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Investigation of the mechanisms of tissue factor-mediated evasion of tumour cells from cellular cytotoxicity

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Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Aims

We previously reported that overexpression of tissue factor (TF) protected HT29 tumour cells from cellular cytotoxicity through a mechanism requiring the presence of the cytoplasmic domain of TF. In this investigation the mechanism of TF-mediated immune evasion has been examined.

Methods

The influence of alanine-substitution at Ser253 and Ser258 of TF (TFAla253 and TF Ala258) on the induction of cytotoxic evasion, as well as expression of vascular cell adhesion molecule-1 and intra-cellular adhesion molecule-1 (VCAM-1 and ICAM-1) was investigated. Moreover, we examined the effect of transfection of four 20-mer peptides, corresponding to the C-terminal residues of TF, with different phosphorylation states, on promotion of evasion from cell cytotoxicity.

Results

Cells overexpressing TFAla258 and to a lesser extent overexpressing TFAla253, exhibited a reduced ability to evade cellular cytotoxicity compared to cells overexpressing the wild-type TF. Furthermore, the increase in protection acquired was greatest on transfection of Ser258-phosphsorylated form of the cytoplasmic peptide, lower in double-phosphorylated and Ser253-phosphorylated peptides respectively, and lowest in the unphosphorylated form. Finally, the expression of VCAM-1 mRNA as well as surface antigen was reduced on overexpression of TFwt but was partially reverted in the cells transfected to overexpress TFAla253 or TFAla258.

Conclusions

These data show that the phosphorylation of TF at Ser258 and to a lesser extent Ser253, plays an essential role in the protective influence of TF on immune evasion by tumour cells, and that the mechanism could involve the downregulation of key surface antigens, such as adhesion proteins, involved in cell:cell interaction.

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References

  1. Shoji M, Hancock WW, Abe K, Micko C, Casper KA, Baine RM, Wilcox JN, Danave I, Dillehay DL, Matthews E, Contrino J, Morrissey JH, Gordon S, Edgington TS, Kudryk B, Kreutzer DL, Rickles FR (1998) Activation of coagulation and angiogenesis in cancer: immunohistochemical localization in situ of clotting proteins and vascular endothelial growth factor in human cancer. Am J Pathol 152:399–411

    PubMed  CAS  Google Scholar 

  2. Ruf W, Mueller BM (2006) Thrombin generation and the pathogenesis of cancer. Semin Thromb Hemost 32:61–68

    Article  PubMed  CAS  Google Scholar 

  3. Versteeg HH, Spek CA, Peppelenbosch MP, Richel DJ (2004) Tissue factor and cancer metastasis: the role of intracellular and extracellular signaling pathways. Mol Med 10:6–11

    Article  PubMed  CAS  Google Scholar 

  4. Belting M, Dorrell MI, Sandgren S, Aguilar E, Ahamed J, Dorfleutner A, Carmeliet P, Mueller BM, Friedlander M, Ruf W (2004) Regulation of angiogenesis by tissue factor cytoplasmic domain signaling. Nat Med 10:502–509

    Article  PubMed  CAS  Google Scholar 

  5. Versteeg HH, Peppelenbosch MP, Spek CA (2003) Tissue factor signal transduction in angiogenesis. Carcinogenesis 24:1009–1013

    Article  PubMed  CAS  Google Scholar 

  6. Zhang Y, Deng Y, Luther T, Muller M, Ziegler R, Waldherr R, Stern DM, Nawroth PP (1994) Tissue factor controls the balance of angiogenic and antiangiogenic properties of tumor cells in mice. J Clin Invest 94:1320–1327

    Article  PubMed  CAS  Google Scholar 

  7. Bromberg ME, Konigsberg WH, Madison JF, Pawashe A, Garen A (1995) Tissue factor promotes melanoma metastasis by a pathway independent of blood coagulation. Proc Natl Acad Sci USA 92:8205–8209

    Article  PubMed  CAS  Google Scholar 

  8. Abe K, Shoji M, Chen J, Bierhaus A, Danave I, Micko C, Casper K, Dillehay DL, Nawroth PP, Rickles FR (1999) Regulation of vascular endothelial growth factor production and angiogenesis by the cytoplasmic tail of tissue factor. Proc Natl Acad Sci USA 96:8663–8668

    Article  PubMed  CAS  Google Scholar 

  9. Bromberg ME, Sundaram R, Homer RJ, Garen A, Konigsberg WH (1999) Role of tissue factor in metastasis: functions of the cytoplasmic and extracellular domains of the molecule. Thromb Haemost 82:88–92

    PubMed  CAS  Google Scholar 

  10. Zioncheck TF, Roy S, Vehar GA (1992) The cytoplasmic domain of tissue factor is phosphorylated by a protein kinase C-dependent mechanism. J Biol Chem 267:3561–3564

    PubMed  CAS  Google Scholar 

  11. Mody RS, Carson SD (1997) Tissue factor cytoplasmic domain peptide is multiply phosphorylated in vitro. Biochemistry 36:7869–7875

    Article  PubMed  CAS  Google Scholar 

  12. Bromberg ME, Sundaram R, Homer RJ, Garen A, Konigsberg WH (1999) Role of tissue factor in metastasis: functions of the cytoplasmic and extracellular domains of the molecule. Thromb Haemost 82:88–92

    PubMed  CAS  Google Scholar 

  13. Ruf W (2001) Molecular regulation of blood clotting in tumor biology. Haemostasis 31:5–7

    PubMed  CAS  Google Scholar 

  14. Li C, Colman LM, Collier MEW, Dyer CE, Greenman J, Ettelaie C (2006) Tumour-expressed tissue factor inhibits cellular cytotoxicity. Cancer Immunol Immunother 55:1301–1308

    Article  PubMed  CAS  Google Scholar 

  15. Weiss L (1990) Metastatic inefficiency. Adv Cancer Res 54:159–211

    Article  PubMed  CAS  Google Scholar 

  16. Budinsky AC, Brodowicz T, Wiltschke C, Czerwenk K, Michil I, Krainer M, Zielinski CC (1997) Decreased expression of ICAM-1 and its induction by tumour necrosis factor on breast cancer cells in vitro. Int J Cancer 71:1086–1090

    Article  PubMed  CAS  Google Scholar 

  17. Melder RJ, Koenig GC, Munn LL, Jain RK (1997) Adhesion of activated natural killer cells to tumor necrosis factor-alpha-treated endothelium under physiological flow conditions. Nat Immun 15:154–163

    CAS  Google Scholar 

  18. Huang M, Syed R, Stura EA, Stone MJ, Stefanko RS, Ruf W, Edgington TS, Wilson IA (1998) The mechanism of an inhibitory antibody on TF-initiated blood coagulation revealed by the crystal structures of human tissue factor, Fab 5G9 and TF.G9 complex. J Mol Biol 275:873–94

    Article  PubMed  CAS  Google Scholar 

  19. Angell YM, Alsina J, Albericio F, Barany G (2002) Practical protocols for stepwise solid-phase synthesis of cysteine-containing peptides. J Pept Res 60:292–299

    Article  PubMed  CAS  Google Scholar 

  20. Ettelaie C, Adam JM, James NJ, Oke AO, Harrison JA, Bunce TD, Bruckdorfer KR (1999) The role of C-terminal domain in the inhibitory functions of tissue factor pathway inhibitor. FEBS Lett 463:341–344

    Article  PubMed  CAS  Google Scholar 

  21. Bustin SA (2000) Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol 25:169–193

    Article  PubMed  CAS  Google Scholar 

  22. Matsuno H, Stassen JM, Vermylen J, Deckmyn H (1994) Inhibition of integrin function by a cyclic RGD-containing peptide prevents neointima formation. Circulation 90:2203–2206

    PubMed  CAS  Google Scholar 

  23. Gurrath M, Muller G, Kessler H, Aumailley M, Timpl R (1992) Conformation/activity studies of rationally designed potent anti-adhesive RGD peptides. Eur J Biochem 210:911–921

    Article  PubMed  CAS  Google Scholar 

  24. Miyata R, Iwabuchi K, Watanabe S, Sato N, Nagaoka I (1999) Exposure of intestinal epithelial cell HT29 to bile acids and ammonia enhances Mac-1-mediated neutrophil adhesion. Inflamm Res 48:265–273

    Article  PubMed  CAS  Google Scholar 

  25. Dorfleutner A, Ruf W (2003) Regulation of tissue factor cytoplasmic domain phosphorylation by palmitoylation. Blood 102:3998–4005

    Article  PubMed  CAS  Google Scholar 

  26. Siegbahn A (2000) Cellular consequences upon factor VIIa binding to tissue factor. Haemostasis 30:41–47

    Article  PubMed  CAS  Google Scholar 

  27. Morris DR, Ding Y, Ricks TK, Gullapalli A, Wolfe BL, Trejo J (2006) Protease-activated receptor-2 is essential for factor VIIa and Xa-induced signaling, migration, and invasion of breast cancer cells. Cancer Res 66:307–314

    Article  PubMed  CAS  Google Scholar 

  28. Bromberg ME, Bailly MA, Konigsberg WH (2001) Role of protease-activated receptor 1 in tumor metastasis promoted by tissue factor. Thromb Haemost 86:1210–1214

    PubMed  CAS  Google Scholar 

  29. Ahamed J, Ruf W (2004) Protease-activated receptor 2-dependent phosphorylation of the tissue factor cytoplasmic domain. J Biol Chem 279:23038–23044

    Article  PubMed  CAS  Google Scholar 

  30. Mueller B, Ruf W (1998) Requirement for binding of catalytically active factor VIIa in tissue factor-dependent experimental metastasis. J Clin Invest 101:1372–1378

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Biomedical Section, Department of Biological Sciences, University of Hull, Cottingham Road, Hull, HU6 7RX, UK

    Chao Li, Mary E. W. Collier, G. Alkisitis Frentzou & Camille Ettelaie

  2. Cancer Division, Postgraduate Medical Institute, University of Hull, Cottingham Road, Hull, HU6 7RX, UK

    John Greenman

Authors
  1. Chao Li
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  2. Mary E. W. Collier
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  3. G. Alkisitis Frentzou
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  4. John Greenman
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  5. Camille Ettelaie
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Correspondence to Camille Ettelaie.

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Li, C., Collier, M.E.W., Frentzou, G.A. et al. Investigation of the mechanisms of tissue factor-mediated evasion of tumour cells from cellular cytotoxicity. Cancer Immunol Immunother 57, 1347–1355 (2008). https://doi.org/10.1007/s00262-008-0469-6

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  • DOI: https://doi.org/10.1007/s00262-008-0469-6

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