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. 2006 Apr 5;12(1):12–20. doi: 10.1111/j.1750-3639.2002.tb00418.x

CD95‐mediated Apoptosis of Human Glioma Cells: Modulation by Epidermal Growth Factor Receptor Activity

Joachim P Steinbach 1,✉, Petra Supra 1, H‐J Su Huang 2, Webster K Cavenee 2, Michael Weller 1
PMCID: PMC8095827  PMID: 11770895

Abstract

The death ligands CD95L and Apo2L/TRAIL are promising investigational agents for the treatment of malignant glioma. EGFR is overexpressed in a significant proportion of malignant gliomas in vivo. Here, we report that CD95L‐induced cell death is enhanced by EGFR inhibition using tyrphostine AG1478 in 7 of 12 human malignant glioma cell lines. Conversely, CD95‐mediated and Apo2L‐induced cell death are both inhibited by overexpression of EGFR in LN‐229 cells. CD95L‐induced cell death augmented by AG1478 is accompanied by enhanced processing of caspase 8. LN‐229 cells overexpressing the viral caspase inhibitor, crm‐A, are not sensitized to CD95L‐induced cell death by AG1478, indicating that EGFR exerts its antiapoptotic properties through a caspase 8‐dependent pathway. These data define a modulatory effect of EGFR‐activity on death ligand‐induced apoptosis and indicate that EGFR inhibition is likely to improve the efficacy of death ligand‐based cancer therapies. Furthermore, it is tempting to speculate that EGFR amplification protects tumor cells from death ligand‐mediated host immune responses in vivo and that EGFR's effects on death receptor‐mediated apoptosis may explain the anti‐tumor effects of non‐cytotoxic, unarmed anti‐EGFR family antibodies.

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References

  • 1. Aboud‐Pirak E, Hurwitz E, Pirak ME, Bellot F, Schlessinger J, Sela M (1988) Efficacy of antibodies to epidermal growth factor receptor against KB carcinoma in vitro and in nude mice. J Natl Cancer Inst 80:1605–1611. [DOI] [PubMed] [Google Scholar]
  • 2. Ashkenazi A, Dixit VM (1999) Apoptosis control by death and decoy receptors. Curr Opin Cell Biol 11:255–260. [DOI] [PubMed] [Google Scholar]
  • 3. Bigner SH, Humphrey PA, Wong AJ, Vogelstein B, Mark J, Friedman HS, Bigner DD (1990) Characterization of the epidermal growth factor receptor in human glioma cell lines and xenografts. Cancer Res 50:8017–8022. [PubMed] [Google Scholar]
  • 4. Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS, Anderson MJ, Arden KC, Blenis J, Greenberg ME (1999) Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 96:857–868. [DOI] [PubMed] [Google Scholar]
  • 5. Ciardiello F, Caputo R, Troiani T, Borriello G, Kandimalla ER, Agrawal S, Mendelsohn J, Bianco AR, Tortora G (2001) Antisense oligonucleotides targeting the epidermal growth factor receptor inhibit proliferation, induce apoptosis, and cooperate with cytotoxic drugs in human cancer cell lines. Int J Cancer 93:172–178. [DOI] [PubMed] [Google Scholar]
  • 6. Cobleigh MA, Vogel CL, Tripathy D, Robert NJ, Scholl S, Fehrenbacher L, Wolter JM, Paton V, Shak S, Lieberman G, Slamon DJ (1999) Multinational study of the efficacy and safety of humanized anti‐HER2 monoclonal antibody in women who have HER2‐overexpressing metastatic breast cancer that has progressed after chemotherapy for metastatic disease. J Clin Oncol 17:2639–2648. [DOI] [PubMed] [Google Scholar]
  • 7. Dixit M, Yang JL, Poirier MC, Price JO, Andrews PA, Arteaga CL (1997) Abrogation of cisplatin‐induced programmed cell death in human breast cancer cells by epidermal growth factor antisense RNA. J Natl Cancer Inst 89:365–373. [DOI] [PubMed] [Google Scholar]
  • 8. Fry DW, Kraker AJ, McMichael A, Ambroso LA, Nelson JM, Leopold WR, Connors RW, Bridges AJ (1994) A specific inhibitor of the epidermal growth factor receptor tyrosine kinase. Science 265:1093–1095. [DOI] [PubMed] [Google Scholar]
  • 9. Furnari FB, Lin H, Huang HS, Cavenee WK (1997) Growth suppression of glioma cells by PTEN requires a functional phosphatase catalytic domain. Proc Natl Acad Sci U S A 94:12479–12484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10. Gibson S, Tu S, Oyer R, Anerson SM, Johnson GL (1999) Epidermal growth factor protects epithelial cells against Fas‐induced apoptosis. J Biol Chem 274:17612–17618. [DOI] [PubMed] [Google Scholar]
  • 11. Glaser T, Wagenknecht B, Weller M (2001) Identification of p21 as a target of cycloheximide‐mediated facilitation of CD95‐mediated apoptosis in human malignant glioma cells. Oncogene In press. [DOI] [PubMed] [Google Scholar]
  • 12. Han Y, Caday CG, Nanda A, Cavenee WK, Huang HJ (1996) Tyrphostin AG 1478 preferentially inhibits human glioma cells expressing truncated rather than wild‐type epidermal growth factor receptors. Cancer Res 56:3859–3861. [PubMed] [Google Scholar]
  • 13. Holland EC, Celestino J, Dai C, Schaefer L, Sawaya RE, Fuller GN (2000) Combined activation of Ras and Akt in neural progenitors induces glioblastoma formation in mice. Nat Genet 25:55–57. [DOI] [PubMed] [Google Scholar]
  • 14. Levitzki A, Gazit A (1995) Tyrosine kinase inhibition: an approach to drug development. Science 267:1782–1788. [DOI] [PubMed] [Google Scholar]
  • 15. Nagane M, Levitzki A, Gazit A, Cavenee WK, Huang HJ (1998) Drug resistance of human glioblastoma cells conferred by a tumor‐ specific mutant epidermal growth factor receptor through modulation of Bcl‐XL and caspase‐3‐like proteases. Proc Natl Acad Sci U S A 95:5724–5729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16. Nagane M, Pan G, Weddle JJ, Dixit VM, Cavenee WK, Huang HJ (2000) Increased death receptor 5 expression by chemotherapeutic agents in human gliomas causes synergistic cytotoxicity with tumor necrosis factor‐related apoptosis‐inducing ligand in vitro and in vivo. Cancer Res 60:847–853. [PubMed] [Google Scholar]
  • 17. Nishikawa R, Ji XD, Harmon RC, Lazar CS, Gill GN, Cavenee WK, Huang HJ (1994) A mutant epidermal growth factor receptor common in human glioma confers enhanced tumorigenicity. Cancer Res 91:7727–7731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18. O'Rourke DM, Qian X, Zhang HT, Davis JG, Nute E, Meinkoth J, Greene MI (1997) Trans receptor inhibition of human glioblastoma cells by erbB family ectodomains. Proc Natl Acad Sci U S A 94:3250–3255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. Panka DJ, Mano T, Suhara T, Walsh K, Mier JW (2001) Phosphatidylinositol‐3 Kinase/Akt activity regulates c‐FLIP expression in tumor cells. J Biol Chem 276:6893–6896. [DOI] [PubMed] [Google Scholar]
  • 20. Pegram M, Slamon D (2000) Biological rationale for HER2/neu (c‐erbB2) as a target for monoclonal antibody therapy. Semin Oncol 27:13–19. [PubMed] [Google Scholar]
  • 21. Peli J, Schroter M, Rudaz C, Hahne M, Meyer C, Reichmann E, Tschopp J (1999) Oncogenic Ras inhibits Fas ligand‐mediated apoptosis by downregulating the expression of Fas. Embo J 18:1824–1831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22. Redemann N, Holzmann B, von Ruden T, Wagner EF, Schlessinger J, Ullrich A (1992) Anti‐oncogenic activity of signalling‐defective epidermal growth factor receptor mutants. Mol Cell Biol 12:491–498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. Rieger J, Naumann U, Glaser T, Ashkenazi A, Weller M (1998) APO2 ligand: a novel lethal weapon against malignant glioma FEBS Lett 427:124–128. [DOI] [PubMed] [Google Scholar]
  • 24. Roth W, Fontana A, Trepel M, Reed JC, Dichgans J, Weller M (1997) Immunochemotherapy of malignant glioma: synergistic activity of CD95 ligand and chemotherapeutics. Cancer Immunol Immunother 44:55–63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. Roth W, Isenmann S, Naumann U, Kugler S, Bahr M, Dichgans J, Ashkenazi A, Weller M (1999) Locoregional Apo2L/TRAIL eradicates intracranial human malignant glioma xenografts in athymic mice in the absence of neurotoxicity. Biochem Biophys Res Commun 265:479–483. [DOI] [PubMed] [Google Scholar]
  • 26. Sampson JH, Crotty LE, Lee S, Archer GE, Ashley DM, Wikstrand CJ, Hale LP, Small C, Dranoff G, Friedman AH, Friedman HS, Bigner DD (2000) Unarmed, tumor‐specific monoclonal antibody effectively treats brain tumors. Proc Natl Acad Sci U S A 97:7503–7508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27. Schmidt F, Rieger J, Wischhusen J, Naumann U, Weller M (2001) Glioma cell sensitivity to topotecan: the role of p53 and topotecan‐induced DNAdamage. Eur J Pharmacol 412:21–25. [DOI] [PubMed] [Google Scholar]
  • 28. Takeda K, Hayakawa Y, Smyth MJ, Kayagaki N, Yamaguchi N, Kakuta S, Iwakura Y, Yagita H, Okumura K (2001) Involvement of tumor necrosis factor‐related apoptosis‐inducing ligand in surveillance of tumor metastasis by liver natural killer cells. Nat Med 7:94–100. [DOI] [PubMed] [Google Scholar]
  • 29. Vincent PW, Bridges AJ, Dykes DJ, Fry DW, Leopold WR, Patmore SJ, Roberts BJ, Rose S, Sherwood V, Zhou H, Elliott WL (2000) Anticancer efficacy of the irreversible EGFR tyrosine kinase inhibitor PD 0169414 against human tumor xenografts. Cancer Chemother Pharmacol 45:231–238. [DOI] [PubMed] [Google Scholar]
  • 30. Wagenknecht B, Glaser T, Naumann U, Kugler S, Isenmann S, Bahr M, Ksorneluk R, Liston P, Weller M (1999) Expression and biological activity of X‐linked inhibitor of apoptosis (XIAP) in human malignant glioma. Cell Death Differ 6:370–376. [DOI] [PubMed] [Google Scholar]
  • 31. Wagenknecht B, Schulz JB, Gulbins E, Weller M (1998) Crm‐A, bcl‐2 and NDGA inhibit CD95L‐induced apoptosis of malignant glioma cells at the level of caspase 8 processing. Cell Death Differ 5:894–900. [DOI] [PubMed] [Google Scholar]
  • 32. Watanabe K, Tachibana O, Sata K, Yonekawa Y, Kleihues P, Ohgaki H (1996) Overexpression of the EGF receptor and p53 mutations are mutually exclusive in the evolution of primary and secondary glioblastomas. Brain Pathol 6:217–223. [DOI] [PubMed] [Google Scholar]
  • 33. Weber RG, Rieger J, Naumann U, Lichter P, Weller M (2001) Chromosomal imbalances associated with response to chemotherapy and cytotoxic cytokines in human malignant glioma cell lines. Int J Cancer 91:213–218. [DOI] [PubMed] [Google Scholar]
  • 34. Weller M, Frei K, Groscurth P, Krammer PH, Yonekawa Y, Fontana A (1994) Anti‐Fas/APO‐1 antibody‐mediated apoptosis of cultured human glioma cells. Induction and modulation of sensitivity by cytokines. J Clin Invest 94:954–964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35. Weller M, Malipiero U, Aguzzi A, Reed JC, Fontana A (1995) Protooncogene bcl‐2 gene transfer abrogates Fas/APO‐1 antibody‐mediated apoptosis of human malignant glioma cells and confers resistance to chemotherapeutic drugs and therapeutic irradiation. J Clin Invest 95:2633–2643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36. Weller M, Rieger J, Grimmel C, Van Meir EG, De Tribolet N, Krajewski S, Reed JC, von Deimling A, Dichgans J (1998) Predicting chemoresistance in human malignant glioma cells: the role of molecular genetic analyses. Int J Cancer 79:640–644. [DOI] [PubMed] [Google Scholar]
  • 37. Wick W, Furnari FB, Naumann U, Cavenee WK, Weller M (1999) PTEN gene transfer in human malignant glioma: sensitization to irradiation and CD95L‐induced apoptosis. Oncogene 18:3936–3943. [DOI] [PubMed] [Google Scholar]
  • 38. Winter S, Roth W, Dichgans J, Weller M (1998) Synergistic activity of CD95 ligand and teniposide (VM26) does not involve enhanced formation of cleavable DNA complexes or changes in CD95/CD95 ligand expression. Eur J Pharmacol 341:323–328. [DOI] [PubMed] [Google Scholar]

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