Initiation of human colon cancer cell proliferation by trypsin acting at protease-activated receptor-2

Abstract

The protease-activated receptor-2 (PAR-2) is a G protein-coupled receptor that is cleaved and activated by trypsin. We investigated the expression of PAR-2 and the role of trypsin in cell proliferation in human colon cancer cell lines. A total of 10 cell lines were tested for expression of PAR-2 mRNA by Northern blot and RT-PCR. PAR-2 protein was detected by immunofluorescence. Trypsin and the peptide agonist SLIGKV (AP2) were tested for their ability to induce calcium mobilization and to promote cell proliferation on serum-deprived cells. PAR-2 mRNA was detected by Northern blot analysis in 6 out of 10 cell lines [HT-29, Cl.19A, Caco-2, SW480, HCT-8 and T84]. Other cell lines expressed low levels of transcripts, which were detected only by RT-PCR. Further results were obtained with HT-29 cells: (1) PAR-2 protein is expressed at the cell surface; (2) an increase in intracellular calcium concentration was observed upon trypsin (1-100 nM) or AP2 (10-100 microM) challenges; (3) cells grown in serum-deprived media supplemented with trypsin (0.1-1 nM) or AP2 (1-300 microM) exhibited important mitogenic responses (3-fold increase of cell number). Proliferative effects of trypsin or AP2 were also observed in other cell lines expressing PAR-2. These data show that subnanomolar concentrations of trypsin, acting at PAR-2, promoted the proliferation of human colon cancer cells. The results of this study indicate that trypsin could be considered as a growth factor and unravel a new mechanism whereby serine proteases control colon tumours.

References 55

1. 1.

   Augeron C., Laboisse C. L. Emergence of permanently differentiated cell clones in a human colonic cancer cell line in culture after treatment with sodium butyrate. Cancer Res. 1984;44(9):3961-3969.

   Not in our library

2. 2.

   Belham C. M., Tate R. J., Scott P. H., Pemberton A. D., Miller H. R., Wadsworth R. M., Gould G. W., Plevin R. Trypsin stimulates proteinase-activated receptor-2-dependent and -independent activation of mitogen-activated protein kinases. Biochem J. 1996;320(Pt 3):939-946.

   Not in our library10.1042/bj3200939 (opens in a new tab)

3. 3.

   Bernard-Perrone F., Carrere J., Renaud W., Moriscot C., Thoreux K., Bernard P., Servin A., Balas D., Senegas-Balas F. Pancreatic trypsinogen I expression during cell growth and differentiation of two human colon carcinoma cells. Am J Physiol. 1998;274(6 Pt 1):G1077-G1086.

   Not in our library10.1152/ajpgi.1998.274.6.g1077 (opens in a new tab)

4. 4.

   Blackhart B. D., Emilsson K., Nguyen D., Teng W., Martelli A. J., Nystedt S., Sundelin J., Scarborough R. M. Ligand cross-reactivity within the protease-activated receptor family. J Biol Chem. 1996;271(28):16466-16471.

   Not in our library10.1074/jbc.271.28.16466 (opens in a new tab)

5. 5.

   Blasi F., Stoppelli M. P. Proteases and cancer invasion: from belief to certainty. AACR meeting on proteases and protease inhibitors in cancer, Nyborg, Denmark, 14-18 June 1998. Biochim Biophys Acta. 1999;1423(1):R35-R44.

   Not in our library10.1016/s0304-419x(98)00034-1 (opens in a new tab)

6. 6.

   Blumberg P. M., Robbins P. W. Effect of proteases on activation of resting chick embryo fibroblasts and on cell surface proteins. Cell. 1975;6(2):137-147.

   Not in our library10.1016/0092-8674(75)90004-5 (opens in a new tab)

7. 7.

   Bohm S. K., Kong W., Bromme D., Smeekens S. P., Anderson D. C., Connolly A., Kahn M., Nelken N. A., Coughlin S. R., Payan D. G. Molecular cloning, expression and potential functions of the human proteinase-activated receptor-2. Biochem J. 1996;314(Pt 3):1009-1016.

   Not in our library10.1042/bj3141009 (opens in a new tab)

8. 8.

   Bustos D., Tiscornia O., Caldarini M. I., Negri G., Pons S., Ogawa K., De Paula J. A. Colonic proteolysis following pancreatic duct ligation in the rat. Int J Pancreatol. 1994;16(1):45-49.

   Not in our library10.1007/bf02925609 (opens in a new tab)

9. 9.

   Böhm S. K., Khitin L. M., Grady E. F., Aponte G., Payan D. G., Bunnett N. W. Mechanisms of desensitization and resensitization of proteinase-activated receptor-2. J Biol Chem. 1996;271(36):22003-22016.

   Not in our library10.1074/jbc.271.36.22003 (opens in a new tab)

10. 10.

    Carney D. H., Cunningham D. D. Initiation of check cell division by trypsin action at the cell surface. Nature. 1977;268(5621):602-606.

    Not in our library10.1038/268602a0 (opens in a new tab)

11. 11.

    Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979;18(24):5294-5299.

    Not in our library10.1021/bi00591a005 (opens in a new tab)

12. 12.

    D'Andrea M. R., Derian C. K., Leturcq D., Baker S. M., Brunmark A., Ling P., Darrow A. L., Santulli R. J., Brass L. F., Andrade-Gordon P. Characterization of protease-activated receptor-2 immunoreactivity in normal human tissues. J Histochem Cytochem. 1998;46(2):157-164.

    Not in our library10.1177/002215549804600204 (opens in a new tab)

13. 13.

    Darmoul D., Lacasa M., Baricault L., Marguet D., Sapin C., Trotot P., Barbat A., Trugnan G. Dipeptidyl peptidase IV (CD 26) gene expression in enterocyte-like colon cancer cell lines HT-29 and Caco-2. Cloning of the complete human coding sequence and changes of dipeptidyl peptidase IV mRNA levels during cell differentiation. J Biol Chem. 1992;267(7):4824-4833.

    Not in our library

14. 14.

    DeFea K. A., Zalevsky J., Thoma M. S., Déry O., Mullins R. D., Bunnett N. W. beta-arrestin-dependent endocytosis of proteinase-activated receptor 2 is required for intracellular targeting of activated ERK1/2. J Cell Biol. 2000;148(6):1267-1281.

    Not in our library10.1083/jcb.148.6.1267 (opens in a new tab)

15. 15.

    Déry O., Corvera C. U., Steinhoff M., Bunnett N. W. Proteinase-activated receptors: novel mechanisms of signaling by serine proteases. Am J Physiol. 1998;274(6 Pt 1):C1429-C1452.

    Not in our library10.1152/ajpcell.1998.274.6.c1429 (opens in a new tab)

16. 16.

    Green G. M., Nasset E. S. Importance of bile in regulation of intraluminal proteolytic enzyme activities in the rat. Gastroenterology. 1980;79(4):695-702.

    Not in our library

17. 17.

    Hedström J., Haglund C., Haapiainen R., Stenman U. H. Serum trypsinogen-2 and trypsin-2-alpha(1)-antitrypsin complex in malignant and benign digestive-tract diseases. Preferential elevation in patients with cholangiocarcinomas. Int J Cancer. 1996;66(3):326-331.

    Not in our library10.1002/(sici)1097-0215(19960503)66:3<326::aid-ijc10>3.0.co;2-9 (opens in a new tab)

18. 18.

    Hirahara F., Miyagi Y., Miyagi E., Yasumitsu H., Koshikawa N., Nagashima Y., Kitamura H., Minaguchi H., Umeda M., Miyazaki K. Trypsinogen expression in human ovarian carcinomas. Int J Cancer. 1995;63(2):176-181.

    Not in our library10.1002/ijc.2910630205 (opens in a new tab)

19. 19.

    Hollenberg M. D. Protease-mediated signalling: new paradigms for cell regulation and drug development. Trends Pharmacol Sci. 1996;17(1):3-6.

    Not in our library10.1016/0165-6147(96)81562-8 (opens in a new tab)

20. 20.

    Hollenberg M. D., Saifeddine M., al-Ani B., Kawabata A. Proteinase-activated receptors: structural requirements for activity, receptor cross-reactivity, and receptor selectivity of receptor-activating peptides. Can J Physiol Pharmacol. 1997;75(7):832-841.

    Not in our library

21. 21.

    Ichikawa Y., Koshikawa N., Hasegawa S., Ishikawa T., Momiyama N., Kunizaki C., Takahashi M., Moriwaki Y., Akiyama H., Yamaoka H. Marked increase of trypsin(ogen) in serum of linitis plastica (gastric cancer, borrmann 4) patients. Clin Cancer Res. 2000;6(4):1385-1388.

    Not in our library

22. 22.

    Ishihara H., Connolly A. J., Zeng D., Kahn M. L., Zheng Y. W., Timmons C., Tram T., Coughlin S. R. Protease-activated receptor 3 is a second thrombin receptor in humans. Nature. 1997;386(6624):502-506.

    Not in our library10.1038/386502a0 (opens in a new tab)

23. 23.

    Kennedy A. R. Prevention of carcinogenesis by protease inhibitors. Cancer Res. 1994;54(7 Suppl):1999s-2005s.

    Not in our library

24. 24.

    Kennedy A. R. The Bowman-Birk inhibitor from soybeans as an anticarcinogenic agent. Am J Clin Nutr. 1998;68(6 Suppl):1406S-1412S.

    Not in our library10.1093/ajcn/68.6.1406s (opens in a new tab)

25. 25.

    Kong W., McConalogue K., Khitin L. M., Hollenberg M. D., Payan D. G., Böhm S. K., Bunnett N. W. Luminal trypsin may regulate enterocytes through proteinase-activated receptor 2. Proc Natl Acad Sci U S A. 1997;94(16):8884-8889.

    Not in our library10.1073/pnas.94.16.8884 (opens in a new tab)

26. 26.

    Koshikawa N., Hasegawa S., Nagashima Y., Mitsuhashi K., Tsubota Y., Miyata S., Miyagi Y., Yasumitsu H., Miyazaki K. Expression of trypsin by epithelial cells of various tissues, leukocytes, and neurons in human and mouse. Am J Pathol. 1998;153(3):937-944.

    Not in our library10.1016/s0002-9440(10)65635-0 (opens in a new tab)

27. 27.

    Koshikawa N., Nagashima Y., Miyagi Y., Mizushima H., Yanoma S., Yasumitsu H., Miyazaki K. Expression of trypsin in vascular endothelial cells. FEBS Lett. 1997;409(3):442-448.

    Not in our library10.1016/s0014-5793(97)00565-6 (opens in a new tab)

28. 28.

    Koshikawa N., Yasumitsu H., Umeda M., Miyazaki K. Multiple secretion of matrix serine proteinases by human gastric carcinoma cell lines. Cancer Res. 1992;52(18):5046-5053.

    Not in our library

29. 29.

    Laburthe M., Rousset M., Rouyer-Fessard C., Couvineau A., Chantret I., Chevalier G., Zweibaum A. Development of vasoactive intestinal peptide-responsive adenylate cyclase during enterocytic differentiation of Caco-2 cells in culture. Evidence for an increased receptor level. J Biol Chem. 1987;262(21):10180-10184.

    Not in our library

30. 30.

    Maoret J. J., Anini Y., Rouyer-Fessard C., Gully D., Laburthe M. Neurotensin and a non-peptide neurotensin receptor antagonist control human colon cancer cell growth in cell culture and in cells xenografted into nude mice. Int J Cancer. 1999;80(3):448-454.

    Not in our library10.1002/(sici)1097-0215(19990129)80:3<448::aid-ijc19>3.0.co;2-n (opens in a new tab)

31. 31.

    Mazzieri R., Masiero L., Zanetta L., Monea S., Onisto M., Garbisa S., Mignatti P. Control of type IV collagenase activity by components of the urokinase-plasmin system: a regulatory mechanism with cell-bound reactants. EMBO J. 1997;16(9):2319-2332.

    Not in our library10.1093/emboj/16.9.2319 (opens in a new tab)

32. 32.

    McCawley L. J., Matrisian L. M. Matrix metalloproteinases: multifunctional contributors to tumor progression. Mol Med Today. 2000;6(4):149-156.

    Not in our library10.1016/s1357-4310(00)01686-5 (opens in a new tab)

33. 33.

    Mignatti P., Rifkin D. B. Biology and biochemistry of proteinases in tumor invasion. Physiol Rev. 1993;73(1):161-195.

    Not in our library10.1152/physrev.1993.73.1.161 (opens in a new tab)

34. 34.

    Mirza H., Schmidt V. A., Derian C. K., Jesty J., Bahou W. F. Mitogenic responses mediated through the proteinase-activated receptor-2 are induced by expressed forms of mast cell alpha- or beta-tryptases. Blood. 1997;90(10):3914-3922.

    Not in our library

35. 35.

    Miyata S., Koshikawa N., Higashi S., Miyagi Y., Nagashima Y., Yanoma S., Kato Y., Yasumitsu H., Miyazaki K. Expression of trypsin in human cancer cell lines and cancer tissues and its tight binding to soluble form of Alzheimer amyloid precursor protein in culture. J Biochem. 1999;125(6):1067-1076.

    Not in our library10.1093/oxfordjournals.jbchem.a022388 (opens in a new tab)

36. 36.

    Miyata S., Miyagi Y., Koshikawa N., Nagashima Y., Kato Y., Yasumitsu H., Hirahara F., Misugi K., Miyazaki K. Stimulation of cellular growth and adhesion to fibronectin and vitronectin in culture and tumorigenicity in nude mice by overexpression of trypsinogen in human gastric cancer cells. Clin Exp Metastasis. 1998;16(7):613-622.

    Not in our library10.1023/a:1006576313979 (opens in a new tab)

37. 37.

    Molino M., Barnathan E. S., Numerof R., Clark J., Dreyer M., Cumashi A., Hoxie J. A., Schechter N., Woolkalis M., Brass L. F. Interactions of mast cell tryptase with thrombin receptors and PAR-2. J Biol Chem. 1997;272(7):4043-4049.

    Not in our library10.1074/jbc.272.7.4043 (opens in a new tab)

38. 38.

    Molino M., Raghunath P. N., Kuo A., Ahuja M., Hoxie J. A., Brass L. F., Barnathan E. S. Differential expression of functional protease-activated receptor-2 (PAR-2) in human vascular smooth muscle cells. Arterioscler Thromb Vasc Biol. 1998;18(5):825-832.

    Not in our library10.1161/01.atv.18.5.825 (opens in a new tab)

39. 39.

    Molino M., Woolkalis M. J., Reavey-Cantwell J., Praticó D., Andrade-Gordon P., Barnathan E. S., Brass L. F. Endothelial cell thrombin receptors and PAR-2. Two protease-activated receptors located in a single cellular environment. J Biol Chem. 1997;272(17):11133-11141.

    Not in our library10.1074/jbc.272.17.11133 (opens in a new tab)

40. 40.

    Nguyen T. D., Moody M. W., Steinhoff M., Okolo C., Koh D. S., Bunnett N. W. Trypsin activates pancreatic duct epithelial cell ion channels through proteinase-activated receptor-2. J Clin Invest. 1999;103(2):261-269.

    Not in our library10.1172/jci2539 (opens in a new tab)

41. 41.

    Normanno N., De Luca A., Salomon D. S., Ciardiello F. Epidermal growth factor-related peptides as targets for experimental therapy of human colon carcinoma. Cancer Detect Prev. 1998;22(1):62-67.

    Not in our library

42. 42.

    Nystedt S., Emilsson K., Larsson A. K., Strömbeck B., Sundelin J. Molecular cloning and functional expression of the gene encoding the human proteinase-activated receptor 2. Eur J Biochem. 1995;232(1):84-89.

    Not in our library10.1111/j.1432-1033.1995.tb20784.x (opens in a new tab)

43. 43.

    Nystedt S., Emilsson K., Wahlestedt C., Sundelin J. Molecular cloning of a potential proteinase activated receptor. Proc Natl Acad Sci U S A. 1994;91(20):9208-9212.

    Not in our library10.1073/pnas.91.20.9208 (opens in a new tab)

44. 44.

    O'Brien P. J., Molino M., Kahn M., Brass L. F. Protease activated receptors: theme and variations. Oncogene. 2001;20(13):1570-1581.

    Not in our library10.1038/sj.onc.1204194 (opens in a new tab)

45. 45.

    Ramos-DeSimone N., Hahn-Dantona E., Sipley J., Nagase H., French D. L., Quigley J. P. Activation of matrix metalloproteinase-9 (MMP-9) via a converging plasmin/stromelysin-1 cascade enhances tumor cell invasion. J Biol Chem. 1999;274(19):13066-13076.

    Not in our library10.1074/jbc.274.19.13066 (opens in a new tab)

46. 46.

    Saifeddine M., al-Ani B., Cheng C. H., Wang L., Hollenberg M. D. Rat proteinase-activated receptor-2 (PAR-2): cDNA sequence and activity of receptor-derived peptides in gastric and vascular tissue. Br J Pharmacol. 1996;118(3):521-530.

    Not in our library10.1111/j.1476-5381.1996.tb15433.x (opens in a new tab)

47. 47.

    Singh P., Rubin N. Insulinlike growth factors and binding proteins in colon cancer. Gastroenterology. 1993;105(4):1218-1237.

    Not in our library10.1016/0016-5085(93)90971-e (opens in a new tab)

48. 48.

    Singh P., Velasco M., Given R., Wargovich M., Varro A., Wang T. C. Mice overexpressing progastrin are predisposed for developing aberrant colonic crypt foci in response to AOM. Am J Physiol Gastrointest Liver Physiol. 2000;278(3):G390-G399.

    Not in our library10.1152/ajpgi.2000.278.3.g390 (opens in a new tab)

49. 49.

    Smith R., Jenkins A., Lourbakos A., Thompson P., Ramakrishnan V., Tomlinson J., Deshpande U., Johnson D. A., Jones R., Mackie E. J. Evidence for the activation of PAR-2 by the sperm protease, acrosin: expression of the receptor on oocytes. FEBS Lett. 2000;484(3):285-290.

    Not in our library10.1016/s0014-5793(00)02146-3 (opens in a new tab)

50. 50.

    Sorsa T., Salo T., Koivunen E., Tyynelä J., Konttinen Y. T., Bergmann U., Tuuttila A., Niemi E., Teronen O., Heikkilä P. Activation of type IV procollagenases by human tumor-associated trypsin-2. J Biol Chem. 1997;272(34):21067-21074.

    Not in our library10.1074/jbc.272.34.21067 (opens in a new tab)

51. 51.

    Vergnolle N., Macnaughton W. K., Al-Ani B., Saifeddine M., Wallace J. L., Hollenberg M. D. Proteinase-activated receptor 2 (PAR2)-activating peptides: identification of a receptor distinct from PAR2 that regulates intestinal transport. Proc Natl Acad Sci U S A. 1998;95(13):7766-7771.

    Not in our library10.1073/pnas.95.13.7766 (opens in a new tab)

52. 52.

    Vergnolle N. Review article: proteinase-activated receptors - novel signals for gastrointestinal pathophysiology. Aliment Pharmacol Ther. 2000;14(3):257-266.

    Not in our library10.1046/j.1365-2036.2000.00690.x (opens in a new tab)

53. 53.

    Vu T. K., Hung D. T., Wheaton V. I., Coughlin S. R. Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell. 1991;64(6):1057-1068.

    Not in our library10.1016/0092-8674(91)90261-v (opens in a new tab)

54. 54.

    Xu W. F., Andersen H., Whitmore T. E., Presnell S. R., Yee D. P., Ching A., Gilbert T., Davie E. W., Foster D. C. Cloning and characterization of human protease-activated receptor 4. Proc Natl Acad Sci U S A. 1998;95(12):6642-6646.

    Not in our library10.1073/pnas.95.12.6642 (opens in a new tab)

55. 55.

    Yu Z., Ahmad S., Schwartz J. L., Banville D., Shen S. H. Protein-tyrosine phosphatase SHP2 is positively linked to proteinase-activated receptor 2-mediated mitogenic pathway. J Biol Chem. 1997;272(11):7519-7524.

    Not in our library10.1074/jbc.272.11.7519 (opens in a new tab)

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