced mechanical stress to the cell nucleus inhibits ultraviolet radiation-induced DNA damage

  • Kazuaki NagayamaEmail author
  • Tomohiro Fukuei
Original Paper


Ultraviolet (UV) radiation exerts adverse effects on genome stability, alters the normal state of life, and causes several diseases by inducing DNA damage. Although mechanical stimulation such as stretching has significant effects on the prevention and treatment of diseases, its influence on nuclear morphology and/or intranuclear functions involving resistance to DNA damage remains unknown. Here, we investigated the effects of mechanical stimulation by cyclic stretching on nuclear morphology and resistance of DNA to UV damage in NIH3T3 fibroblasts. Adherent cells on silicone elastic membranes were subjected to ~ 10% cyclic uniaxial stretch at a frequency of 0.5 Hz for 12 h. As a result, the intracellular actin cytoskeleton and nucleus were found to be elongated and aligned in the direction of zero normal strain (~ 62° with respect to the stretch direction) in an actomyosin tension-dependent manner. The nuclei of the stretched cells were dramatically compressed by the reorganized actin stress fibers located on their apical and both sides, and a significant increase in the intranuclear DNA density was observed. Intercellular tension, as assessed with live cell atomic force microscopy imaging, also increased following exposure to cyclic stretch. The UV radiation-induced DNA damage, estimated from the fluorescence intensity of the phospho-histone γ-H2AX, significantly decreased in these stretched cells. These results indicate that the cyclic stretch-induced morphological changes in the nucleus may improve the UV radiation resistance of cells, probably owing to the intracellular force-induced condensation of chromatin. To our knowledge, this is the first study to demonstrate the inhibition of the UV radiation-induced DNA damage by mechanical stimulation.


Cell biomechanics Mechanobiology Nuclear mechanotransduction DNA damage 



This work was supported in part by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology, Japan (Nos. 17H02077 and 19K22944); the Naito foundation, Japan; the Takahashi industrial and economic research foundation, Japan; and AMED-CREST from Japan Agency for Medical Research and Development, AMED. The authors would like to thank Mrs. Akiko Sato for her technical help in the image analysis.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest with regard to this manuscript.

Supplementary material

10237_2019_1224_MOESM1_ESM.pdf (2.3 mb)
Supplementary material 1 (PDF 2332 kb)


  1. Alapetite C, Wachter T, Sage E, Moustachi E (1996) The use of the comet assay to detect DNA-repair deficiencies in human fibroblasts exposed to UVC, UVB, UVA and gamma-rays. Int J Radiat Biol 69:359–369CrossRefGoogle Scholar
  2. Boutahar N, Guignandon A, Vico L, Lafage-Proust MH (2004) Mechanical strain on osteoblasts activates autophosphorylation of focal adhesion kinase and proline-rich tyrosine kinase 2 tyrosine sites involved in ERK activation. J Biol Chem 279(29):30588–30599CrossRefGoogle Scholar
  3. Burton K, Park JH, Taylor DL (1999) Keratocytes generate traction forces in two phases. Mol Biol Cell 10(11):3745–3769CrossRefGoogle Scholar
  4. Dartsch PC, Hammerle H, Betz E (1986) Orientation of cultured arterial smooth muscle cells growing on cyclically stretched substrates. Acta Anat (Basel) 125:108–113CrossRefGoogle Scholar
  5. Galbraith CG, Sheetz MP (1997) A micromachined device provides a new bend on fibroblast traction forces. Proc Natl Acad Sci USA 94(17):9114–9118CrossRefGoogle Scholar
  6. Gaston J, Quinchia Rios B, Bartlett R, Berchtold C, Thibeault SL (2012) The response of vocal fold fibroblasts and mesenchymal stromal cells to vibration. PLoS ONE 7(2):e30965CrossRefGoogle Scholar
  7. Ghazanfari S, Tafazzoli-Shadpour M, Shokrgozar MA (2009) Effects of cyclic stretch on proliferation of mesenchymal stem cells and their differentiation to smooth muscle cells. Biochem Biophys Res Commun 388(3):601–605CrossRefGoogle Scholar
  8. Gorgels TGMF, van Norren D (1995) Ultraviolet and green light cause different types of damage in rat retina. Invest Ophthalmol Vis Sci 36:851–863Google Scholar
  9. Hertz H (1881) Über die Berührung fester elastischer Körper. Journal für die reine und angewandte. Mathematik 92:156–171zbMATHGoogle Scholar
  10. Kaunas R, Nguyen P, Usami S, Chien S (2005) Cooperative effects of Rho and mechanical stretch on stress fiber organization. Proc Natl Acad Sci USA 102:15895–15900CrossRefGoogle Scholar
  11. Kaunas R, Hsu H, Deguchi S (2011) Sarcomeric model of stretch-induced stress fiber reorganization. Cell Health Cytoskelet 3:13–22Google Scholar
  12. Keilbassa C, Roza L, Epe B (1997) Wavelength dependence of oxidative DNA damage induced by UV and visible light. Carcinogenesis 18:811–816CrossRefGoogle Scholar
  13. King M, Drivas T, Blobel G (2008) A network of nuclear envelope membrane proteins linking centromeres to microtubules. Cell 134:427–438CrossRefGoogle Scholar
  14. Krisch RE, Flick MB, Trumbore CN (1991) Radiation chemical mechanisms of single- and double-strand break formation in irradiated SV40 DNA. Radiat Res 126(2):251–259CrossRefGoogle Scholar
  15. Lee E, Kim DY, Chung E, Lee EA, Park KS, Son Y (2014) Transplantation of cyclic stretched fibroblasts accelerates the wound-healing process in streptozotocin-induced diabetic mice. Cell Transpl 23(3):285–301CrossRefGoogle Scholar
  16. Li B, Lin M, Tang Y, Wang B, Wang JH (2008) A novel functional assessment of the differentiation of micropatterned muscle cells. J Biomech 41(16):3349–3353CrossRefGoogle Scholar
  17. Luxton GW, Gomes ER, Folker ES, Vintinner E, Gundersen GG (2010) Linear arrays of nuclear envelope proteins harness retrograde actin flow for nuclear movement. Science 329(5994):956–959CrossRefGoogle Scholar
  18. Martens JC, Radmacher M (2008) Softening of the actin cytoskeleton by inhibition of myosin II. Pflugers Arch Eur J Physiol 456:95–100CrossRefGoogle Scholar
  19. Na S, Trache A, Trzeciakowski J, Sun Z, Meininger GA, Humphrey JD (2008) Time-dependent changes in smooth muscle cell stiffness and focal adhesion area in response to cyclic equibiaxial stretch. Ann Biomed Eng 36:369–380CrossRefGoogle Scholar
  20. Nagayama K (2015) Quantitative analysis of cellular traction forces using a micropillar substrate and estimation of the intracellular force applied to the nucleus. Trans Jpn Soc Mech Eng 81:824 (in Japanese) Google Scholar
  21. Nagayama K, Yahiro Y, Matsumoto T (2011) Stress fibers stabilize the position of intranuclear DNA through mechanical connection with the nucleus in vascular smooth muscle cells. FEBS Lett 585(24):3992–3997CrossRefGoogle Scholar
  22. Nagayama K, Kimura Y, Makino N, Matsumoto T (2012) Strain waveform dependence of stress fiber reorientation in cyclically stretched osteoblastic cells: effects of viscoelastic compression of stress fibers. Am J Physiol Cell Physiol 302(10):1469–1478CrossRefGoogle Scholar
  23. Nagayama K, Yamazaki S, Yahiro Y, Matsumoto T (2014) Estimation of the mechanical connection between apical stress fibers and the nucleus in vascular smooth muscle cells cultured on a substrate. J Biomech 47(6):1422–1429CrossRefGoogle Scholar
  24. Nishimura K, Li W, Hoshino Y, Kadohama T, Asada H, Ohgi S, Sumpio BE (2006) Role of AKT in cyclic strain-induced endothelial cell proliferation and survival. Am J Physiol Cell Physiol 290(3):C812–C821CrossRefGoogle Scholar
  25. Rabinovitz I, Gipson IK, Mercurio AM (2001) Traction forces mediated by alpha6beta4 integrin: implications for basement membrane organization and tumor invasion. Mol Biol Cell 12(12):4030–4043CrossRefGoogle Scholar
  26. Reme C, Reinboth J, Clausen M, Hafezi F (1996) Light damage revisited: converging evidence, diverging views? Graefes Arch Clin Exp Ophthalmol 234:2–11CrossRefGoogle Scholar
  27. Rozanowska M, Jarvisevans J, Korytowski W et al (1995) Blue lightinduced reactivity of retinal age pigment: in vitro generation of oxygen-reactive species. J Biol Chem 270:18825–18830CrossRefGoogle Scholar
  28. Shaul Y, Ben-Yehoyada M (2005) Role of c-Abl in the DNA damage stress response. Cell Res 15(1):33–35CrossRefGoogle Scholar
  29. Stary A, Robert C, Sarasin A (1997) Deleterious effects of ultraviolet A radiation in human cells. Mutat Res 383:1–8CrossRefGoogle Scholar
  30. Takata H, Hanafusa T, Mori T, Shimura M, Iida Y, Ishikawa K, Yoshikawa K, Yoshikawa Y, Maeshima K (2013) Chromatin compaction protects genomic DNA from radiation damage. PLoS ONE 8(10):e75622CrossRefGoogle Scholar
  31. Takemasa T, Yamaguchi T, Yamamoto Y, Sugimoto K, Yamashita K (1998) Oblique alignment of stress fibers in cells reduces the mechanical stress in cyclically deforming fields. Eur J Cell Biol 77:91–99CrossRefGoogle Scholar
  32. Versaevel M, Grevesse T, Gabriele S (2012) Spatial coordination between cell and nuclear shape within micropatterned endothelial cells. Nat Commun 3:671CrossRefGoogle Scholar
  33. Wang N, Tytell J, Ingber D (2009) Mechanotransduction at a distance: mechanically coupling the extracellular matrix with the nucleus. Nat Rev Mol Cell Biol 10:75–82CrossRefGoogle Scholar
  34. Xiong H, Rivero F, Euteneuer U, Mondal S, Mana-Capelli S, Larochelle D, Vogel A, Gassen B, Noegel AA (2008) Dictyostelium Sun-1 connects the centrosome to chromatin and ensures genome stability. Traffic 9(5):708–724CrossRefGoogle Scholar
  35. Yano S, Komine M, Fujimoto M, Okochi H, Tamaki K (2004) Mechanical stretching in vitro regulates signal transduction pathways and cellular proliferation in human epidermal keratinocytes. J Invest Dermatol 122(3):783–790CrossRefGoogle Scholar
  36. Zeng B, Tong S, Ren X, Xia H (2016) Cardiac cell proliferation assessed by EdU, a novel analysis of cardiac regeneration. Cytotechnology 68(4):763–770CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Micro-Nano Biomechanics Laboratory, Department of Mechanical Systems EngineeringIbaraki UniversityHitachiJapan

Personalised recommendations