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Higher cell stiffness indicating lower metastatic potential in B16 melanoma cell variants and in (−)-epigallocatechin gallate-treated cells

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Abstract

Purpose

To understand how nanomechanical stiffness affects metastatic potential, we studied the relationship between cell migration, a characteristic of metastasis, and cell stiffness using atomic force microscopy (AFM), which can measure stiffness (elasticity) of individual living cells.

Methods

Migration and cell stiffness of three metastatic B16 melanoma variants (B16-F10, B16-BL6, and B16-F1 cells), and also effects of (−)-epigallocatechin gallate (EGCG), were studied using Transwell assay and AFM.

Results

Migration of B16-F10 and B16-BL6 cells was 3 and 2 times higher than that of B16-F1 cells in Transwell assay, and cell stiffness determined by AFM was also different among the three variants, although they have similar morphologies and the same growth rates: Means of Young’s modulus were 350.8 ± 4.8 Pa for B16-F10 cells, 661.9 ± 16.5 Pa for B16-BL6 cells, and 727.2 ± 13.0 Pa for B16-F1 cells. AFM measurements revealed that highly motile B16-F10 cells have low cell stiffness, and low motile and metastatic B16-F1 cells have high cell stiffness: Nanomechanical stiffness is inversely correlated with migration potential. Treatment of highly motile B16-F10 cells with EGCG increased cell stiffness 2-fold and inhibited migration of the cells.

Conclusions

Our study with AFM clearly demonstrates that cell stiffness is a reliable quantitative indicator of migration potential, and very likely metastatic potential, even in morphologically similar cells. And increased cell stiffness may be a key nanomechanical feature in inhibition of metastasis.

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Abbreviations

AFM:

Atomic force microscopy

EGCG:

(−)-Epigallocatechin gallate

MβCD:

Methyl-β-cyclodextrin

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Acknowledgments

We thank Drs. James K. Gimzewski and Shivani Sharma, University of California, and Drs. Kunihiko Okajima and Mariko Ago, Tokushima Bunri University, for their fruitful discussion. We also thank Dr. Shun’ichiro Taniguchi for the supply of B16 variants, and Mr. Takahisa Matsuzaki, Ms. Kaori Suzuki and Ikuko Shiotani, Research Institute for Clinical Oncology, Saitama Cancer Center, for their technical assistance. This work was supported by the Smoking Research Fundation, Urakami Foundation, and World Premier International Research Center Initiative on Materials Nanoarchitectonics (H. K. and T. N.). This work was supported in part by the Nanotechnology Network Japan Program from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Conflict of interest

The authors did not have any conflicts of interest to disclose.

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Authors and Affiliations

  1. Research Institute for Clinical Oncology, Saitama Cancer Center, Kitaadachi-gun, Saitama, 362-0806, Japan

    Tatsuro Watanabe, Atsushi Takahashi & Masami Suganuma

  2. National Institute for Materials Science, International Center for Materials Nanoarchitectonics, Ibaraki, Japan

    Hiromi Kuramochi & Tomonobu Nakayama

  3. Graduate School of Science and Engineering, Saitama University, Sakura-ku, Saitama, Japan

    Atsushi Takahashi

  4. Department of Radiobiology and Molecular Epidemiology, Radiation Effects Research Foundation, Hiroshima, Japan

    Kazue Imai

  5. Nanotechnology Innovation Center, National Institute for Materials Science, Ibaraki, Japan

    Naoko Katsuta

  6. Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Tokushima, Japan

    Hirota Fujiki

Authors
  1. Tatsuro Watanabe
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  2. Hiromi Kuramochi
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  3. Atsushi Takahashi
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  4. Kazue Imai
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  6. Tomonobu Nakayama
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  7. Hirota Fujiki
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  8. Masami Suganuma
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Corresponding author

Correspondence to Masami Suganuma.

Additional information

T. Watanabe and H. Kuramochi contributed equally to this work.

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Watanabe, T., Kuramochi, H., Takahashi, A. et al. Higher cell stiffness indicating lower metastatic potential in B16 melanoma cell variants and in (−)-epigallocatechin gallate-treated cells. J Cancer Res Clin Oncol 138, 859–866 (2012). https://doi.org/10.1007/s00432-012-1159-5

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  • DOI: https://doi.org/10.1007/s00432-012-1159-5

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