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Clinical & Experimental Metastasis

, Volume 27, Issue 8, pp 619–630 | Cite as

Mesenchymal mode of migration participates in pulmonary metastasis of mouse osteosarcoma LM8

  • Yoshihiro YuiEmail author
  • Kazuyuki Itoh
  • Kiyoko Yoshioka
  • Norifumi Naka
  • Motonobu Watanabe
  • Yoshimi Hiraumi
  • Hiroshi Matsubara
  • Ken-ichiro Watanabe
  • Kazumi Sano
  • Tatsutoshi Nakahata
  • Souichi Adachi
Research Paper

Abstract

The outcomes of osteosarcoma patients still remain poor because of intractable pulmonary metastasis. We previously established a highly metastatic osteosarcoma cell line, LM8 from Dunn mouse osteosarcoma by in vivo selection. We herein aimed to clarify the characteristic biological features related with high metastatic potential and new target molecules to suppress pulmonary metastasis of osteosarcoma, using this syngeneic spontaneous metastatic model. LM8 cells acquired fibroblastic morphology with striking filopodia on the cell surface. Immunostaining showed faint stress fiber formation and peripherally localized integrin β1, and biochemical analyses showed the activated Cdc42 and autophosphorylation of focal adhesion kinase (FAK) in LM8 cells when compared to Dunn cells. LM8 cells had activated motility in single cell migration mode. LM8 migration was increased by a Rho-associated kinase (ROCK) inhibitor, Y-27632, while decreased by Cdc42 silencing using RNA interference system. We found that a clinically approved camptothecin analog, irinotecan suppressed the migration, Cdc42 activity, and autophosphorylation of FAK, and attenuated integrin β1 distribution selectively in LM8 cells. Daily oral administration of irinotecan significantly reduced the rate and size of pulmonary metastasis in syngeneic C3H mice. The fibroblastic morphology and activated cell migration with the dependency on Cdc42 but not Rho-ROCK signaling pathway argued that LM8 moved in mesenchymal mode of cell migration. This activated mesenchymal migration was a key component of the pulmonary metastasis of LM8 cells. The inhibition of mesenchymal migration by irinotecan, in addition to its cytotoxic effects, might be effective in preventing pulmonary metastasis of osteosarcoma.

Keywords

Osteosarcoma Metastasis Mesenchymal migration Cdc42 Irinotecan 

Abbreviations

ECM

Extracellular matrix

3D

Three dimensions

FAK

Focal adhesion kinase

MMP

Matrix metalloproteinase

ROCK

Rho-associated kinase

Notes

Acknowledgments

We thank Dr. Takahashi (The 21st Century COE Formation, Kyoto University Graduate School of Medicine, Japan) for the expressing vectors of GST-Rhotekin and GST-PAK, and Dr. Yukawa (GE Healthcare Bioscience, Tokyo, Japan) for the analysis of EZ-TAXIScan data.

Supplementary material

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Yoshihiro Yui
    • 1
    • 2
    Email author
  • Kazuyuki Itoh
    • 2
  • Kiyoko Yoshioka
    • 2
  • Norifumi Naka
    • 3
  • Motonobu Watanabe
    • 1
  • Yoshimi Hiraumi
    • 1
  • Hiroshi Matsubara
    • 1
  • Ken-ichiro Watanabe
    • 1
  • Kazumi Sano
    • 4
  • Tatsutoshi Nakahata
    • 1
  • Souichi Adachi
    • 1
  1. 1.Department of PediatricsKyoto University Graduate School of MedicineKyotoJapan
  2. 2.Department of BiologyOsaka Medical Center of Cancer and Cardiovascular DiseasesOsakaJapan
  3. 3.Muscloskeletal Oncology ServiceOsaka Medical Center of Cancer and Cardiovascular DiseasesOsakaJapan
  4. 4.Department of Drug Metabolism and DispositionMeiji Pharmaceutical UniversityKiyoseJapan

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