Journal of Neuro-Oncology

, Volume 104, Issue 3, pp 697–704 | Cite as

Role of sonic hedgehog signaling in migration of cell lines established from CD133-positive malignant glioma cells

  • Hiroyuki Uchida
  • Kazunori AritaEmail author
  • Shunji Yunoue
  • Hajime Yonezawa
  • Yoshinari Shinsato
  • Hiroto Kawano
  • Hirofumi Hirano
  • Ryosuke Hanaya
  • Hiroshi Tokimura
Laboratory Investigation - Human/Animal Tissue


The sonic hedgehog (SHH) signaling pathway is essential for normal development and embryogenic morphogenesis. In malignant neoplasms its inappropriate activation correlates with tumorigenesis, proliferation, and migration. However, the role of SHH in infiltrative growth of glioblastoma remains to be elucidated. CD133 is a marker of tumor stem cells in glioblastoma, which are thought to play important roles in tumorigenesis, drug resistance, and tumor recurrence. We investigated the role of the SHH signaling pathway in migration of glioblastoma cell lines derived from CD133-positive cells. Two cell lines, GBM1 and GBM2, were established from CD133-positive cells sorted on an automagnetic cell separator from dispersed human glioblastoma cells. Both cell lines exhibited sphere-like growth in serum-free medium containing growth factor. Expression of patched (PTCH)-, a receptor of SHH, of smoothened (SMO)-, a 7 transmembrane receptor, and of GLI1- and GLI2, PTCH cascade signal proteins, was evaluated by reverse-transcription polymerase chain reaction (RT-PCR). The effects of recombinant SHH in the medium, and of knockdown of SMO-, GLI1- or GLI2 messenger RNA (mRNA) on the migratory ability of neoplastic cells were evaluated by scratch assays. RT-PCR revealed the presence of PTCH-, SMO-, GLI1-, and GLI2 mRNA in these cells. Their migratory ability was significantly enhanced (P < 0.05) by addition of recombinant SHH to the medium. Knockdown of SMO-, GLI1- or GLI2 mRNA resulted in significant decrease in the mobility of the neoplastic cells. Our study suggests that the SHH pathway plays an important role in the migratory ability of cells derived from CD133-positive human glioblastoma cells.


Sonic hedgehog pathway Glioblastoma CD133-positive cells Migration 



This work was aided by research grants, Kiban Research Grant C (to K.A.) and Wakate Research Grant (to S.Y.) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan. No conflicts of interests prejudicing the impartiality of this paper are declared.

Supplementary material

11060_2011_552_MOESM1_ESM.pdf (20 kb)
Supplementary figure - Sufficient depletion in the expression of mRNA was confirmed in mRNA-silenced cells. (PDF 20 kb)


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Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Hiroyuki Uchida
    • 1
  • Kazunori Arita
    • 1
    Email author
  • Shunji Yunoue
    • 1
  • Hajime Yonezawa
    • 1
  • Yoshinari Shinsato
    • 1
  • Hiroto Kawano
    • 1
  • Hirofumi Hirano
    • 1
  • Ryosuke Hanaya
    • 1
  • Hiroshi Tokimura
    • 1
  1. 1.Department of NeurosurgeryKagoshima University Graduate School of Medical and Dental SciencesKagoshimaJapan

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