Skip to main content

Advertisement

SpringerLink
Log in

The role of tissue transglutaminase (TG2) in regulating the tumour progression of the mouse colon carcinoma CT26

  • Original Article
  • Published:
Amino Acids Aims and scope Submit manuscript

Abstract

The multifunctional enzyme tissue transglutaminase (TG2) is reported to both mediate and inhibit tumour progression. To elucidate these different roles of TG2, we established a series of stable-transfected mouse colon carcinoma CT26 cells expressing a catalytically active (wild type) and a transamidating-inactive TG2 (Cys277Ser) mutant. Comparison of the TG2-transfected cells with the empty vector control indicated no differences in cell proliferation, apoptosis and susceptibility to doxorubicin, which correlated with no detectable changes in the activation of the transcription factor NF-κB. TG2-transfected cells showed increased expression of integrin β3, and were more adherent and less migratory on fibronectin than control cells. Direct interaction of TG2 with β3 integrins was demonstrated by immunoprecipitation, suggesting that TG2 acts as a coreceptor for fibronectin with β3 integrins. All cells expressed the same level of TGFβ receptors I and II, but only cells transfected with active TG2 had increased levels of TGFβ1 and matrix-deposited fibronectin, which could be inhibited by TG2 site-directed inhibitors. Moreover, only cells transfected with active TG2 were capable of inhibiting tumour growth when compared to the empty vector controls. We conclude that in this colon carcinoma model increased levels of active TG2 are unfavourable to tumour growth due to their role in activation of TGFβ1 and increased matrix deposition, which in turn favours increased cell adhesion and a lowered migratory and invasive behaviour.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Ai LB, Kim WJ, Demircan B, Dyer LM, Bray KJ, Skehan RR, Massoll NA, Brown KD (2008) The transglutaminase 2 gene (TGM2), a potential molecular marker for chemotherapeutic drug sensitivity, is epigenetically silenced in breast cancer. Carcinogenesis 29:510–518

    Article  PubMed  CAS  Google Scholar 

  • Akimov SS, Belkin AM (2001) Cell-surface transglutaminase promotes fibronectin assembly via interaction with the gelatin-binding domain of fibronectin: a role in TGF beta-dependent matrix deposition. J Cell Sci 114:2989–3000

    PubMed  CAS  Google Scholar 

  • Akimov SS, Belkin AM (2003) Opposing roles of Ras/Raf oncogenes and the MEK1/ERK signaling module in regulation of expression and adhesive function of surface transglutaminase. J Biol Chem 278:35609–35619

    Article  PubMed  CAS  Google Scholar 

  • Akimov SS, Krylov D, Fleischman LF, Belkin AM (2000) Tissue transglutaminase is an integrin-binding adhesion coreceptor for fibronectin. J Cell Biol 148:825–838

    Article  PubMed  CAS  Google Scholar 

  • Antonyak MA, Li B, Regan AD, Feng Q, Dusaban SS, Cerione RA (2009) Tissue transglutaminase is an essential participant in the epidermal growth factor-stimulated signaling pathway leading to cancer cell migration and invasion. J Biol Chem 284:17914–17925

    Article  PubMed  CAS  Google Scholar 

  • Balklava Z, Verderio E, Collighan R, Gross S, Adams J, Griffin M (2002) Analysis of tissue transglutaminase function in the migration of Swiss 3T3 fibroblasts: the active-state conformation of the enzyme does not affect cell motility but is important for its secretion. J Biol Chem 277:16567–16575

    Article  PubMed  CAS  Google Scholar 

  • Barnes RN, Bungay PJ, Elliott BM, Walton PL, Griffin M (1985) Alterations in the distribution and activity of transglutaminase during tumour growth and metastasis. Carcinogenesis 6:459–463

    Article  PubMed  CAS  Google Scholar 

  • Belkin AM, Zemskov EA, Hang J, Akimov SS, Sikora S, Strongin AY (2004) Cell-surface-associated tissue transglutaminase is a target of MMP-2 proteolysis. Biochemistry 43:11760–11769

    Article  PubMed  CAS  Google Scholar 

  • Chau DYS, Collighan RJ, Verderio EAM, Addy VL, Griffin M (2005) The cellular response to transglutaminase-cross-linked collagen. Biomaterials 26:6518–6529

    Article  PubMed  CAS  Google Scholar 

  • Collighan RJ, Griffin M (2009) Transglutaminase 2 cross-linking of matrix proteins: biological significance and medical applications. Amino Acids 36:659–670

    Article  PubMed  CAS  Google Scholar 

  • Di Giacomo G, Lentini A, Beninati S, Piacentini M, Rodolfo C (2009) In vivo evaluation of type 2 transglutaminase contribution to the metastasis formation in melanoma. Amino Acids 36:717–724

    Article  PubMed  CAS  Google Scholar 

  • Fisher M, Jones RA, Huang L, Haylor JL, El Nahas M, Griffin M, Johnson TS (2009) Modulation of tissue transglutaminase in tubular epithelial cells alters extracellular matrix levels: a potential mechanism of tissue scarring. Matrix Biol 28:20–31

    Article  PubMed  CAS  Google Scholar 

  • Gaudry CA, Verderio E, Aeschlimann D, Cox A, Smith C, Griffin M (1999) Cell surface localization of tissue transglutaminase is dependent on a fibronectin-binding site in its N-terminal beta-sandwich domain. J Biol Chem 274:30707–30714

    Article  PubMed  CAS  Google Scholar 

  • Griffin M, Casadio R, Bergamini CM (2002) Transglutaminases: nature’s biological glues. Biochem J 368:377–396

    Article  PubMed  CAS  Google Scholar 

  • Griffin M, Mongeot A, Collighan R, Saint RE, Jones RA, Coutts IGC, Rathbone DL (2008) Synthesis of potent water-soluble tissue transglutaminase inhibitors. Bioorg Med Chem Lett 18:5559–5562

    Article  PubMed  CAS  Google Scholar 

  • Grigoriev MY, Suspitsin EN, Togo AV, Pozharisski KM, Ivanova OA, Nardacci R, Falasca L, Piacentini M, Imyanitov EN, Hanson KP (2001) Tissue transglutaminase expression in breast carcinomas. J Exp Clin Cancer Res 20:265–268

    PubMed  CAS  Google Scholar 

  • Gundemir S, Johnson GV (2009) Intracellular localization and conformational state of transglutaminase 2: implications for cell death. PLoS One 4:e6123

    Article  PubMed  Google Scholar 

  • Hand D, Elliott BM, Griffin M (1988) Expression of the cytosolic and particulate forms of transglutaminase during chemically induced rat liver carcinogenesis. Biochim Biophys Acta 970:137–145

    Article  PubMed  CAS  Google Scholar 

  • Herman JF, Mangala LS, Mehta K (2006) Implications of increased tissue transglutaminase (TG2) expression in drug-resistant breast cancer (MCF-7) cells. Oncogene 25:3049–3058

    Article  PubMed  CAS  Google Scholar 

  • Huang LH, Haylor JL, Hau Z, Jones RA, Vickers ME, Wagner B, Griffin M, Saint RE, Coutts IGC, El Nahas AM, Johnson TS (2009) Transglutaminase inhibition ameliorates experimental diabetic nephropathy. Kidney Int 76:383–394

    Article  PubMed  CAS  Google Scholar 

  • Ientile R, Caccamo D, Griffin M (2007) Tissue transglutaminase and the stress response. Amino Acids 33:385–394

    Article  PubMed  CAS  Google Scholar 

  • Johnson TS, Knight CR, el-Alaoui S, Mian S, Rees RC, Gentile V, Davies PJ, Griffin M (1994) Transfection of tissue transglutaminase into a highly malignant hamster fibrosarcoma leads to a reduced incidence of primary tumour growth. Oncogene 9:2935–2942

    PubMed  CAS  Google Scholar 

  • Johnson TS, Fisher M, Haylor JL, Hau Z, Skill NJ, Jones R, Saint R, Coutts I, Vickers ME, El Nahas AM, Griffin M (2007) Transglutaminase inhibition reduces fibrosis and preserves function in experimental chronic kidney disease. J Am Soc Nephrol 18:3078–3088

    Article  PubMed  CAS  Google Scholar 

  • Jones RA, Nicholas B, Mian S, Davies PJ, Griffin M (1997) Reduced expression of tissue transglutaminase in a human endothelial cell line leads to changes in cell spreading, cell adhesion and reduced polymerisation of fibronectin. J Cell Sci 110:2461–2472

    PubMed  CAS  Google Scholar 

  • Jones RA, Kotsakis P, Johnson TS, Chau DY, Ali S, Melino G, Griffin M (2006) Matrix changes induced by transglutaminase 2 lead to inhibition of angiogenesis and tumor growth. Cell Death Differ 13:1442–1453

    Article  PubMed  CAS  Google Scholar 

  • Knight CR, Rees RC, Griffin M (1990) Reduction in transglutaminase activity associated with tumour metastasis is due to the presence of an inactive form of the enzyme. Biochem Soc Trans 18:681–682

    PubMed  CAS  Google Scholar 

  • Kotsakis P, Griffin M (2007) Tissue transglutaminase in tumour progression: friend or foe? Amino Acids 33:373–384

    Article  PubMed  CAS  Google Scholar 

  • Lentini A, Abbruzzese A, Caraglia M, Marra M, Beninati S (2004) Protein-polyamine conjugation by transglutaminase in cancer cell differentiation: review article. Amino Acids 26:331–337

    Article  PubMed  CAS  Google Scholar 

  • Lorand L, Campbell-Wilkes LK, Cooperstein L (1972) A filter paper assay for transamidating enzymes using radioactive amine substrates. Anal Biochem 50:623–631

    Article  PubMed  CAS  Google Scholar 

  • Mangala LS, Arun B, Sahin AA, Mehta K (2005) Tissue transglutaminase-induced alterations in extracellular matrix inhibit tumor invasion. Mol Cancer 4:1–8

    Article  Google Scholar 

  • Mann AP, Verma A, Sethi G, Manavathi B, Wang H, Fok JY, Kunnumakkara AB, Kumar R, Aggarwal BB, Mehta K (2006) Overexpression of tissue transglutaminase leads to constitutive activation of nuclear factor-kappaB in cancer cells: delineation of a novel pathway. Cancer Res 66:8788–8795

    Article  PubMed  CAS  Google Scholar 

  • Mehta K (2009) Biological and therapeutic significance of tissue transglutaminase in pancreatic cancer. Amino Acids 36:709–716

    Article  PubMed  CAS  Google Scholar 

  • Myrsky E, Caja S, Simon-Vecsei Z, Korponay-Szabo IR, Nadalutti C, Collighan R, Mongeot A, Griffin M, Maki M, Kaukinen K, Lindfors K (2009) Celiac disease IgA modulates vascular permeability in vitro through the activity of transglutaminase 2 and RhoA. Cell Mol Life Sci 66:3375–3385

    Article  PubMed  CAS  Google Scholar 

  • Nicholas B, Smethurst P, Verderio E, Jones R, Griffin M (2003) Cross-linking of cellular proteins by tissue transglutaminase during necrotic cell death: a mechanism for maintaining tissue integrity. Biochem J 371:413–422

    Article  PubMed  CAS  Google Scholar 

  • Piacentini M, Piredda L, Starace DT, Annicchiarico-Petruzzelli M, Mattei M, Oliverio S, Farrace MG, Melino G (1996) Differential growth of N- and S-type human neuroblastoma cells xenografted into scid mice. Correlation with apoptosis. J Pathol 180:415–422

    Article  PubMed  CAS  Google Scholar 

  • Rosenkranz S (2004) TGF-beta1 and angiotensin networking in cardiac remodeling. Cardiovasc Res 63:423–432

    Article  PubMed  CAS  Google Scholar 

  • Shweke N, Boulos N, Jouanneau C, Vandermeersch S, Melino G, Dussaule JC, Chatziantoniou C, Ronco P, Boffa JJ (2008) Tissue transglutaminase contributes to interstitial renal fibrosis by favoring accumulation of fibrillar collagen through TGF-beta activation and cell infiltration. Am J Pathol 173:631–642

    Article  PubMed  CAS  Google Scholar 

  • Smethurst PA, Griffin M (1996) Measurement of tissue transglutaminase activity in a permeabilized cell system: its regulation by Ca2+ and nucleotides. Biochem J 313:803–808

    PubMed  CAS  Google Scholar 

  • Switala-Jelen K, Dabrowska K, Opolski A, Lipinska L, Nowaczyk M, Gorski A (2004) The biological functions of beta3 integrins. Folia Biol (Praha) 50:143–152

    CAS  Google Scholar 

  • Tadokoro S, Shattil SJ, Eto K, Tai V, Liddington RC, de Pereda JM, Ginsberg MH, Calderwood DA (2003) Talin binding to integrin beta tails: a final common step in integrin activation. Science 302:103–106

    Article  PubMed  CAS  Google Scholar 

  • Telci D, Wang Z, Li X, Verderio EA, Humphries MJ, Baccarini M, Basaga H, Griffin M (2008) Fibronectin-tissue transglutaminase matrix rescues RGD-impaired cell adhesion through syndecan-4 and beta1 integrin co-signaling. J Biol Chem 283:20937–20947

    Article  PubMed  CAS  Google Scholar 

  • Telci D, Colligha RJ, Basaga H, Griffin M (2009) Increased TG2 expression can result in induction of transforming growth factor beta 1, causing increased synthesis and deposition of matrix proteins, which can be regulated by nitric oxide. J Biol Chem 284:29547–29558

    Article  PubMed  CAS  Google Scholar 

  • Toth B, Sarang Z, Vereb G, Zhang AL, Tanaka S, Melino G, Fesus L, Szondy Z (2009) Over-expression of integrin beta 3 can partially overcome the defect of integrin beta 3 signaling in transglutaminase 2 null macrophages. Immunol Lett 126:22–28

    Article  PubMed  CAS  Google Scholar 

  • Verderio E, Nicholas B, Gross S, Griffin M (1998) Regulated expression of tissue transglutaminase in Swiss 3T3 fibroblasts: effects on the processing of fibronectin, cell attachment, and cell death. Exp Cell Res 239:119–138

    Article  PubMed  CAS  Google Scholar 

  • Verderio E, Gaudry C, Gross S, Smith C, Downes S, Griffin M (1999) Regulation of cell surface tissue transglutaminase: effects on matrix storage of latent transforming growth factor-beta binding protein-1. J Histochem Cytochem 47:1417–1432

    Article  PubMed  CAS  Google Scholar 

  • Verderio EA, Telci D, Okoye A, Melino G, Griffin M (2003) A novel RGD-independent cell adhesion pathway mediated by fibronectin-bound tissue transglutaminase rescues cells from anoikis. J Biol Chem 278:42604–42614

    Article  PubMed  CAS  Google Scholar 

  • Verma A, Mehta K (2007a) Tissue transglutaminase-mediated chemoresistance in cancer cells. Drug Resist Update 10:144–151

    Article  CAS  Google Scholar 

  • Verma A, Mehta K (2007b) Transglutaminase-mediated activation of nuclear transcription factor-kappa B in cancer cells: a new therapeutic opportunity. Curr Cancer Drug Targets 7:559–565

    Article  PubMed  CAS  Google Scholar 

  • Yuan L, Siegel M, Choi K, Khosla C, Miller CR, Jackson EN, Piwnica-Worms D, Rich KM (2007) Transglutaminase 2 inhibitor, KCC009, disrupts fibronectin assembly in the extracellular matrix and sensitizes orthotopic glioblastomas to chemotherapy. Oncogene 26:2563–2573

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We would thank Dr. Elisabetta A.M. Verderio for the helpful advice on the molecular biology aspects of the project.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, B47ET, United Kingdom

    Panayiotis Kotsakis, Zhuo Wang, Russell John Collighan & Martin Griffin

  2. Department of Life Sciences, Nottingham Trent University, Nottingham, NG11 8NS, United Kingdom

    Panayiotis Kotsakis

Authors
  1. Panayiotis Kotsakis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhuo Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Russell John Collighan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Martin Griffin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Martin Griffin.

Additional information

P. Kotsakis and Z. Wang contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kotsakis, P., Wang, Z., Collighan, R.J. et al. The role of tissue transglutaminase (TG2) in regulating the tumour progression of the mouse colon carcinoma CT26. Amino Acids 41, 909–921 (2011). https://doi.org/10.1007/s00726-010-0790-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00726-010-0790-1

Keywords

Search

Navigation