Human colon carcinoma, fibrosarcoma and leukemia cell lines produce tumor‐associated trypsinogen

Abstract

Previous studies have indicated that cyst fluid of ovarian tumors contains 2 trypsinogen isoenzymes, called tumor-associated trypsinogen-I (TAT-I) and trypsinogen-2 (TAT-2), the levels of which correlate with the degree of malignancy of the tumors. In addition, these cyst fluids contain large amounts of tumor-associated trypsin inhibitor (TATI), which is also expressed in many other human tumors. In the present study we examined the production of TAT-I, TAT-2 and TATI in 9 established tumor-cell lines. TAT-2 was produced by 5 cell lines. Its concentration in the conditioned medium of COLO 205 colon adenocarcinoma cells, K-562 erythroleukemia cells and fibrosarcoma cell lines HT 1080, 8387 and A 9733 was 460 micrograms/l, 9.8 micrograms/l, 21 micrograms/l, 8.8 micrograms/l and 0.24 micrograms/l, respectively. TAT-I was detectable in the conditioned medium of COLO 205 and HT 1080 cells at concentrations of 64 micrograms/l and 0.5 micrograms/l, respectively. TATI was detected only in the media of COLO 205 cells at a concentration of 23 micrograms/l. TAT-2 zymogen was purified from the conditioned medium of COLO 205 and HT 1080 cells by immunoaffinity chromatography. According to its aminoterminal amino acid sequence, a molecular mass of 28 kDa by SDS-PAGE, elution pattern in ion-exchange chromatography and ability to be activated by enteropeptidase, the zymogen is identical to that previously isolated from cyst fluid of ovarian tumors. In addition, we found that TAT-2 secretion could be down-regulated by dexamethasone in HT 1080 cells but not in COLO 205 cells. The abundant production of TAT-2 isoenzyme in different cancer cell lines suggests that it could contribute to the increased proteolytic activity of many human tumors.

Molecular cloning, expression and potential functions of the human proteinase-activated receptor-2

“…Elsewhere, potential roles of PAR-2 are less obvious. However, trypsin-like enzymes and pancreatic secretory trypsin inhibitors are present in non-pancreatic tissues, tumours and tumour cell lines, where PAR-2 may regulate growth [19][20][21][22].…”

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

“…Concentration (µg/l) P < 0.005 P = 0.01 P < 0.0001 1991b;Koshikawa et al, 1994). The main tumour-associated isoenzyme, trypsin-2, activates pro-urokinase-type plasminogen activator (pro-uPA) (Koivunen et al, 1989) and in vitro it is the most efficient activator of the 92 kDa gelatinase B (MMP-9) known thus far (Sorsa et al, 1997).…”

“…Recently, extrapancreatic expression of trypsinogen has been observed in several human malignancies such as ovarian (Koivunen et al, 1989;Hirahara et al, 1995) and gastric cancer (Fujimura et al, 1998) and in cholangiocarcinoma (Terada et al, 1995). Several tumour cell lines also express trypsinogen (Koivunen et al, 1991b;Koshikawa et al, 1994). In human ovarian and gastric cancer, the trypsin expression is associated with the malignant potential of the tumours Hirahara et al, 1998;Miyata et al, 1998;Kato et al, 1998).…”

Epidermal growth factor‐binding protein activates soluble and receptor‐bound single chain urokinase‐type plasminogen activator

“…By contrast, EGF-BP demonstrated a 2-fold greater catalytic efficiency than the related proteinase NGF-7 (Table 1) and over a 10-fold greater catalytic efficiency than mast cell tryptase (2.4x 103 M-l.s -1, [9]). Comparison of EGF-BP with other scu-PA activating enzymes such as plasma kallikrein, tumorassociated trypsin, and cathepsins B and L, is difficult since formal kinetic parameters (kc,t, K~) have not been determined for these enzymes [4][5][6][7]. Since all these proteinases activate scu-PA less efficiently than plasmin, they most likely function in the initiation of the u-PA-dependent cascade and not in the amplification phase.…”

“…Since plasmin is present in biological fluids as an inactive zymogen (plasminogen), a critical question remains regarding initiation of u-PA-dependent cell surface proteolysis. Several proteinases other than plasmin activate scu-PA, including plasma kallikrein, tumor-associated trypsin, cathepsin B, cathepsin L, nerve growth factor-), (NGF-7), mast cell tryptase, and human T cell-associated serine proteinase [4][5][6][7][8][9][10]. Although these proteinases activate scu-PA less efficiently than plasmin, they may initiate the u-PA dependent proteinase cascade which can then undergo self-amplification.…”