Skip to main content

Advertisement

SpringerLink
Log in

Antisense oligonucleodes targeting the focal adhesion kinase inhibit proliferation, induce apoptosis and cooperate with cytotoxic drugs in human glioma cells

  • Laboratory Investigation
  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Summary

To examine the role of focal adhesion kinase in human glioma cells, we studied its effects on proliferation and apoptosis using FAK antisense oligonucleotide. U251 MG cells were transfected with ODNs, sense FAK, mismatch FAK and antisense-FAK, respectively. Expression of FAK proteins were detected by Western blots and Immnofluoressence. Cell apoptosis and mitochondrial membrane potential were analyzed by flow cytometry. Caspase-3 activity was measured by spectrofluorometer. MTT assay was used to examine changes in cell proliferation. The protein expression of FAK in U251 MG cells decreased in antisense-FAK ODNs group significantly. Caspase-3 activity increased in cells treated with antisense-FAK and down-regulated when treated with caspase-3 inhibitor. The level of cell apoptosis and loss of mitochondrial membrane potential in antisense-FAK group was higher than in the mismatch sense group. Cells proliferation was inhibited by antisense-FAK, and the effects were clearly additive when antisense oligonuceotides were added to cells treated with the anticancer agents. The results suggest that antisense-FAK ODNs inhibit U251 MG cells proliferation and induce their apoptosis. It is possible that FAK via mitochondrial and caspase-3 inhibits U251 MG cells apoptosis. And antisense oligonucleotide treatment enhances U251 MG cells sensitivity to chemotherapy.

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

Similar content being viewed by others

References

  1. Reavey-Cantwill JF, Haroun RI, Zahurak M, Clatterbuck RE, Parker RJ, Mehta R, Fruehauf JP, Brem H., 2001 The prognostic value of tumor markers in patients with glioblastoma multiforme: analysis of 32 patients and review of the literature J Neuro-oncol 55:195–204

    Article  Google Scholar 

  2. Hauck CR, Hsia DA, Schlaepfer DD, 2002 The focal adhesion kinase – a regulator of cell migration and invasion IUBMB Life 53: 115–119

    Article  PubMed  CAS  Google Scholar 

  3. Owens LV, Xu L, Craven RJ, Dent GA, Weiner TM, Kornberg L, Liu ET, Cance WG, 1995 Overexpression of focal adhesion kinase (p125FAK) in invasive human tumors Cancer Res 55: 2752–2755

    PubMed  CAS  Google Scholar 

  4. Zagzag D, Friedlander DR, Margolis B, Grumet M, Semenza GL, Zhong H, Simons JW, Holash J, Wiegand SJ, Yancopoulos GD, 2000 Molecular events implicated in brain tumor angiogenesis and invasion Pediatr Neurosurg 33: 49–55

    Article  PubMed  CAS  Google Scholar 

  5. Jones G, Machado J Jr, Merlo A, 2001 Loss of focal adhesion kinase (FAK) inhibits epidermal growth factor receptor dependent migration and induces aggregation of NH2-terminal FAK in the nuclei of apoptotic glioblastoma cells Cancer Res 61:4978–4981

    PubMed  CAS  Google Scholar 

  6. Noda S, Yoshimura S, Sawada M, Naganawa T, Iwama T, Nakashima S, Sakai N, 2001 Role of ceremide during cisplatin-induced apoptosis in C6 glioma cells J Neuro-oncol 52: 11–21

    Article  CAS  Google Scholar 

  7. Minn AJ, Rudin CM, Boise LH, Thompson CB, 1995 Expression of bcl-xL can confer a multidrug resistance phenotype Blood 86(5): 1903–1910

    PubMed  CAS  Google Scholar 

  8. Sonoda Y, Matsumoto Y, Funakoshi M, Yamamoto D, Hanks SK, Kasahara T, 2000 Anti-apoptotic role of focal adhesion kinase (FAK) induction of inhibitor-of -apoptosis proteins and apoptosis suppression by the overexpression of FAK in human leukemic cell line, HL-60J Biol Chem 275: 16309–16315

    Article  PubMed  CAS  Google Scholar 

  9. Kim LT, Fleming JB, Lopez-Guzman C, Nwariaku F, 2003 Focal adhesions and associated proteins in medullary thyroid carcinoma cells J Surg Res 111: 177–184

    Article  PubMed  CAS  Google Scholar 

  10. Sasaki H, Kotsuji F, Tsang BK, 2002 Caspase 3-mediated focal adhesion kinase processing in human ovarian cancer cells: possible regulation by X-linked inhibitor of apoptosis protein Gynecol Oncol 85: 339–350

    Article  PubMed  CAS  Google Scholar 

  11. Alama A, Barbieri F, Cagnoli M, Schettini G, 1997 Antisense oligonuceotides as therapeutic agents Pharmacol Res 36: 171–178

    Article  PubMed  CAS  Google Scholar 

  12. Webb A, Cunningham D, Cotter F, Clarke PA, di Stefano F, Ross P, Corbo M, Dziewanowska Z, 1997 BCL-2 antisense therapy in patients with non-Hodgkin lymphoma Lancet 89: 988–990

    Google Scholar 

  13. Zhang R, Iyer RP, Yu D, Tan W, Zhang X, Lu Z, Zhao H, Agrawal S, 1996 Pharmacokinetics and tissue disposition of a chimeric oligodeoxynucleotide phosphorothioate in rat after intravenous administration J Pharmacol Exp Ther 278: 971–979

    PubMed  CAS  Google Scholar 

  14. Kronenwett R, Steidl U, Kirsch M, Sczakiel G, Haas G, 1998 Oligodeoxyribonucleotide uptake in primary human hepatopoietic cells is enhanced by cationic lipids and depends on the hepatopoietic cell subset Blood 91: 852–862

    PubMed  CAS  Google Scholar 

  15. Kasahara T, Koguchi E, Funakoshi M, Aizu-Yokota E, Sonoda Y, 2002 Antiapoptotic action of focal adhesion kinase (FAK) against ionizing radiation Antioxid Redox Signal 4: 491–499

    Article  PubMed  CAS  Google Scholar 

  16. Satoh TH, Surmacz TA, Nyormoi O, Whitacre CM, 2003 Inhibition of focal kinase by antisense oligonucleotides enhances the sensitivity of breast cancer cells to camptothecins Biocell 27: 47–55

    PubMed  CAS  Google Scholar 

  17. Blanc C, Deveraux QL, Krajewski S, Janicke RU, Porter AG, Reed JC, Jaggi R, Marti A, 2000 Caspase-3 is essential for procaspase-9 processing and cisplatin-induced apoptosis of MCF-7 breast cancer cells Cancer Res 60: 4386–4391

    PubMed  CAS  Google Scholar 

  18. Kothakota S, Azuma T, Reinhard C, Klippel A, Tang J, Chu K, McGarry TJ, Kirschner MW, Koths K, Kwiatkowski DJ, Williams LT, 1997 Caspase-3-generated fragment of gelsolin: effector of morphological change in apoptosis Science 278: 294–298

    Article  PubMed  CAS  Google Scholar 

  19. Takuma K, Lee E, Enomoto R, Baba A, Matsuda T, 2001 Ibudilast attenuates astrocyte apoptosis via cyclic GMP signaling pathway in an in vitro reperfusion model Br J Pharmacol 133: 841–848

    Article  PubMed  CAS  Google Scholar 

  20. Takuma K, Phuagphong P, Lee E, Mori K, Baba A, Matsuda T, 2001 Anti-apoptotic effect of c GMP in cultured astrocytes: inhibition by c GMP-dependent protein kinase of mitochondrial permeable transition pore J Biol Chem 276: 48093–48099

    PubMed  CAS  Google Scholar 

  21. Beren ME, Giblin JR, Dougherty DV, Hoifodt HK, Tveit K, Rosenblum ML, 1988 Comparison of in vitro cloning assys for drug sensitivity testing of human brain tumours Br J Neurosurg 2: 227–234

    Google Scholar 

  22. Boiardi A, Silvani A, Milanesi I, Botturi M, Broggi G, 1991 Primary glial tumor patients treated by combining cisplatin and etoposide J Neuro-Oncol 11: 165–170

    Article  CAS  Google Scholar 

  23. Morgavi P, Cimoli G, Ottoboni C, Michelotti A, Conte P, Parodi S, Russo P., 1995 Sensitization of human glioblastoma T98G cells to VP16 and VM26 by human tumor necrosis factor Anticancer Res 15: 1423–1428

    PubMed  CAS  Google Scholar 

  24. Keane MM, Ettenberg SA, Nau MM, Russell EK, Lipkowitz S, 1999 Chemotherapy augments TRAIL-induced apoptosis in breast cell lines Cancer Res 59:734–741

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neurosurgery, Zhongnan Hospital, Wuhan University, 430071, Wuhan, Hu Bei Province, People’s Republic of China

    Zhi-Min Wu, Xian-Hou Yuan, Pu-Cha Jiang, Zhi-Qiang Li & Tao Wu

  2. Department of Neurosurgery, Zhongnan Hospital, Wuhan University, 430071, Wuhan, Hu Bei Province, People’s Republic of China

    Xian-Hou Yuan

Authors
  1. Zhi-Min Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xian-Hou Yuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pu-Cha Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhi-Qiang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tao Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xian-Hou Yuan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wu, ZM., Yuan, XH., Jiang, PC. et al. Antisense oligonucleodes targeting the focal adhesion kinase inhibit proliferation, induce apoptosis and cooperate with cytotoxic drugs in human glioma cells. J Neurooncol 77, 117–123 (2006). https://doi.org/10.1007/s11060-005-9025-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11060-005-9025-9

Keywords

Search

Navigation