Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Enhanced apoptosis following treatment with TRA-8 anti-human DR5 monoclonal antibody and overexpression of exogenous Bax in human glioma cells

Gene Therapy volume 11, pages 658–667 (2004)Cite this article

Abstract

Specific activation of apoptosis in tumor cells offers a promising approach for cancer therapy. Induction of apoptosis leads to activation of specific proteases. Two major pathways for caspase activation in mammalian cells have been described. One apoptotic pathway involves members of the tumor necrosis factor family of cytokine receptors (eg death receptor 5 (DR5)). The other pathway is controlled by the Bcl-2 family of proteins. The purpose of this study was to investigate whether increased apoptosis occurs in human glioma cells following infection with a recombinant adenoviral vector encoding the human Bax gene under the control of human vascular endothelial growth factor (VEGF) promoter element (AdVEGFBax) in combination with an anti-human DR5 monoclonal antibody (TRA-8). Specific overexpression of exogenous Bax protein induced apoptosis and cell death in glioma cell lines, through activation of both caspase-8 and -9, leading to activation of downstream caspase-3. The relative sensitivity to AdVEGFBax for the glioma cell lines was U251MG>U373MG>U87MG>D54MG. The recently characterized TRA-8 monoclonal antibody induces apoptosis of most TRAIL-sensitive tumor cells by specific binding to DR5 receptors on the cellular membrane. TRA-8 induced rapid apoptosis and cell death in glioma cells, but did not demonstrate detectable cytotoxicity of primary normal human astrocytes. The efficiency of TRA-8-induced apoptosis was variable in different glioma cell lines. The relative sensitivity to TRA-8 was U373MG>U87MG>U251MG>D54MG. The combination of TRA-8 treatment and overexpression of Bax overcame TRA-8 resistance of glioma cells in vitro. Cell viability of U251MG cells was 71.1% for TRA-8 (100 ng/ml) alone, 75.9% for AdVEGFBax (5 MOI) alone and 41.1% for their combination as measured by MTS assay. Similar enhanced apoptosis results were obtained for the other glioma cell lines. In vivo studies demonstrated that the combined treatment significantly (P<0.05) suppressed the growth of U251MG xenografts and produced 60% complete tumor regressions without recurrence. These data suggest that the combination of TRA-8 treatment with specific overexpression of Bax using AdVEGFBax may be an effective approach for the treatment of human malignant gliomas.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  1. Reed JC . Mechanisms of apoptosis. Am J Pathol 2000; 157: 1415–1430.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Gross A, McDonnell J, Korsmeyer S . BCL-2 family members and the mitochondria in apoptosis. Genes Dev 1999; 13: 1899–1911.

    Article  CAS  PubMed  Google Scholar 

  3. Kagawa S et al. Antitumor effect of adenovirus-mediated Bax gene transfer on p53-sensitive and p53-resistant cancer lines. Cancer Res 2000; 60: 1157–1161.

    CAS  PubMed  Google Scholar 

  4. Xiang J et al. Pro-apoptotic treatment with an adenovirus encoding Bax enhances the effect of chemotherapy in ovarian cancer. J Gene Med 2000; 2: 97–106.

    Article  CAS  PubMed  Google Scholar 

  5. Bodmer JL et al. TRAIL receptor-2 signals apoptosis through FADD and caspase-8. Nat Cell Biol 2000; 4: 241–243.

    Article  Google Scholar 

  6. Ashkenazi A et al. Safety and antitumor activity of recombinant soluble Apo2 ligand. J Clin Invest 1999; 104: 155–162.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Walczak H et al. Tumoricidal activity of tumor necrosis factor-related apoptosis-inducing ligand in vivo. Nat Med 1999; 5: 157–163.

    Article  CAS  PubMed  Google Scholar 

  8. Ichikawa K et al. Tumoricidal activity of a novel anti-human DR5 monoclonal antibody without hepatocyte cytotoxicity. Nat Med 2001; 8: 954–960.

    Article  Google Scholar 

  9. Kaliberov S et al. Adenovirus-mediated transfer of BAX driven by the vascular endothelial growth factor promoter induces apoptosis in lung cancer cells. Mol Ther 2002; 6: 190–198.

    Article  CAS  PubMed  Google Scholar 

  10. Van Engeland M et al. A novel assay to measure loss of plasma membrane asymmetry during apoptosis of adherent cells in culture. Cytometry 1996; 24: 131–139.

    Article  CAS  PubMed  Google Scholar 

  11. Yung WK . New approaches to molecular therapy of brain tumors. Curr Opin Neurol 1994; 7: 501–505.

    Article  CAS  PubMed  Google Scholar 

  12. Shinoura N et al. Adenovirus-mediated transfer of bax with caspase-8 controlled by myelin basic protein promoter exerts an enhanced cytotoxic effect in gliomas. Cancer Gene Ther 2000; 7: 739–748.

    Article  CAS  PubMed  Google Scholar 

  13. Lowe SL et al. Prostate-specific expression of Bax delivered by an adenoviral vector induces apoptosis in LNCaP prostate cancer cells. Gene Therapy 2001; 8: 1363–1371.

    Article  CAS  PubMed  Google Scholar 

  14. Folkman J . What is the evidence that tumors are angiogenesis dependent? J Natl Cancer Inst 1990; 82: 4–6.

    Article  CAS  PubMed  Google Scholar 

  15. Arbisher JL et al. Oncogenic H-ras stimulates tumor angiogenesis by two distinct pathways. Proc Natl Acad Sci USA 1997; 94: 861–866.

    Article  Google Scholar 

  16. Grugel S et al. Both v-Ha-Ras and v-Raf stimulate expression of the vascular endothelial growth factor in NIH 3T3 cells. J Biol Chem 1995; 270: 25915–25919.

    Article  CAS  PubMed  Google Scholar 

  17. Mukhopadhyay D, Tsiokas L, Sukhatme VP . Wild-type p53 and v-Src exert opposing influences on human vascular endothelial growth factor gene expression. Cancer Res 1995; 55: 6161–6165.

    CAS  PubMed  Google Scholar 

  18. Kieser A et al. Mutant p53 potentiates protein kinase C induction of vascular endothelial growth factor expression. Oncogene 1994; 9: 963–969.

    CAS  PubMed  Google Scholar 

  19. Fukumura D et al. Hypoxia and acidosis independently up-regulate vascular endothelial growth factor transcription in brain tumors in vivo. Cancer Res 2001; 61: 6020–6024.

    CAS  PubMed  Google Scholar 

  20. Plate KH, Risau W . Angiogenesis in malignant gliomas. Glia 1995; 15: 339–347.

    Article  CAS  PubMed  Google Scholar 

  21. Takekawa Y, Sawada T . Vascular endothelial growth factor and neovascularization in astrocytic tumors. Pathol Int 1998; 48: 109–114.

    Article  CAS  PubMed  Google Scholar 

  22. Park WS et al. Inactivating mutations of KILLER/DR5 gene in gastric cancers. Gastroenterology 2001; 121: 219–225.

    Article  Google Scholar 

  23. Irmler M et al. Inhibition of death receptor signals by cellular FLIP. Nature 1997; 388: 190–195.

    Article  CAS  PubMed  Google Scholar 

  24. Hopkins-Donaldson S et al. Loss of caspase-8 expression in neuroblastoma is related to malignancy and resistance to TRAIL-induced apoptosis. Med Pediatr Oncol 2000; 35: 608–611.

    Article  CAS  PubMed  Google Scholar 

  25. Eggert A et al. Resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in neuroblastoma cells correlates with a loss of caspase-8 expression. Cancer Res 2001; 61: 1314–1319.

    CAS  PubMed  Google Scholar 

  26. Burns TF, El-Deiry WS . Identification of inhibitors of TRAIL-induced death (ITIDs) in the TRAIL-sensitive colon carcinoma cell line SW480 using a genetic approach. J Biol Chem 2001; 276: 37879–37886.

    CAS  PubMed  Google Scholar 

  27. Lamothe B, Aggarwal BB . Ectopic expression of Bcl-2 and Bcl-xL inhibits apoptosis induced by TNF-related apoptosis-inducing ligand (TRAIL) through suppression of caspases-8, 7, and 3 and BID cleavage in human acute myelogenous leukemia cell line HL-60. J Interferon Cytokine Res 2002; 22: 269–279.

    Article  CAS  PubMed  Google Scholar 

  28. Ravi R, Bedi A . Requirement of BAX for TRAIL/Apo2L-induced apoptosis of colorectal cancers: synergism with sulindac-mediated inhibition of Bcl-x(L). Cancer Res 2002; 62: 1583–1587.

    CAS  PubMed  Google Scholar 

  29. Luo X et al. Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell 1998; 94: 481–490.

    Article  CAS  PubMed  Google Scholar 

  30. Yin XM . Signal transduction mediated by Bid, a pro-death Bcl-2 family proteins, connects the death receptor and mitochondria apoptosis pathways. Cell Res 2000; 10: 161–167.

    Article  CAS  PubMed  Google Scholar 

  31. LeBlanc H et al. Tumor-cell resistance to death receptor–induced apoptosis through mutational inactivation of the proapoptotic Bcl-2 homolog Bax. Nat Med 2002; 8: 274–281.

    Article  CAS  PubMed  Google Scholar 

  32. Deng Y, Lin Y, Wu X . TRAIL-induced apoptosis requires Bax-dependent mitochondrial release of Smac/DIABLO. Genes Dev 2002; 16: 33–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Peter ME et al. The death receptors. Results Probl Cell Differ 1999; 23: 25–63.

    Article  CAS  PubMed  Google Scholar 

  34. Sprick MR et al. FADD/MORT1 and caspase-8 are recruited to TRAIL receptors 1 and 2 and are essential for apoptosis mediated by TRAIL receptor 2. Immunity 2000; 12: 599–609.

    Article  CAS  PubMed  Google Scholar 

  35. Zhang H et al. BAR: an apoptosis regulator at the intersection of caspases and Bcl-2 family proteins. Proc Natl Acad Sci USA 2000; 97: 2597–2602.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Stegh AH et al. Inactivation of caspase-8 on mitochondria of Bcl-xL-expressing MCF7-Fas cells: role for the bifunctional apoptosis regulator protein. J Biol Chem 2002; 277: 4351–4360.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Radiation Oncology, University of Alabama at Birmingham, Birmingham, AL, USA

    S Kaliberov, M A Stackhouse & D J Buchsbaum

  2. Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA

    L Kaliberova & T Zhou

Authors
  1. S Kaliberov
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M A Stackhouse
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L Kaliberova
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. D J Buchsbaum
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supported by Sankyo Co., Ltd and Pediatric Brain Tumor Foundation of the United States Grant #000136268

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kaliberov, S., Stackhouse, M., Kaliberova, L. et al. Enhanced apoptosis following treatment with TRA-8 anti-human DR5 monoclonal antibody and overexpression of exogenous Bax in human glioma cells. Gene Ther 11, 658–667 (2004). https://doi.org/10.1038/sj.gt.3302215

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3302215

Keywords

This article is cited by

Search

Advanced search

Quick links