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Journal of Neuro-Oncology

, Volume 103, Issue 2, pp 267–276 | Cite as

Downregulation of uPARAP mediates cytoskeletal rearrangements and decreases invasion and migration properties in glioma cells

  • Satoshi Takahashi
  • Hisafumi Yamada-Okabe
  • Kenji Hamada
  • Shigeki Ohta
  • Takeshi Kawase
  • Kazunari Yoshida
  • Masahiro TodaEmail author
Laboratory Investigation - Human/Animal Tissue

Abstract

To identify molecular therapeutic targets for glioma, we performed gene expression profiling by using a complementary DNA (cDNA) microarray method and identified the urokinase plasminogen activator receptor-associated protein (uPARAP/Endo180) as a gene expressed highly in glioma tissue compared with the normal brain tissue. The uPARAP is an endocytic receptor for collagen. In certain cell types, uPARAP occurs in a complex with the urokinase plasminogen activator receptor (uPAR) where it fulfills other functions in addition to collagenolysis. Quantitative PCR analysis using a cDNA panel revealed higher expression levels of uPARAP in glioma tissue compared with normal brain tissue. Western blot analysis revealed that the uPARAP protein was expressed in glioma samples and two glioma cell lines, KNS42 and KNS81, but not expressed in control tissue from the normal brain. Introduction of small interfering RNA-targeted uPARAP into the two different glioma cell lines, KNS42 and KNS81, resulted in downregulation of uPARAP expression, and it significantly suppressed glioma cell migration and invasion in vitro. Control glioma cells showed small cell bodies, whereas uPARAP siRNA-treated glioma cells exhibited large and flat morphology. Most of the polymeric actin in the control glioma cells was concentrated in the lamellipodia that are observed in mobile cells. In contrast, in the uPARAP siRNA-treated glioma cells, polymeric actin became organized in stress fibers and the lamellipodia disappeared, characteristic of immobile cells. Our present study suggests that uPARAP may be involved in glioma cell invasiveness through actin cytoskeletal rearrangement. downregulation of uPARAP may be a novel anti-invasion therapeutic strategy for malignant gliomas.

Keywords

Glioma invasion Migration uPARAP Endo 180 Actin cytoskeleton 

Notes

Acknowledgments

We thank Ms. S. Teramoto, Ms. T. Muraki and Ms. Y. Aikawa for technical assistance, and Dr. R. Fukaya for useful discussion. Grants from the Ministry of Education, Science, Sports, and Science and Technology, JAPAN including Grant-in-Aid for Young Scientists (B) (JSPS KAKENHI 19791011).

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Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Satoshi Takahashi
    • 1
  • Hisafumi Yamada-Okabe
    • 2
  • Kenji Hamada
    • 3
  • Shigeki Ohta
    • 4
  • Takeshi Kawase
    • 1
  • Kazunari Yoshida
    • 1
  • Masahiro Toda
    • 1
    Email author
  1. 1.Department of Neurosurgery, School of MedicineKeio UniversityTokyoJapan
  2. 2.Research Planning and Coordination Department, Gotenba Research LaboratoriesChugai Pharmaceutical Co., Ltd.ShizuokaJapan
  3. 3.Pharmaceutical Research Department, Kamakura Research LaboratoriesChugai Pharmaceutical Co., Ltd.KanagawaJapan
  4. 4.Division of Cellular Signaling, Institute for Advanced Medical Research, School of MedicineKeio UniversityTokyoJapan

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