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

Abstract

Summary 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 α 1 -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. 1363-1371 © 2001 Cancer Research Campaign doi: 10.1054/ bjoc.2001.1806, available online at http://www.idealibrary.com on http://www.bjcancer.com aminoterminal domain for the expression of activity (Tryggvason et al, 1987). The urokinase type plasminogen activator (uPA) -plasmin system (Mazzieri et al, 1997) and membrane type MMP-1 (MT1-MMP, MMP-14) (Sato et al, 1994;Tokuraku et al, 1995) have been suggested to represent physiological mechanisms for the control of MMP-2 and MMP-9 activity. Recent studies have shown that human trypsin-2 is more potent than plasmin and other serine proteinases in activating several MMPs, including MMP-2 and MMP-9 (Sorsa et al, 1997), and that downregulation of trypsin-2 expression and activity in colon cancer cells is associated with decreased proMMP-9 activation (Lukkonen et al, 2000).In the present study we investigated the involvement of trypsin in the activation of MMP-2 and MMP-9 in vivo by studying whether the concentrations of trypsins, trypsin-API-complexes, and TATI are associated with activation of proMMP-2 and proMMP-9 in ovarian tumour cyst fluid. Furthermore, we studied the immunohistochemical expression and localization of trypsinogen-2, TATI, MMP-2, and MMP-9 in ovarian tumours. MATERIALS AND METHODS Clinical specimensCyst fluid samples (n = 61) were obtained from 58 patients (age range 13-79 years) undergoing surgery for removal of the tumour during 1986-1998 at Helsinki University Central Hospital. Of the tumours, 41 were benign...

Expression of Trypsinogen-1, Trypsinogen-2, and Tumor-Associated Trypsin Inhibitor in Ovarian Cancer

“…Trypsin-1 and trypsin-2 are thought to be involved in the spread of ovarian cancer by degrading extracellular matrix (12) and by activating other proteases associated with cancer invasion, e.g., urokinase-type plasminogen activator and MMPs (13,15,46). Trypsinogen (17,18) and TATI (27,47) are often expressed in ovarian cancer, and immunohistochemical expression of trypsinogen-1 has been reported to be more frequent in malignant than in benign or borderline tumors (19).…”

“…Trypsinogen (17,18) and TATI (27,47) are often expressed in ovarian cancer, and immunohistochemical expression of trypsinogen-1 has been reported to be more frequent in malignant than in benign or borderline tumors (19). Trypsinogen-1, trypsinogen-2, and TATI occur at high concentrations in cyst fluid produced by ovarian tumors (15,17). These, as well as complexes of API with trypsin-1 and trypsin-2, which reflect trypsin activation, occur at higher concentrations in cyst fluid from malignant than from benign ovarian tumors (15,17), and this is associated with MMP-9 activation (15).…”

“…Trypsinogen-1, trypsinogen-2, and TATI occur at high concentrations in cyst fluid produced by ovarian tumors (15,17). These, as well as complexes of API with trypsin-1 and trypsin-2, which reflect trypsin activation, occur at higher concentrations in cyst fluid from malignant than from benign ovarian tumors (15,17), and this is associated with MMP-9 activation (15).…”

“…It degrades a wide spectrum of extracellular matrix proteins (12) and activates proforms of other proteinases, including MMPs 1, 2, 8, 9, and 13 (13)(14)(15)(16). Tumor-associated trypsinogen-1 and trypsinogen-2 are strongly expressed in most ovarian cancers (17)(18)(19) and in several other cancers, e.g., pancreatic (20), gastric (21,22) and colorectal cancer (23,24), cholangiocarcinoma (25), and in esophageal squamous cell carcinoma (26).…”

Role of the tumor-associated trypsin inhibitor SPINK1 in cancer development

“…5 This notion was supported by the finding of high tissue expression of trypsinogen-2 and adverse prognosis in ovarian cancer. 8 In other tumors, low expression of SPINK1 is associated with adverse prognosis and aggressive disease, i.e., ventricular and bladder cancer. 9,10 In these tumors, SPINK1 may inhibit tumor invasion by inhibiting protease cascades involving trypsin.…”