trypsinogen and Carcinoma

There are more than 100 distinct types of cancer, and subtypes can be found within specific organs. Cancer progression is a complex multi-step process. These steps reflect alterations that drive the progressive transformation of normal cells into highly malignant ones. One critical step in tumor growth and invasion is the proteolytic processing of the extracellular matrix environment. The degradation of the extracellular matrix not only enables cell migration, invasion, and metastasis formation, but also affects cell behavior in multiple ways; on one hand by cleaving extracellular matrix bound growth factors and on the other hand by inhibiting angiogenesis into the tumor by liberating cryptic endogenous inhibitors of angiogenesis. Serine proteases and matrix metalloproteases are families of proteolytic enzymes involved in physiological and pathological extracellular matrix and basement membrane processing. In this review, we will focus on the role and activation of trypsinogens, a family of serine proteases, in cancer progression.

Topics: Angiogenesis Inhibitors; Animals; Carcinoma; Cell Transformation, Neoplastic; Extracellular Matrix; Humans; Matrix Metalloproteinases; Neoplasm Invasiveness; Neoplasms; Neovascularization, Pathologic; Trypsin; Trypsinogen

Tumor-associated trypsin-2 and matrix metalloprotease-9 (MMP-9) are associated with cancer, particularly with invasive squamous cell carcinomas. They require activation for catalytical competence via proteolytic cascades. One cascade is formed by enterokinase, trypsin-2 and MMP-9; enterokinase activates trypsinogen-2 to trypsin-2, which is an efficient proMMP-9 activator. We describe here that oral squamous cell carcinomas express all members of this cascade: MMP-9, trypsin-2 and enterokinase. The expression of enterokinase in a carcinoma cell line not derived from the duodenum was shown here for the first time. Enterokinase directly cleaved proMMP-9 at the Lys65-Ser66 site, but failed to activate it in vitro. We demonstrated by confocal microscopy that MMP-9 and trypsin-2 co-localized in intracellular vesicles of the carcinoma cells. This co-localization of trypsin-2 and MMP-9 resulted in intracellular proMMP-9 processing that represented fully or partially activated MMP-9. However, although both proteases were present also in various bone tumor tissues, MMP-9 and trypsin-2 never co-localized at the cellular level in these tissues. This suggests that the intracellular vesicular co-localization, storage and possible activation of these proteases may be a unique feature for aggressive epithelial tumors, such as squamous cell carcinomas, but not for tumors of mesenchymal origin.

Topics: Bone Neoplasms; Carcinoma; Carcinoma, Squamous Cell; Cell Line, Tumor; Enteropeptidase; Enzyme Precursors; Humans; Matrix Metalloproteinase 9; Mouth Neoplasms; Trypsin; Trypsinogen

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.

Topics: Carcinoma; Colonic Neoplasms; Dexamethasone; Extracellular Matrix; Fibrosarcoma; Humans; Isoenzymes; Leukemia; Trypsinogen; Tumor Cells, Cultured

Treatment of AR4-2J cells with dexamethasone at 10 nM for 96 h inhibited cell replication by 75% and increased cell size (30%), protein content (1.6-fold) and protein synthesis (2-fold). The increase in protein synthesis was largely due to a 5 to 10-fold increase in the synthesis of secretory proteins. Amylase activity increased 20 to 30-fold in cellular homogenates and 10 to 20-fold in culture medium. Both in the presence and absence of dexamethasone AR4-2J cells release their secretory proteins by constitutive secretion. The proportion of newly synthesized amylase retained by the cells over the 14 h labeling period increased from 15 to 30% with hormone treatment. As judged by comigration on polyacrylamide gels and Western blots analyzed by immunospecific sera, AR4-2J cells synthesize and secrete the majority of known pancreatic secretory proteins. Dexamethasone increased the synthesis of trypsinogen 12 to 16-fold, chymotrypsinogen 4.5 to 6-fold, the group of procarboxypeptidases 6-fold, and amylase 7 to 10-fold. Messenger RNA levels for trypsinogen, amylase and lipase were each increased 4 to 5-fold. At the ultrastructural level dexamethasone led to significant increases in rough endoplasmic reticulum (RER) (30-fold) and Golgi elements (1.5-fold) and to the de novo appearance of electron-opaque granules (0.1-0.5 microns) which were shown to contain amylase by immunolocalization techniques employing protein A-gold. Dexamethasone also led to the formation of gap junctions between AR4-2J cells. These findings indicate that AR4-2J cells provide a model for differentiation of pancreatic acinar cells which should also be studied for the differentiation markers for the regulated secretory pathway.

Topics: Amylases; Animals; Carboxypeptidases; Carcinoma; Cell Line; Chymotrypsinogen; Cytoplasmic Granules; Endoplasmic Reticulum; Glucocorticoids; Golgi Apparatus; Microscopy, Electron; Neoplasm Proteins; Pancreatic Neoplasms; Rats; Trypsinogen; Tumor Cells, Cultured

Immunoreactive trypsinogen and pancreatic secretory trypsin inhibitor (PSTI) were demonstrated in pancreas by means of an immunoperoxidase technique. They had the same distribution in acinar cells of 'normal' human exocrine pancreas tissues. Ductal adenocarcinoma tissue and pancreatic undifferentiated carcinoma contained neither antigen. Scattered 'normal'-looking cells in the border area between normal and neoplastic tissue of both types of tumor stained positively for trypsinogen and for PSTI.

Topics: Adenocarcinoma; Carcinoma; Humans; Immunoenzyme Techniques; Pancreas; Pancreatic Neoplasms; Trypsin Inhibitor, Kazal Pancreatic; Trypsin Inhibitors; Trypsinogen

Pancreatic carcinomas electively induced by immunological mechanism [(1), (2)] keep their exocrine secretory specificity along the various stages of their evolution: (a) during the transformation phase from adenoma to carcinoma; (b) in the evolved carcinoma; (c) in its metastasis; (d) in the ascitic carcinomatous cells formed. They are called: immuno-inducted carcinoma. The carcinomatous cells of the constantly deadly ascites cease their production of secretion granules after passages by intraperitoneal graft; but this secretion reappears in the solid carcinomas they induce by subcutaneous graft and contains trypsinogen and chymotrypsinogen, even after the 92nd passage, at the 757 day. Besides, the antisera (1) enhance the growth and the affinity for pancreas and adipose tissue of the carcinomatous ascitic strains they induced. They, sometimes, produce nodular hepatic carcinomas.

Topics: Antibodies; Ascites; Carcinoma; Chymotrypsinogen; Lipoprotein Lipase; Lymph Nodes; Lymphatic Metastasis; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Pancreatic Neoplasms; Trypsinogen