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. 2001 May;84(10):1363–1371. doi: 10.1054/bjoc.2001.1806

The levels of trypsinogen isoenzymes in ovarian tumour cyst fluids are associated with promatrix metalloproteinase-9 but not promatrix metalloproteinase-2 activation

A Paju 1, T Sorsa 2, T Tervahartiala 2, E Koivunen 3, C Haglund 4, A Leminen 5, T Wahlström 6, T Salo 7, U-H Stenman 1
PMCID: PMC2363633  PMID: 11355948

Abstract

Proteolysis mediated by matrix metalloproteinases (MMPs) and serine proteinases is associated with cancer invasion and metastasis. Activation of latent proMMPs, and especially the proforms of the type IV collagen degrading gelatinases A and B (proMMP-2 and proMMP-9), is thought to be a critical step in this process. We have recently found that human tumour-associated trypsin-2 is a potent activator of proMMP-9 and it also activates proMMP-2 in vitro. Trypsinogen, MMP-2, and MMP-9 are expressed in ovarian cancer. To elucidate the function of trypsin in vivo, we studied whether high concentrations of trypsinogen-1, trypsinogen-2, their α1-proteinase inhibitor (API) complexes, and tumour-associated trypsin inhibitor (TATI) are associated with proMMP-2 and proMMP-9 activation in ovarian tumour cyst fluids. Zymography and immunofluorometric analysis of 61 cyst fluids showed a significant association between high trypsin concentrations and the activation of MMP-9 (P= 0.003–0.05). In contrast, the trypsin concentrations were inversely associated with the activation of MMP-2 (P= 0.01–0.02). Immunohistochemical analysis of ovarian tumour tissue demonstrated expression of trypsinogen-2 and TATI in the secretory epithelium. MMP-2 was detected both in stromal and epithelial cells whereas MMP-9 was detected in neutrophils and macrophage-like cells in stromal and epithelial areas. These results suggest that trypsin may play a role in the regulation of the MMP-dependent proteolysis associated with invasion and metastasis of ovarian cancer. © 2001 Cancer Research Campaign www.bjcancer.com

Keywords: trypsin, TATI, MMP-2, MMP-9, ovarian cancer, cyst fluid

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Selected References

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  1. Afzal S., Lalani el-N, Foulkes W. D., Boyce B., Tickle S., Cardillo M. R., Baker T., Pignatelli M., Stamp G. W. Matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expression and synthetic matrix metalloproteinase-2 inhibitor binding in ovarian carcinomas and tumor cell lines. Lab Invest. 1996 Feb;74(2):406–421. [PubMed] [Google Scholar]
  2. Afzal S., Lalani E. N., Poulsom R., Stubbs A., Rowlinson G., Sato H., Seiki M., Stamp G. W. MT1-MMP and MMP-2 mRNA expression in human ovarian tumors: possible implications for the role of desmoplastic fibroblasts. Hum Pathol. 1998 Feb;29(2):155–165. doi: 10.1016/s0046-8177(98)90226-x. [DOI] [PubMed] [Google Scholar]
  3. Autio-Harmainen H., Karttunen T., Hurskainen T., Höyhtyä M., Kauppila A., Tryggvason K. Expression of 72 kilodalton type IV collagenase (gelatinase A) in benign and malignant ovarian tumors. Lab Invest. 1993 Sep;69(3):312–321. [PubMed] [Google Scholar]
  4. Benedet J. L., Bender H., Jones H., 3rd, Ngan H. Y., Pecorelli S. FIGO staging classifications and clinical practice guidelines in the management of gynecologic cancers. FIGO Committee on Gynecologic Oncology. Int J Gynaecol Obstet. 2000 Aug;70(2):209–262. [PubMed] [Google Scholar]
  5. Bernhard E. J., Gruber S. B., Muschel R. J. Direct evidence linking expression of matrix metalloproteinase 9 (92-kDa gelatinase/collagenase) to the metastatic phenotype in transformed rat embryo cells. Proc Natl Acad Sci U S A. 1994 May 10;91(10):4293–4297. doi: 10.1073/pnas.91.10.4293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boyd R. S., Balkwill F. R. MMP-2 release and activation in ovarian carcinoma: the role of fibroblasts. Br J Cancer. 1999 May;80(3-4):315–321. doi: 10.1038/sj.bjc.6690357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Davies B., Miles D. W., Happerfield L. C., Naylor M. S., Bobrow L. G., Rubens R. D., Balkwill F. R. Activity of type IV collagenases in benign and malignant breast disease. Br J Cancer. 1993 May;67(5):1126–1131. doi: 10.1038/bjc.1993.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davies B., Waxman J., Wasan H., Abel P., Williams G., Krausz T., Neal D., Thomas D., Hanby A., Balkwill F. Levels of matrix metalloproteases in bladder cancer correlate with tumor grade and invasion. Cancer Res. 1993 Nov 15;53(22):5365–5369. [PubMed] [Google Scholar]
  9. De Nictolis M., Garbisa S., Lucarini G., Goteri G., Masiero L., Ciavattini A., Garzetti G. G., Stetler-Stevenson W. G., Fabris G., Biagini G. 72-kilodalton type IV collagenase, type IV collagen, and Ki 67 antigen in serous tumors or the ovary: a clinicopathologic, immunohistochemical, and Serological study. Int J Gynecol Pathol. 1996 Apr;15(2):102–109. doi: 10.1097/00004347-199604000-00003. [DOI] [PubMed] [Google Scholar]
  10. Ferry G., Lonchampt M., Pennel L., de Nanteuil G., Canet E., Tucker G. C. Activation of MMP-9 by neutrophil elastase in an in vivo model of acute lung injury. FEBS Lett. 1997 Feb 3;402(2-3):111–115. doi: 10.1016/s0014-5793(96)01508-6. [DOI] [PubMed] [Google Scholar]
  11. Fishman D. A., Bafetti L. M., Banionis S., Kearns A. S., Chilukuri K., Stack M. S. Production of extracellular matrix-degrading proteinases by primary cultures of human epithelial ovarian carcinoma cells. Cancer. 1997 Oct 15;80(8):1457–1463. doi: 10.1002/(sici)1097-0142(19971015)80:8<1457::aid-cncr13>3.0.co;2-4. [DOI] [PubMed] [Google Scholar]
  12. Fujimura T., Ohta T., Kitagawa H., Fushida S., Nishimura G. I., Yonemura Y., Elnemr A., Miwa K., Nakanuma Y. Trypsinogen expression and early detection for peritoneal dissemination in gastric cancer. J Surg Oncol. 1998 Oct;69(2):71–75. doi: 10.1002/(sici)1096-9098(199810)69:2<71::aid-jso5>3.0.co;2-r. [DOI] [PubMed] [Google Scholar]
  13. Furuya M., Ishikura H., Kawarada Y., Ogawa Y., Sakuragi N., Fujimoto S., Yoshiki T. Expression of matrix metalloproteinases and related tissue inhibitors in the cyst fluids of ovarian mucinous neoplasms. Gynecol Oncol. 2000 Aug;78(2):106–112. doi: 10.1006/gyno.2000.5856. [DOI] [PubMed] [Google Scholar]
  14. Garzetti G. G., Ciavattini A., Lucarini G., Goteri G., de e Nictolis M., Garbisa S., Masiero L., Romanini C., Graziella B. Tissue and serum metalloproteinase (MMP-2) expression in advanced ovarian serous cystoadenocarcinomas: clinical and prognostic implications. Anticancer Res. 1995 Nov-Dec;15(6B):2799–2804. [PubMed] [Google Scholar]
  15. Hedström J., Haglund C., Kemppainen E., Leinimaa M., Leinonen J., Stenman U. H. Time-resolved immunofluorometric assay of trypsin-1 complexed with alpha(1)-antitrypsin in serum: increased immunoreactivity in patients with biliary tract cancer. Clin Chem. 1999 Oct;45(10):1768–1773. [PubMed] [Google Scholar]
  16. Hedström J., Leinonen J., Sainio V., Stenman U. H. Time-resolved immunofluorometric assay of trypsin-2 complexed with alpha 1-antitrypsin in serum. Clin Chem. 1994 Sep;40(9):1761–1765. [PubMed] [Google Scholar]
  17. Hirahara F., Miyagi E., Nagashima Y., Miyagi Y., Yasumitsu H., Koshikawa N., Nakatani Y., Nakazawa T., Udagawa K., Kitamura H. Differential expression of trypsin in human ovarian carcinomas and low-malignant-potential tumors. Gynecol Oncol. 1998 Feb;68(2):162–165. doi: 10.1006/gyno.1997.4912. [DOI] [PubMed] [Google Scholar]
  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 Oct 9;63(2):176–181. doi: 10.1002/ijc.2910630205. [DOI] [PubMed] [Google Scholar]
  19. Huang L. W., Garrett A. P., Bell D. A., Welch W. R., Berkowitz R. S., Mok S. C. Differential expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 protein and mRNA in epithelial ovarian tumors. Gynecol Oncol. 2000 Jun;77(3):369–376. doi: 10.1006/gyno.2000.5806. [DOI] [PubMed] [Google Scholar]
  20. Höyhtyä M., Fridman R., Komarek D., Porter-Jordan K., Stetler-Stevenson W. G., Liotta L. A., Liang C. M. Immunohistochemical localization of matrix metalloproteinase 2 and its specific inhibitor TIMP-2 in neoplastic tissues with monoclonal antibodies. Int J Cancer. 1994 Feb 15;56(4):500–505. doi: 10.1002/ijc.2910560408. [DOI] [PubMed] [Google Scholar]
  21. Höyhtyä M., Hujanen E., Turpeenniemi-Hujanen T., Thorgeirsson U., Liotta L. A., Tryggvason K. Modulation of type-IV collagenase activity and invasive behavior of metastatic human melanoma (A2058) cells in vitro by monoclonal antibodies to type-IV collagenase. Int J Cancer. 1990 Aug 15;46(2):282–286. doi: 10.1002/ijc.2910460224. [DOI] [PubMed] [Google Scholar]
  22. Itkonen O., Koivunen E., Hurme M., Alfthan H., Schröder T., Stenman U. H. Time-resolved immunofluorometric assays for trypsinogen-1 and 2 in serum reveal preferential elevation of trypsinogen-2 in pancreatitis. J Lab Clin Med. 1990 Jun;115(6):712–718. [PubMed] [Google Scholar]
  23. Juarez J., Clayman G., Nakajima M., Tanabe K. K., Saya H., Nicolson G. L., Boyd D. Role and regulation of expression of 92-kDa type-IV collagenase (MMP-9) in 2 invasive squamous-cell-carcinoma cell lines of the oral cavity. Int J Cancer. 1993 Aug 19;55(1):10–18. doi: 10.1002/ijc.2910550104. [DOI] [PubMed] [Google Scholar]
  24. Kato Y., Nagashima Y., Koshikawa N., Miyagi Y., Yasumitsu H., Miyazaki K. Production of trypsins by human gastric cancer cells correlates with their malignant phenotype. Eur J Cancer. 1998 Jun;34(7):1117–1123. doi: 10.1016/s0959-8049(98)00077-x. [DOI] [PubMed] [Google Scholar]
  25. Kjeldsen L., Johnsen A. H., Sengeløv H., Borregaard N. Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. J Biol Chem. 1993 May 15;268(14):10425–10432. [PubMed] [Google Scholar]
  26. Koivunen E., Huhtala M. L., Stenman U. H. Human ovarian tumor-associated trypsin. Its purification and characterization from mucinous cyst fluid and identification as an activator of pro-urokinase. J Biol Chem. 1989 Aug 25;264(24):14095–14099. [PubMed] [Google Scholar]
  27. Koivunen E., Itkonen O., Halila H., Stenman U. H. Cyst fluid of ovarian cancer patients contains high concentrations of trypsinogen-2. Cancer Res. 1990 Apr 15;50(8):2375–2378. [PubMed] [Google Scholar]
  28. Koivunen E., Ristimäki A., Itkonen O., Osman S., Vuento M., Stenman U. H. Tumor-associated trypsin participates in cancer cell-mediated degradation of extracellular matrix. Cancer Res. 1991 Apr 15;51(8):2107–2112. [PubMed] [Google Scholar]
  29. Koivunen E., Saksela O., Itkonen O., Osman S., Huhtala M. L., Stenman U. H. Human colon carcinoma, fibrosarcoma and leukemia cell lines produce tumor-associated trypsinogen. Int J Cancer. 1991 Feb 20;47(4):592–596. doi: 10.1002/ijc.2910470419. [DOI] [PubMed] [Google Scholar]
  30. Koshikawa N., Yasumitsu H., Nagashima Y., Umeda M., Miyazaki K. Identification of one- and two-chain forms of trypsinogen 1 produced by a human gastric adenocarcinoma cell line. Biochem J. 1994 Oct 1;303(Pt 1):187–190. doi: 10.1042/bj3030187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  32. Levy A. T., Cioce V., Sobel M. E., Garbisa S., Grigioni W. F., Liotta L. A., Stetler-Stevenson W. G. Increased expression of the Mr 72,000 type IV collagenase in human colonic adenocarcinoma. Cancer Res. 1991 Jan 1;51(1):439–444. [PubMed] [Google Scholar]
  33. Liotta L. A., Tryggvason K., Garbisa S., Hart I., Foltz C. M., Shafie S. Metastatic potential correlates with enzymatic degradation of basement membrane collagen. Nature. 1980 Mar 6;284(5751):67–68. doi: 10.1038/284067a0. [DOI] [PubMed] [Google Scholar]
  34. Lukkonen A., Sorsa T., Salo T., Tervahartiala T., Koivunen E., Golub L., Simon S., Stenman U. H. Down-regulation of trypsinogen-2 expression by chemically modified tetracyclines: association with reduced cancer cell migration. Int J Cancer. 2000 May 15;86(4):577–581. doi: 10.1002/(sici)1097-0215(20000515)86:4<577::aid-ijc21>3.0.co;2-j. [DOI] [PubMed] [Google Scholar]
  35. 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 May 1;16(9):2319–2332. doi: 10.1093/emboj/16.9.2319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Mignatti P., Robbins E., Rifkin D. B. Tumor invasion through the human amniotic membrane: requirement for a proteinase cascade. Cell. 1986 Nov 21;47(4):487–498. doi: 10.1016/0092-8674(86)90613-6. [DOI] [PubMed] [Google Scholar]
  37. 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 Oct;16(7):613–622. doi: 10.1023/a:1006576313979. [DOI] [PubMed] [Google Scholar]
  38. Moser T. L., Young T. N., Rodriguez G. C., Pizzo S. V., Bast R. C., Jr, Stack M. S. Secretion of extracellular matrix-degrading proteinases is increased in epithelial ovarian carcinoma. Int J Cancer. 1994 Feb 15;56(4):552–559. doi: 10.1002/ijc.2910560415. [DOI] [PubMed] [Google Scholar]
  39. Naylor M. S., Stamp G. W., Davies B. D., Balkwill F. R. Expression and activity of MMPS and their regulators in ovarian cancer. Int J Cancer. 1994 Jul 1;58(1):50–56. doi: 10.1002/ijc.2910580110. [DOI] [PubMed] [Google Scholar]
  40. Nielsen B. S., Timshel S., Kjeldsen L., Sehested M., Pyke C., Borregaard N., Danø K. 92 kDa type IV collagenase (MMP-9) is expressed in neutrophils and macrophages but not in malignant epithelial cells in human colon cancer. Int J Cancer. 1996 Jan 3;65(1):57–62. doi: 10.1002/(SICI)1097-0215(19960103)65:1<57::AID-IJC10>3.0.CO;2-F. [DOI] [PubMed] [Google Scholar]
  41. Ohlsson K. Acute pancreatitis. Biochemical, pathophysiological and therapeutics aspects. Acta Gastroenterol Belg. 1988 Jan-Feb;51(1):3–12. [PubMed] [Google Scholar]
  42. Osman S., Turpeinen U., Itkonen O., Stenman U. H. Optimization of a time-resolved immunofluorometric assay for tumor-associated trypsin inhibitor (TATI) using the streptavidin-biotin system. J Immunol Methods. 1993 May 5;161(1):97–106. doi: 10.1016/0022-1759(93)90201-h. [DOI] [PubMed] [Google Scholar]
  43. Pyke C., Ralfkiaer E., Tryggvason K., Danø K. Messenger RNA for two type IV collagenases is located in stromal cells in human colon cancer. Am J Pathol. 1993 Feb;142(2):359–365. [PMC free article] [PubMed] [Google Scholar]
  44. Sato H., Takino T., Okada Y., Cao J., Shinagawa A., Yamamoto E., Seiki M. A matrix metalloproteinase expressed on the surface of invasive tumour cells. Nature. 1994 Jul 7;370(6484):61–65. doi: 10.1038/370061a0. [DOI] [PubMed] [Google Scholar]
  45. 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 Aug 22;272(34):21067–21074. doi: 10.1074/jbc.272.34.21067. [DOI] [PubMed] [Google Scholar]
  46. Stenman U. H., Koivunen E., Itkonen O. Biology and function of tumor-associated trypsin inhibitor, TATI. Scand J Clin Lab Invest Suppl. 1991;207:5–9. doi: 10.3109/00365519109104618. [DOI] [PubMed] [Google Scholar]
  47. Terada T., Ohta T., Minato H., Nakanuma Y. Expression of pancreatic trypsinogen/trypsin and cathepsin B in human cholangiocarcinomas and hepatocellular carcinomas. Hum Pathol. 1995 Jul;26(7):746–752. doi: 10.1016/0046-8177(95)90222-8. [DOI] [PubMed] [Google Scholar]
  48. Tokuraku M., Sato H., Murakami S., Okada Y., Watanabe Y., Seiki M. Activation of the precursor of gelatinase A/72 kDa type IV collagenase/MMP-2 in lung carcinomas correlates with the expression of membrane-type matrix metalloproteinase (MT-MMP) and with lymph node metastasis. Int J Cancer. 1995 Oct 20;64(5):355–359. doi: 10.1002/ijc.2910640513. [DOI] [PubMed] [Google Scholar]
  49. Tryggvason K., Höyhtyä M., Salo T. Proteolytic degradation of extracellular matrix in tumor invasion. Biochim Biophys Acta. 1987 Nov 25;907(3):191–217. doi: 10.1016/0304-419x(87)90006-0. [DOI] [PubMed] [Google Scholar]
  50. Turpeenniemi-Hujanen T., Rönnberg L., Kauppila A., Puistola U. Laminin in the human embryo implantation: analogy to the invasion by malignant cells. Fertil Steril. 1992 Jul;58(1):105–113. doi: 10.1016/s0015-0282(16)55145-0. [DOI] [PubMed] [Google Scholar]
  51. Ueda G., Shimizu C., Tanaka Y., Inoue M., Tanizawa O., Ogawa M., Mori T. Immunohistochemical demonstration of pancreatic secretory trypsin inhibitor in gynecologic tumors. Gynecol Oncol. 1989 Jan;32(1):37–40. doi: 10.1016/0090-8258(89)90846-9. [DOI] [PubMed] [Google Scholar]
  52. Venesmaa P., Stenman U. H., Forss M., Leminen A., Lehtovirta P., Vartiainen J., Paavonen J. Pre-operative serum level of tumour-associated trypsin inhibitor and residual tumour size as prognostic indicators in Stage III epithelial ovarian cancer. Br J Obstet Gynaecol. 1998 May;105(5):508–511. doi: 10.1111/j.1471-0528.1998.tb10150.x. [DOI] [PubMed] [Google Scholar]
  53. Vergote I. B., Kaern J., Abeler V. M., Pettersen E. O., De Vos L. N., Tropé C. G. Analysis of prognostic factors in stage I epithelial ovarian carcinoma: importance of degree of differentiation and deoxyribonucleic acid ploidy in predicting relapse. Am J Obstet Gynecol. 1993 Jul;169(1):40–52. doi: 10.1016/0002-9378(93)90129-7. [DOI] [PubMed] [Google Scholar]
  54. Väisänen A., Kallioinen M., Taskinen P. J., Turpeenniemi-Hujanen T. Prognostic value of MMP-2 immunoreactive protein (72 kD type IV collagenase) in primary skin melanoma. J Pathol. 1998 Sep;186(1):51–58. doi: 10.1002/(SICI)1096-9896(199809)186:1<51::AID-PATH131>3.0.CO;2-P. [DOI] [PubMed] [Google Scholar]
  55. Westerlund A., Hujanen E., Puistola U., Turpeenniemi-Hujanen T. Fibroblasts stimulate human ovarian cancer cell invasion and expression of 72-kDa gelatinase A (MMP-2). Gynecol Oncol. 1997 Oct;67(1):76–82. doi: 10.1006/gyno.1997.4808. [DOI] [PubMed] [Google Scholar]

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