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Journal of Neuro-Oncology

, Volume 93, Issue 2, pp 183–189 | Cite as

c-Met-targeted RNA interference inhibits growth and metastasis of glioma U251 cells in vitro

  • Sheng-Hua Chu
  • Dong-Fu Feng
  • Hong Zhang
  • Er-Tao Chen
  • Zhi-Xin Duan
  • Xue-Yuan Li
  • Jia Li
  • Yan-Bin MaEmail author
  • Zhi-An Zhu
  • Jian-Hua Qiu
Laboratory investigation - human/animal tissue

Abstract

Angiogenesis plays an essential role in tumor growth and metastasis and is a promising target for cancer therapy. c-Met, a receptor tyrosine kinase, and its ligand, hepatocyte growth factor (HGF), are critical in cellular proliferation, motility, invasion, and angiogenesis. The present study was designed to determine the role of c-Met in growth and metastasis of glioma U251 cells using RNA interference (RNAi) technology in vitro. We constructed three kinds of shRNA expression vectors aiming at the c-Met gene, then transfected them into glioma U251 cells by lipofectamineTM 2000. The level of c-Met mRNA was investigated by real-time polymerse chain reaction (RT-PCR). The protein expression of c-Met was observed by immunofluoresence staining and western blotting. U251 cell growth and adherence was detected by methyl thiazole tetrazolium assay. The apoptosis of U251 cells was examined with a flow cytometer. The adherence, invasion, and in vitro angiogenesis assays of U251 cells were done. We got three kinds of c-Met specific shRNA expression vectors which could efficiently inhibit the growth and metastasis of U251 cells and the expression of c-Met in U251 cells. RT-PCR, immunofluoresence staining and western blotting showed that inhibition rate for c-Met expression was up to 90%, 79% and 85%, respectively. The expression of c-Met can be inhibited by RNA interference in U251 cells, which can inhibit the growth and metastasis of U251 cell and induce cell apoptosis. These results indicate that RNAi of c-Met can be an effective antiangiogenic strategy for glioma.

Keywords

c-Met Apoptosis Adherence Gene therapy Glioma RNA interference 

Abbreviations

HGF

Hepatocyte growth factor

c-Met

Hepatocyte growth factor receptor

RNAi

RNA interference

RPMI

Roswell park memorial institute

PBS

Phosphate-buffered saline

SDS-PAGE

Sodium dodecyl sulphate polyacrylamide gel electrophoresis

HRP

Horseradish peroxidase

NC

Nitrocellulose

MTT

Methyl thiazole tetrazolium

Notes

Acknowledgements

We thank professor Hong Wang at Henry Ford Hospital for critical reading of the paper. This work was supported by a grant 07JWYQ03 from the Training Excellent Youth Teacher Scientific Research Foundation of University of Shanghai, and a grant 07XYQ01 from the Excellent Youth Teacher Scientific Research Foundation of Shanghai Jiao Tong University of School of Medicine.

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Copyright information

© Springer Science+Business Media, LLC. 2009

Authors and Affiliations

  • Sheng-Hua Chu
    • 1
  • Dong-Fu Feng
    • 1
  • Hong Zhang
    • 1
  • Er-Tao Chen
    • 1
  • Zhi-Xin Duan
    • 1
  • Xue-Yuan Li
    • 1
  • Jia Li
    • 1
  • Yan-Bin Ma
    • 1
    Email author
  • Zhi-An Zhu
    • 1
  • Jian-Hua Qiu
    • 1
  1. 1.Department of NeurosurgeryNo. 3 People’s Hospital Affiliated to Shanghai Jiao Tong University School of MedicineShanghaiChina

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