Advertisement

Silencing Epidermal Growth Factor Receptor by RNA Interference in Glioma

  • Chunsheng Kang
  • Peiyu Pu
  • Hao Jiang
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 542)

Summary

Glioblastoma multiforme (GBM) can arise de novo or progress from a lower to higher grade and can possess a series of genetic alterations and dynamic progressions, which have been correlated with the molecular pathology of GBM. Epidermal growth factor receptor (EGFR) has been shown to be overexpressed in a variety of tumors and is one of the important mediators responsible for the development of high-grade gliomas, especially in primary glioblastomas. Most recently, RNA interference (RNAi), in which double-stranded RNA (dsRNA) induces sequence-specific degradation of the targeting messenger RNA (mRNA), has been extensively developed and studied. RNAi is able to silence the targeted gene expression more efficiently and specifically. In the present study, we silence the EGFR expression using two separate short interfering RNAs (siRNAs) targeting the extracellular ligand-binding domain and intracellular tyrosine kinase domain, respectively. We demonstrate that suppression of EGFR expression, by using either antisense or siRNA approaches, inhibits U251 glioblastoma cell growth in vitro and in vivo, and siRNA seems to be more effective than the antisense approach.

Keywords

EGFR glioma invasion proliferation siRNA 

Notes

Acknowledgments

This work is supported by Tianjin Science and Technology Committee, grant numbers 05YFJZJC1002 and 06YFSZSF01100; the China National Natural Scientific Found (30772231); and the Program for New Century Excellent Talents in University, The Ministry of Education of the People`s Republic of China (grant number NCET-07-0615).

References

  1. 1.
    Stewart LA. (2002) Chemotherapy in adult high-grade glioma: a systematic review and meta-analysis of individual patient data from 12 randomised trials. Lancet . 359, 1011 –1018.PubMedCrossRefGoogle Scholar
  2. 2.
    Mendelsohn J, Baselga J. (2000) The EGF receptor family as targets for cancer therapy. Oncogene . 19, 6550 –6665.PubMedCrossRefGoogle Scholar
  3. 3.
    Wells A. (1999) EGF receptor. Review. Int J Biochem Cell . 31, 637 –643.CrossRefGoogle Scholar
  4. 4.
    Pu P, Liu X, Liu A, Cui J, Zhang Y. (2000) Inhibitory effect of antisense epidermal growth factor receptor RNA on the proliferation of rat C6 glioma cells in vitro and in vivo. J Neurosurg . 92, 132 –139.PubMedCrossRefGoogle Scholar
  5. 5.
    Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature . 391, 806 –811.PubMedCrossRefGoogle Scholar
  6. 6.
    Zamore PD. (2001) RNA interference: listening to the sound of silence. Nat Struct Biol . 8, 746 –751.PubMedCrossRefGoogle Scholar
  7. 7.
    Sijen T, Fleenor J, Simmer F, Thijssen KL, Parrish S, Timmons L, Plasterk RH, Fire A. (2001) On the role of RNA amplification in dsRNA-triggered gene silencing. Cell . 107, 465 –476.PubMedCrossRefGoogle Scholar
  8. 8.
    Moroni MC, Willingham MC, Beguinot L. (1992) EGFR antisense RNA blocks expression of epidermal growth factor receptor and suppresses the transforming phenotype of a human carcinoma cell line. J Biol Chem . 267, 2714 –2722.PubMedGoogle Scholar
  9. 9.
    Ui-Tei K, Naito Y, Takahashi F, Haraguchi T, Ohki-Hamazaki H, Juni A, Ueda R, Saigo K. (2004) Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference. Nucleic Acids Res . 32, 936 –948.PubMedCrossRefGoogle Scholar
  10. 10.
    Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Webe K, Tuschl T. (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in mammalian cell culture. Nature . 411, 494 –498.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Chunsheng Kang
    • 1
  • Peiyu Pu
    • 1
  • Hao Jiang
    • 2
  1. 1.Department of NeurosurgeryTianjin Medical University General Hospital, Laboratory of Neuro-Oncology, Tianjin Neurological InstituteTianjinPeople’s Republic of China
  2. 2.Department of NeurologyHenry Ford HospitalDetroitUSA

Personalised recommendations