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.

  • Original Article
  • Published:

Death receptor 4 (DR4) efficiently kills breast cancer cells irrespective of their sensitivity to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)

Cancer Gene Therapy volume 11, pages 691–698 (2004)Cite this article

Abstract

Breast cancer cells are generally resistant to induction of apoptosis by treatment with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). In this study, we demonstrate that both TRAIL-sensitive and TRAIL-resistant breast cancer cell lines can be efficiently killed by overexpression of the TRAIL receptor, death receptor 4 (DR4). The extent of cell death depended on the strength of the promoter driving DR4 expression. When driven by the strong CMV promoter, expression of DR4 killed over 90% of cells in five out of six cell lines tested in the absence of exogenous TRAIL. When driven by the relatively weak tumor-specific hTERT promoter, DR4 was less effective alone, but sensitized cells to killing by TRAIL. The extent of TRAIL sensitization depended on the magnitude of hTERT promoter activity. MCF-7 cells were relatively resistant to the action of DR4. We compared expression of the genes involved in transduction and execution of the death receptor-initiated apoptotic stimuli between MCF-7 and DR4-sensitive cell lines. We confirmed that in the panel of cell lines, MCF-7 was the only line deficient in expression of caspase 3. Bcl-2 and FLIP proteins, implicated in suppression of TRAIL-induced apoptosis, were expressed at a higher level.

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

Similar content being viewed by others

References

  1. Held J, Schulze-Osthoff K . Potential and caveats of TRAIL in cancer therapy. Drug Resist Update. 2001;4:243–252.

    Article  CAS  Google Scholar 

  2. Nagane M, Huang HJ, Cavenee WK . The potential of TRAIL for cancer chemotherapy. Apoptosis. 2001;6:191–197.

    Article  CAS  PubMed  Google Scholar 

  3. Van Ophoven A, Ng CP, Patel B, Bonavida B, Belldegrun A . Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) for treatment of prostate cancer: first results and review of the literature. Prostate Cancer Prostatic Dis. 1999;2:227–233.

    Article  CAS  PubMed  Google Scholar 

  4. Keane MM, Rubinstein Y, Cuello M, et al. Inhibition of NF-kappaB activity enhances TRAIL mediated apoptosis in breast cancer cell lines. Breast Cancer Res Treat. 2000;64:211–219.

    Article  CAS  PubMed  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  6. Ashkenazi A, Dixit VM . Apoptosis control by death and decoy receptors. Curr Opin Cell Biol. 1999;11:255–260.

    Article  CAS  PubMed  Google Scholar 

  7. Bernard D, Quatannens B, Vandenbunder B, Abbadie C . Rel/NF-kappaB transcription factors protect against tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-induced apoptosis by up-regulating the TRAIL decoy receptor DcR1. J Biol Chem. 2001;276:27322–27328.

    Article  CAS  PubMed  Google Scholar 

  8. LeBlanc HN, Ashkenazi A . Apo2L/TRAIL and its death and decoy receptors. Cell Death Differ. 2003;10:66–75.

    Article  CAS  PubMed  Google Scholar 

  9. Roth W, Isenmann S, Nakamura M, et al. Soluble decoy receptor 3 is expressed by malignant gliomas and suppresses CD95 ligand-induced apoptosis and chemotaxis. Cancer Res. 2001;61:2759–2765.

    CAS  PubMed  Google Scholar 

  10. Seitz S, Wassmuth P, Fischer J, et al. Mutation analysis and mRNA expression of trail-receptors in human breast cancer. Int J Cancer. 2002;102:117–128.

    Article  CAS  PubMed  Google Scholar 

  11. Shin MS, Kim HS, Lee SH, et al. Mutations of tumor necrosis factor-related apoptosis-inducing ligand receptor 1 (TRAIL-R1) and receptor 2 (TRAIL-R2) genes in metastatic breast cancers. Cancer Res. 2001;61:4942–4946.

    CAS  PubMed  Google Scholar 

  12. Hopkins-Donaldson S, Ziegler A, Kurtz S, et al. Silencing of death receptor and caspase-8 expression in small cell lung carcinoma cell lines and tumors by DNA methylation. Cell Death Differ. 2003;10:356–364.

    Article  CAS  PubMed  Google Scholar 

  13. Ozoren N, Fisher MJ, Kim K, et al. Homozygous deletion of the death receptor DR4 gene in a nasopharyngeal cancer cell line is associated with TRAIL resistance. Int J Oncol. 2000;16:917–925.

    CAS  PubMed  Google Scholar 

  14. Park WS, Lee JH, Shin MS, et al. Inactivating mutations of KILLER/DR5 gene in gastric cancers. Gastroenterology. 2001;121:1219–1225.

    Article  CAS  PubMed  Google Scholar 

  15. Bin L, Li X, Xu LG, Shu HB . The short splice form of Casper/c-FLIP is a major cellular inhibitor of TRAIL-induced apoptosis. FEBS Lett. 2002;510:37–40.

    Article  CAS  PubMed  Google Scholar 

  16. Bullani RR, Huard B, Viard-Leveugle I, et al. Selective expression of FLIP in malignant melanocytic skin lesions. J Invest Dermatol. 2001;117:360–364.

    Article  CAS  PubMed  Google Scholar 

  17. Kelly MM, Hoel BD, Voelkel-Johnson C . Doxorubicin pretreatment sensitizes prostate cancer cell lines to TRAIL induced apoptosis which correlates with the loss of c-FLIP expression. Cancer Biol Ther. 2002;1:520–527.

    Article  PubMed  Google Scholar 

  18. Kim Y, Suh N, Sporn M, Reed JC . An inducible pathway for degradation of FLIP protein sensitizes tumor cells to TRAIL-induced apoptosis. J Biol Chem. 2002;277:22320–22329.

    Article  CAS  PubMed  Google Scholar 

  19. Kim IK, Jung YK, Noh DY, et al. Functional screening of genes suppressing TRAIL-induced apoptosis: distinct inhibitory activities of Bcl-XL and Bcl-2. Br J Cancer. 2003;88:910–917.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Fulda S, Meyer E, Debatin KM . Inhibition of TRAIL-induced apoptosis by Bcl-2 overexpression. Oncogene. 2002;21:2283–2294.

    Article  CAS  PubMed  Google Scholar 

  21. 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 

  22. Ng CP, Bonavida B . X-linked inhibitor of apoptosis (XIAP) blocks Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis of prostate cancer cells in the presence of mitochondrial activation: sensitization by overexpression of second mitochondria-derived activator of caspase/direct IAP-binding protein with low pl (Smac/DIABLO). Mol Cancer Ther. 2002;1:1051–1058.

    CAS  PubMed  Google Scholar 

  23. Nesterov A, Lu X, Johnson M, et al. Elevated AKT activity protects the prostate cancer cell line LNCaP from TRAIL-induced apoptosis. J Biol Chem. 2001;276:10767–10774.

    Article  CAS  PubMed  Google Scholar 

  24. Panka DJ, Mano T, Suhara T, Walsh K, Mier JW . Phosphatidylinositol 3-kinase/Akt activity regulates c-FLIP expression in tumor cells. J Biol Chem. 2001;276:6893–6896.

    Article  CAS  PubMed  Google Scholar 

  25. Mitsiades CS, Mitsiades N, Poulaki V, et al. Activation of NF-kappaB and upregulation of intracellular anti-apoptotic proteins via the IGF-1/Akt signaling in human multiple myeloma cells: therapeutic implications. Oncogene. 2002;21:5673–5683.

    Article  CAS  PubMed  Google Scholar 

  26. Secchiero P, Gonelli A, Carnevale E, et al. TRAIL promotes the survival and proliferation of primary human vascular endothelial cells by activating the Akt and ERK pathways. Circulation. 2003;107:2250–2256.

    Article  PubMed  Google Scholar 

  27. Degli-Esposti MA, Dougall WC, Smolak PJ, et al. The novel receptor TRAIL-R4 induces NF-kappaB and protects against TRAIL-mediated apoptosis, yet retains an incomplete death domain. Immunity. 1997;7:813–820.

    Article  CAS  PubMed  Google Scholar 

  28. Blanc C, Deveraux QL, Krajewski S, et al. Caspase-3 is essential for procaspase-9 processing and cisplatin-induced apoptosis of MCF-7 breast cancer cells. Cancer Res. 2000;60:4386–4390.

    CAS  PubMed  Google Scholar 

  29. Devarajan E, Sahin AA, Chen JS, et al. Down-regulation of caspase 3 in breast cancer: a possible mechanism for chemoresistance. Oncogene. 2002;21:8843–8851.

    Article  CAS  PubMed  Google Scholar 

  30. Papoff G, Hausler P, Eramo A, et al. Identification and characterization of a ligand-independent oligomerization domain in the extracellular region of the CD95 death receptor. J Biol Chem. 1999;274:38241–38250.

    Article  CAS  PubMed  Google Scholar 

  31. Faubion WA, Guicciardi ME, Miyoshi H, et al. Toxic bile salts induce rodent hepatocyte apoptosis via direct activation of Fas. J Clin Invest. 1999;103:137–145.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Gu J, Kagawa S, Takakura M, et al. Tumor-specific transgene expression from the human telomerase reverse transcriptase promoter enables targeting of the therapeutic effects of the Bax gene to cancers. Cancer Res. 2000;60:5359–5364.

    CAS  PubMed  Google Scholar 

  33. Koga S, Hirohata S, Kondo Y, et al. A novel telomerase-specific gene therapy: gene transfer of caspase-8 utilizing the human telomerase catalytic subunit gene promoter. Hum Gene Ther. 2000;11:1397–1406.

    Article  CAS  PubMed  Google Scholar 

  34. Komata T, Kondo Y, Kanzawa T, et al. Caspase-8 gene therapy using the human telomerase reverse transcriptase promoter for malignant glioma cells. Hum Gene Ther. 2002;13:1015–1025.

    Article  CAS  PubMed  Google Scholar 

  35. Lin T, Huang X, Gu J, et al. Long-term tumor-free survival from treatment with the GFP-TRAIL fusion gene expressed from the hTERT promoter in breast cancer cells. Oncogene. 2002;21:8020–8028.

    Article  CAS  PubMed  Google Scholar 

  36. Hietakangas V, Poukkula M, Heiskanen KM, et al. Erythroid differentiation sensitizes K562 leukemia cells to TRAIL-induced apoptosis by downregulation of c-FLIP. Mol Cell Biol. 2003;23:1278–1291.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Xiao C, Yang BF, Asadi N, Beguinot F, Hao C . Tumor necrosis factor-related apoptosis-inducing ligand-induced death-inducing signaling complex and its modulation by c-FLIP and PED/PEA-15 in glioma cells. J Biol Chem. 2002;277:25020–25025.

    Article  CAS  PubMed  Google Scholar 

  38. Xue LY, Chiu SM, Oleinick NL . Staurosporine-induced death of MCF-7 human breast cancer cells: a distinction between caspase-3-dependent steps of apoptosis and the critical lethal lesions. Exp Cell Res. 2003;283:135–145.

    Article  CAS  PubMed  Google Scholar 

  39. Xue LY, Chiu SM, Oleinick NL . Photodynamic therapy-induced death of MCF-7 human breast cancer cells: a role for caspase-3 in the late steps of apoptosis but not for the critical lethal event. Exp Cell Res. 2001;263:145–155.

    Article  CAS  PubMed  Google Scholar 

  40. Suyama E, Kawasaki H, Taira K . Identification of a caspase 3-independent role of pro-apoptotic factor Bak in TNF-alpha-induced apoptosis. FEBS Lett. 2002;528:63–69.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr Satoru Kyo for kindly supplying us with the hTERT promoter constructs. This work was supported by the NIH 5 K12 CA01723-10 Physician Scientist Training Grant and an Institutional Research Grant from American Cancer Society.

Author information

Authors and Affiliations

  1. Department of Medicine, Division of Medical Oncology, University of Texas Health Science Center at San Antonio, Texas, USA

    Irene Kazhdan & Robert A Marciniak

  2. South Texas Veterans Health Care System, Texas, USA

    Robert A Marciniak

Authors
  1. Irene Kazhdan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Robert A Marciniak
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Irene Kazhdan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kazhdan, I., Marciniak, R. Death receptor 4 (DR4) efficiently kills breast cancer cells irrespective of their sensitivity to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Cancer Gene Ther 11, 691–698 (2004). https://doi.org/10.1038/sj.cgt.7700747

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.cgt.7700747

Keywords

This article is cited by

Search

Advanced search

Quick links