Skip to main content
SpringerLink
Log in

Inhibition of novel protein kinase Cɛ augments TRAIL-induced cell death in A549 lung cancer cells

  • Article
  • Published:
Pathology & Oncology Research

Abstract

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has great potential for cancer treatment since it provokes cell death in most tumor cells while leaving most normal cells unscathed. Some cancers, however, show resistance to TRAIL, indicating that TRAIL alone may be insufficient for cancer therapy. Here we studied whether the apoptotic susceptibility of A549 non-small cell lung cancer cells could be modulated by inhibiting protein kinase C (PKC). We show that an inhibitor with preference for novel PKC isozymes, NPC 15437, significantly augmented TRAIL sensitivity of A549 cells, as judged by assessing cell death and mitochondrial membrane potential. Likewise, NPC 15437 also significantly potentiated the responsiveness of DAOY medulloblastoma cells to TRAIL. In contrast, an inhibitor with preference for conventional PKC isozymes, Gö6976, did not augment TRAIL sensitivity of A549 cells. To further specify the PKC isozyme responsible for TRAIL sensitization, we used a peptide inhibitor with selectivity for the novel PKC isozyme ɛ, myr-PKCɛ V1-2. The inhibition of PKCɛ resulted in a significant amplification of the cytotoxic activity of TRAIL in A549 cells. Altogether, our study provides evidence for a considerable role of PKCɛ in the apoptotic responsiveness of A549 lung cancer cells, and possibly other malignancies, to TRAIL.

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. Baader E, Toloczko A, Fuchs U, et al: Tumor necrosis factor-related apoptosis-inducing ligand-mediated proliferation of tumor cells with receptor-proximal apoptosis defects. Cancer Res 65: 7888–7895, 2005.

    PubMed  CAS  Google Scholar 

  2. Basu A, Lu D, Sun B, et al: Proteolytic activation of protein kinase C-epsilon by caspase-mediated processing and transduction of antiapoptotic signals. J Biol Chem 277: 41850–41856, 2002.

    Article  PubMed  CAS  Google Scholar 

  3. Basu A, Weixel KM: Comparison of protein kinase C activity and isoform expression in cisplatin-sensitive and -resistant ovarian carcinoma cells. Int J Cancer 62: 457–460, 1995.

    Article  PubMed  CAS  Google Scholar 

  4. Carney DN: Lung cancer—time to move on from chemotherapy. N Engl J Med 346: 126–128, 2002.

    Article  PubMed  Google Scholar 

  5. Courage C, Budworth J, Gescher A: Comparison of ability of protein kinase C inhibitors to arrest cell growth and to alter cellular protein kinase C localisation. Br J Cancer 71: 697–704, 1995.

    PubMed  CAS  Google Scholar 

  6. Ding L, Wang H, Lang W, et al: Protein kinase C-epsilon promotes survival of lung cancer cells by suppressing apoptosis through dysregulation of the mitochondrial caspase pathway. J Biol Chem 277: 35305–35313, 2002.

    Article  PubMed  CAS  Google Scholar 

  7. Fan QL, Zou WY, Song LH, et al: Synergistic antitumor activity of TRAIL combined with chemotherapeutic agents in A549 cell lines in vitro and in vivo. Cancer Chemother Pharmacol 55: 189–196, 2005.

    Article  PubMed  CAS  Google Scholar 

  8. Fesik SW: Promoting apoptosis as a strategy for cancer drug discovery. Nat Rev Cancer 5: 876–885, 2005.

    Article  PubMed  CAS  Google Scholar 

  9. Frese S, Brunner T, Gugger M, et al: Enhancement of Apo2L/TRAIL (tumor necrosis factor-related apoptosis-inducing ligand)-induced apoptosis in non-small cell lung cancer cell lines by chemotherapeutic agents without correlation to the expression level of cellular protease caspase-8 inhibitory protein. J Thorac Cardiovasc Surg 123: 168–174, 2002.

    Article  PubMed  CAS  Google Scholar 

  10. Fulda S, Debatin KM: Exploiting death receptor signaling pathways for tumor therapy. Biochim Biophys Acta 1705: 27–41, 2004.

    PubMed  CAS  Google Scholar 

  11. Ganten TM, Koschny R, Sykora J, et al: Preclinical differentiation between apparently safe and potentially hepatotoxic applications of TRAIL either alone or in combination with chemotherapeutic drugs. Clin Cancer Res 12: 2640–2646, 2006.

    Article  PubMed  CAS  Google Scholar 

  12. Gillespie S, Zhang XD, Hersey P: Variable expression of protein kinase C epsilon in human melanoma cells regulates sensitivity to TRAIL-induced apoptosis. Mol Cancer Ther 4: 668–676, 2005.

    Article  PubMed  CAS  Google Scholar 

  13. Harper N, Hughes MA, Farrow SN, et al: Protein kinase C modulates TRAIL-induced apoptosis by targeting the apical events of death receptor signaling. J Biol Chem 278: 44338–44347, 2003.

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  15. Hofmann J: Protein kinase C isozymes as potential targets for anticancer therapy. Curr Cancer Drug Targets 4: 125–146, 2004.

    Article  PubMed  CAS  Google Scholar 

  16. Jemal A, Siegel R, Ward E, et al: Cancer statistics, 2006. CA Cancer J Clin 56: 106–130, 2006.

    Article  PubMed  Google Scholar 

  17. Jin H, Yang R, Fong S, et al: Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand cooperates with chemotherapy to inhibit orthotopic lung tumor growth and improve survival. Cancer Res 64: 4900–4905, 2004.

    Article  PubMed  CAS  Google Scholar 

  18. Johnson JA, Gray MO, Chen CH, et al: A protein kinase C translocation inhibitor as an isozyme-selective antagonist of cardiac function. J Biol Chem 271: 24962–24966, 1996.

    Article  PubMed  CAS  Google Scholar 

  19. Johnstone RW, Ruefli AA, Lowe SW: Apoptosis: a link between cancer genetics and chemotherapy. Cell 108: 153–164, 2002.

    Article  PubMed  CAS  Google Scholar 

  20. Kim CY, Giaccia AJ, Strulovici B, et al: Differential expression of protein kinase C epsilon protein in lung cancer cell lines by ionising radiation. Br J Cancer 66: 844–849, 1992.

    PubMed  CAS  Google Scholar 

  21. Laack E, Dickgreber N, Muller T, et al: Randomized phase III study of gemcitabine and vinorelbine versus gemcitabine, vinorelbine, and cisplatin in the treatment of advanced non-small-cell lung cancer: from the German and Swiss Lung Cancer Study Group. J Clin Oncol 22: 2348–2356, 2004.

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  23. Manna SK, Aggarwal BB: All-trans-retinoic acid upregulates TNF receptors and potentiates TNF-induced activation of nuclear factors-kappaB, activated protein-1 and apoptosis in human lung cancer cells. Oncogene 19: 2110–2119, 2000.

    Article  PubMed  CAS  Google Scholar 

  24. Martiny-Baron G, Kazanietz MG, Mischak H, et al: Selective inhibition of protein kinase C isozymes by the indolocarbazole Go 6976. J Biol Chem 268: 9194–9197, 1993.

    PubMed  CAS  Google Scholar 

  25. McJilton MA, Van Sikes C, Wescott GG, et al: Protein kinase Cepsilon interacts with Bax and promotes survival of human prostate cancer cells. Oncogene 22: 7958–7968, 2003.

    Article  PubMed  CAS  Google Scholar 

  26. Mitsiades N, Mitsiades CS, Poulaki V, et al: Intracellular regulation of tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in human multiple myeloma cells. Blood 99: 2162–2171, 2002.

    Article  PubMed  CAS  Google Scholar 

  27. Monick M, Staber J, Thomas K, et al: Respiratory syncytial virus infection results in activation of multiple protein kinase C isoforms leading to activation of mitogen-activated protein kinase. J Immunol 166: 2681–2687, 2001.

    PubMed  CAS  Google Scholar 

  28. Odoux C, Albers A, Amoscato AA, et al: TRAIL, FasL and a blocking anti-DR5 antibody augment paclitaxel-induced apoptosis in human non-small-cell lung cancer. Int J Cancer 97: 458–465, 2002.

    Article  PubMed  CAS  Google Scholar 

  29. Okhrimenko H, Lu W, Xiang C, et al: Protein kinase C-epsilon regulates the apoptosis and survival of glioma cells. Cancer Res 65: 7301–7309, 2005.

    Article  PubMed  CAS  Google Scholar 

  30. Okhrimenko H, Lu W, Xiang C, et al: Roles of tyrosine phosphorylation and cleavage of protein kinase Cdelta in its protective effect against tumor necrosis factor-related apoptosis inducing ligand-induced apoptosis. J Biol Chem 280: 23643–23652, 2005.

    Article  PubMed  CAS  Google Scholar 

  31. Pardo OE, Wellbrock C, Khanzada UK, et al: FGF-2 protects small cell lung cancer cells from apoptosis through a complex involving PKCepsilon, B-Raf and S6K2. EMBO J 25: 3078–3088, 2006.

    Article  PubMed  CAS  Google Scholar 

  32. Platzbecker U, Ward JL, Deeg HJ: Chelerythrin activates caspase-8, downregulates FLIP long and short, and overcomes resistance to tumour necrosis factor-related apoptosis-inducing ligand in KG1a cells. Br J Haematol 122: 489–497, 2003.

    Article  PubMed  CAS  Google Scholar 

  33. Reed JC: Drug insight: Cancer therapy strategies based on restoration of endogenous cell death mechanisms. Nat Clin Pract Oncol 3: 388–398, 2006.

    Article  PubMed  CAS  Google Scholar 

  34. Saraiva L, Fresco P, Pinto E, et al: Isoform-selectivity of PKC inhibitors acting at the regulatory and catalytic domain of mammalian PKC-alpha, -betaI, -delta,-eta and -zeta. J Enzyme Inhib Med Chem 18: 475–483, 2003.

    Article  PubMed  CAS  Google Scholar 

  35. Sarker M, Ruiz-Ruiz C, Lopez-Rivas A: Activation of protein kinase C inhibits TRAIL-induced caspases activation, mitochondrial events and apoptosis in a human leukemic T cell line. Cell Death Differ 8: 172–181, 2001.

    Article  PubMed  CAS  Google Scholar 

  36. Schiller JH, Harrington D, Belani CP, et al: Comparison of four chemotherapy regimens for advanced non-small-cell lung cancer. N Engl J Med 346: 92–98, 2002.

    Article  PubMed  CAS  Google Scholar 

  37. Shi RX, Ong CN, Shen HM: Protein kinase C inhibition and x-linked inhibitor of apoptosis protein degradation contribute to the sensitization effect of luteolin on tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in cancer cells. Cancer Res 65: 7815–7823, 2005.

    Article  PubMed  CAS  Google Scholar 

  38. Shimomoto H, Hasegawa Y, Nozaki Y, et al: Expression of tumor necrosis factor receptors in human lung cancer cells and normal lung tissues. Am J Respir Cell Mol Biol 13: 271–278, 1995.

    PubMed  CAS  Google Scholar 

  39. Shinohara H, Kayagaki N, Yagita H, et al: A protective role of PKCepsilon against TNF-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in glioma cells. Biochem Biophys Res Commun 284: 1162–1167, 2001.

    Article  PubMed  CAS  Google Scholar 

  40. Sivaprasad U, Shankar E, Basu A: Downregulation of Bid is associated with PKCepsilon-mediated TRAIL resistance. Cell Death Differ 14: 851–860, 2007.

    Article  PubMed  CAS  Google Scholar 

  41. Sonnemann J, Gange J, Kumar KS, et al: Histone deacetylase inhibitors interact synergistically with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to induce apoptosis in carcinoma cell lines. Invest New Drugs 23: 99–109, 2005.

    Article  PubMed  CAS  Google Scholar 

  42. Sonnemann J, Gekeler V, Sagrauske A, et al: Down-regulation of protein kinase Ceta potentiates the cytotoxic effects of exogenous tumor necrosis factor-related apoptosis-inducing ligand in PC-3 prostate cancer cells. Mol Cancer Ther 3: 773–781, 2004.

    PubMed  CAS  Google Scholar 

  43. Spierings DC, de Vries EG, Timens W, et al: Expression of TRAIL and TRAIL death receptors in stage III non-small cell lung cancer tumors. Clin Cancer Res 9: 3397–3405, 2003.

    PubMed  CAS  Google Scholar 

  44. Sullivan JP, Connor JR, Shearer BG, et al: 2,6-Diamino-N-([1-(1-oxotridecyl)-2-piperidinyl] methyl)hexanamide (NPC 15437): a novel inhibitor of protein kinase C interacting at the regulatory domain. Mol Pharmacol 41: 38–44, 1992.

    PubMed  CAS  Google Scholar 

  45. Tillman DM, Izeradjene K, Szucs KS, et al: Rottlerin sensitizes colon carcinoma cells to tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis via uncoupling of the mitochondria independent of protein kinase C. Cancer Res 63: 5118–5125, 2003.

    PubMed  CAS  Google Scholar 

  46. Trauzold A, Wermann H, Arlt A, et al: CD95 and TRAIL receptor-mediated activation of protein kinase C and NF-kappaB contributes to apoptosis resistance in ductal pancreatic adenocarcinoma cells. Oncogene 20: 4258–4269, 2001.

    Article  PubMed  CAS  Google Scholar 

  47. van Valen F, Fulda S, Schafer KL, et al: Selective and nonselective toxicity of TRAIL/Apo2L combined with chemotherapy in human bone tumour cells vs. normal human cells. Int J Cancer 107: 929–940, 2003.

    Article  PubMed  CAS  Google Scholar 

  48. Wu D, Thakore CU, Wescott GG, et al: Integrin signaling links protein kinase Cepsilon to the protein kinase B/Akt survival pathway in recurrent prostate cancer cells. Oncogene 23: 8659–8672, 2004.

    Article  PubMed  CAS  Google Scholar 

  49. Zhang J, Liu N, Zhang J, et al: PKCdelta protects human breast tumor MCF-7 cells against tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis. J Cell Biochem 96: 522–532, 2005.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Research Center of Pharmacology and Experimental Therapeutics, Ernst Moritz Arndt University, Greifswald, Germany

    Matthias Felber & Jürgen Sonnemann

  2. Department of Pediatric Oncology/Hematology, Ernst Moritz Arndt University, Greifswald, Germany

    James F Beck

Authors
  1. Matthias Felber
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jürgen Sonnemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. James F Beck
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to James F Beck.

Additional information

This work was supported by a grant from the “Wilhelm Sander-Stiftung, Neustadt/Donau”.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Felber, M., Sonnemann, J. & Beck, J.F. Inhibition of novel protein kinase Cɛ augments TRAIL-induced cell death in A549 lung cancer cells. Pathol. Oncol. Res. 13, 295–301 (2007). https://doi.org/10.1007/BF02940308

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02940308

Key words

Search

Navigation