Skip to main content

Advertisement

SpringerLink
Log in

Adenovirus-mediated transfer of siRNA against basic fibroblast growth factor mRNA enhances the sensitivity of glioblastoma cells to chemotherapy

  • Original Paper
  • Published:
Medical Oncology Aims and scope Submit manuscript

Abstract

Basic fibroblast growth factor (bFGF) is an important growth factor for glioma cell proliferation and invasion. BFGF is overexpressed in malignant gliomas and its level is associated with malignant grades and clinical outcome of patients with gliomas. Small interfering RNAs (siRNA) are synthetic forms of microRNA made of short double stranded RNA, and they effectively catalyze the degradation of their target mRNA. The use of siRNA has become a key method in the suppression of gene expression and the development of therapeutic agents. In this study, we used an adenovirus(Ad)-mediated transfer of siRNA against bFGF mRNA (Ad-bFGF-siRNA) to study the effect of down-regulating bFGF expression on the sensitivity of glioma cells to chemotherapeutics. An optimal siRNA sequence specific for bFGF mRNA was cloned into an adenoviral vector and transfected into three glioma cell lines: U251, A172, and LN229. Methyl thiazolyl tetrazolium (MTT) assays were used to examine changes in cell proliferation, and changes in bFGF mRNA and protein levels in U251 cells were detected using quantitative RT-PCR and Western blot, respectively. Apoptosis of U251 cells was detected using Hoechst staining and flow cytometry, with expression of apoptosis-related proteins evaluated by Western blot. Following the transfection of a bFGF-specific siRNA, mRNA and protein levels of bFGF decreased significantly. Lower rates of proliferation and increased levels of apoptosis also were associated with the Ad-bFGF-siRNA-transfected group compared to control group. Decreased expression of Bcl-2, Bcl-xL, Jak-1, and STAT-3 and increased expression of Bax also were detected in the Ad-bFGF-siRNA-transfected group. For cells treated with both Ad-bFGF-siRNA and chemotherapeutics, a significant reduction in cell survival was observed compared to treatment with Ad-bFGF-siRNA or chemotherapeutics alone. Overall, we found that targeting bFGF mRNA with a siRNA resulted in lower rates of proliferation, increased apoptosis, and enhanced sensitivity of glioma cells to chemotherapy drugs. This suggests that specific targeting of bFGF mRNA may provide a novel approach for the treatment of glioblastoma multiforme (GBM).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

Ad:

Adenovirus

bFGF:

Basic fibroblast growth factor

GBM:

Glioblastoma multiforme

GFP:

Green fluorescent protein

MOI:

Multiplicity of infection

MTT:

3-(4,5)-dimethylthiahiazo(-2-y1)-2,5-di- phenytetrazoliumromide

STAT:

Signal transduction and activator of transcription

BCNU:

1,3-bis(2-chloroethyl)-1-nitrosourea

VM-26:

Teniposide

References

  1. Wen PY, Kesari s. Malignant gliomas in adults. N Engl J Med. 2008;359:492–507.

    Article  PubMed  CAS  Google Scholar 

  2. Grzelinski M, Urban-Klein B, Martens T, Lamszus K, Bakowsky U, Hobel S, et al. RNA interference-mediated gene silencing of pleiotrophin through polyethylenimine-complexed small interfering RNAs in vivo exerts antitumoral effects in glioblastoma xenografts. Hum Gene Ther. 2006;17:751–66.

    Article  PubMed  CAS  Google Scholar 

  3. Stegh AH, Kim H, Bachoo RM, Forloney KL, Zhang J, Schulze H, et al. Bcl2L12 inhibits post-mitochondrial apoptosis signaling in glioblastoma. Genes Dev. 2007;21:98–111.

    Article  PubMed  CAS  Google Scholar 

  4. Kim HS. Assignment1 of the human basic fibroblast growth factor gene FGF2 to chromosome 4 band q26 by radiation hybrid mapping. Cytogenet Cell Genet. 1998;83:73.

    Article  PubMed  CAS  Google Scholar 

  5. Eckenstein FP. Fibroblast growth factors in the nervous system. J Neurobiol. 1994;25:1467–80.

    Article  PubMed  CAS  Google Scholar 

  6. Fukui S, Nawashiro H, Otani N, Ooigawa H, Nomura N, Yano A, et al. Nuclear accumulation of basic fibroblast growth factor in human astrocytic tumors. Cancer. 2003;97:3061–7.

    Article  PubMed  CAS  Google Scholar 

  7. Baguma-Nibasheka M, Li AW, Murphy PR. The fibroblast growth factor-2 antisense gene inhibits nuclear accumulation of FGF-2 and delays cell cycle progression in C6 glioma cells. Mol Cell Endocrinol. 2007;267:127–36.

    Article  PubMed  CAS  Google Scholar 

  8. Bikfalvi A, Klein S, Pintucci G, Rifkin DB. Biological roles of fibroblast growth factor-2. Endocr Rev. 1997;18:26–45.

    Article  PubMed  CAS  Google Scholar 

  9. Aoki T, Kato S, Fox JC, Okamoto K, Sakata K, Morimatsu M, et al. Inhibition of autocrine fibroblast growth factor signaling by the adenovirus-mediated expression of an antisense transgene or a dominant negative receptor in human glioma cells in vitro. Int J Oncol. 2002;21:629–36.

    PubMed  CAS  Google Scholar 

  10. Takahashi JA, Fukumoto M, Kozai Y, Ito N, Oda Y, Kikuchi H, et al. Inhibition of cell growth and tumorigenesis of human glioblastoma cells by a neutralizing antibody against human basic fibroblast growth factor. FEBS Lett. 1991;288:65–71.

    Article  PubMed  CAS  Google Scholar 

  11. Kim D, Rossi J. RNAi mechanisms and applications. Biotechniques. 2008;44:613–6.

    Article  PubMed  CAS  Google Scholar 

  12. Wang SJ, Wang JH, Zhang YW, Xu XN, Liu HS. Effects of small interfering RNA targeting basic fibroblast growth factor on proliferation and apoptosis of glioma cell line U251. Ai Zheng. 2008;27:905–9.

    PubMed  CAS  Google Scholar 

  13. Shen C, Buck AK, Liu X, Winkler M, Reske SN. Gene silencing by adenovirus-delivered siRNA. FEBS Lett. 2003;539:111–4.

    Article  PubMed  CAS  Google Scholar 

  14. Zhao YP, Bai WJ, Yan Z, Chen GQ, Liu SJ, Deng QP, et al. Construction of an antisense RNA recombinant adenovirus vector of the human PDE5A1 gene promoter. Zhonghua Nan Ke Xue. 2005;11:331–4.

    PubMed  Google Scholar 

  15. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25:402–8.

    Article  PubMed  CAS  Google Scholar 

  16. Noda S, Yoshimura S, Sawada M, Naganawa T, Iwama T, Nakashima S, et al. Role of ceramide during cisplatin-induced apoptosis in C6 glioma cells. J Neurooncol. 2001;52:11–21.

    Article  PubMed  CAS  Google Scholar 

  17. He L, Kim BY, Kim KA, Kwon O, Kim SO, Bae EY, et al. NF-kappaB inhibition enhances caspase-3 degradation of Akt1 and apoptosis in response to camptothecin. Cell Signal. 2007;19:1713–21.

    Article  PubMed  CAS  Google Scholar 

  18. Bian XW, Du LL, Shi JQ, Cheng YS, Liu FX. Correlation of bFGF, FGFR-1 and VEGF expression with vascularity and malignancy of human astrocytomas. Anal Quant Cytol Histol. 2000;22:267–74.

    PubMed  CAS  Google Scholar 

  19. Auguste P, Gursel DB, Lemiere S, Reimers D, Cuevas P, Carceller F, et al. Inhibition of fibroblast growth factor/fibroblast growth factor receptor activity in glioma cells impedes tumor growth by both angiogenesis-dependent and -independent mechanisms. Cancer Res. 2001;61:1717–26.

    PubMed  CAS  Google Scholar 

  20. De-Fraja C, Conti L, Govoni S, Battaini F, Cattaneo E. STAT signalling in the mature and aging brain. Int J Dev Neurosci. 2000;18:439–46.

    Article  PubMed  CAS  Google Scholar 

  21. Zeng ZZ, Yellaturu CR, Neeli I, Rao GN. 5(S)-hydroxyeicosatetraenoic acid stimulates DNA synthesis in human microvascular endothelial cells via activation of Jak/STAT and phosphatidylinositol 3-kinase/Akt signaling, leading to induction of expression of basic fibroblast growth factor 2. J Biol Chem. 2002;277:41213–9.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Sciences Foundation of China (30672158).

Author information

Authors and Affiliations

  1. Department of Neurosurgery, Tianjin First Center Hospital, Fukang Road No. 24, 300192, Tianjin, China

    Xuequan Feng & Jinhuan Wang

  2. Key Lab for Critical Care Medicine of the Ministry of Health, Tianjin First Center Hospital, Tianjin, China

    Biao Zhang, Jinhuan Wang, Xinnv Xu, Na Lin & Hongsheng Liu

Authors
  1. Xuequan Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Biao Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jinhuan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xinnv Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Na Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hongsheng Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jinhuan Wang.

Additional information

X. Feng, B. Zhang contributed equally to this work and should be considered as co-first authors.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Feng, X., Zhang, B., Wang, J. et al. Adenovirus-mediated transfer of siRNA against basic fibroblast growth factor mRNA enhances the sensitivity of glioblastoma cells to chemotherapy. Med Oncol 28, 24–30 (2011). https://doi.org/10.1007/s12032-010-9445-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12032-010-9445-z

Keywords

Search

Navigation