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

  • Laboratory investigation - human/animal tissue
  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

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.

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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

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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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Authors and Affiliations

  1. Department of Neurosurgery, No. 3 People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 201900, China

    Sheng-Hua Chu, Dong-Fu Feng, Hong Zhang, Er-Tao Chen, Zhi-Xin Duan, Xue-Yuan Li, Jia Li, Yan-Bin Ma, Zhi-An Zhu & Jian-Hua Qiu

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  1. Sheng-Hua Chu
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Correspondence to Yan-Bin Ma.

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Chu, SH., Feng, DF., Zhang, H. et al. c-Met-targeted RNA interference inhibits growth and metastasis of glioma U251 cells in vitro. J Neurooncol 93, 183–189 (2009). https://doi.org/10.1007/s11060-008-9772-5

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  • DOI: https://doi.org/10.1007/s11060-008-9772-5

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