Metastatic colorectal cancer stimulates collagen synthesis by fibroblasts.

BACKGROUND: We ascertained in vitro whether there were any differences in the growth of fibroblasts and the production of collagen (PIIIP), in relation to the presence of conditioning sera or tumor tissues from colorectal cancer (CRC) patients with disease progression or regression after surgery. PATIENTS AND METHODS: Fibroblasts were conditioned with: 1) 10% of control (n=20) or CRC patients' (n=57) sera; 2) 10% of tumor tissue homogenates obtained from CRC patients without (group A, n=6) or with (group B, n=5) liver metastases. After surgery, 29/57 patients (group 1) developed while 28/57 (group 2) did not develop a recurrent disease. RESULTS: The ratio between cell growth and PIIIP production increases when fibroblasts were conditioned by group I (p<0.05), but not by controls or group 2 sera. Tumor homogenates from group B inhibited cell growth (p<0.001), while they induced PIIIP production (p<0.001) in comparison with results from group A. CONCLUSION: CRC non-metastatic cancer cells induce mainly fibroblasts growth, while metastatic cells mainly induce collagen synthesis. This effect is probably caused by soluble mediators, which partly pass into the systemic circulation.

CEA mRNA identification in peripheral blood is feasible for colorectal, but not for gastric or pancreatic cancer staging.

OBJECTIVE: It has been suggested that the molecular identification of cancer cells in the circulation may be useful in predicting the presence of micrometastasis in several cancer types. The aim of the present study was therefore to assess the feasibility of CEA mRNA identification in blood for diagnosing and staging colorectal, gastric and pancreatic cancer. METHODS: We studied 16 control subjects, 69 patients with colorectal (CRC), 30 with gastric (GC), 27 with pancreatic cancer (PC) and 8 with benign diseases of the pancreatobiliary tree. At diagnosis CEA mRNA was identified in peripheral blood by means of a RT-PCR procedure. RESULTS: The specificity of this test in control subjects was 94%, and its sensitivity in identifying CRC, GC and PC were 34, 37 and 41%, respectively. False-positive findings were recorded in 25% patients with benign diseases. No association was found between CEA mRNA and stage in patients with GC or PC. In CRC patients, positive CEA mRNA findings were correlated with local spread (chi(2) = 14.6, p<0.01), lymph node (chi(2) = 18.95, p<0.001) and distant metastasis (chi(2) = 11.3, p<0.001). In these cases, CEA mRNA, but not CEA, was entered in stepwise discriminant analysis to classify the presence of lymph node metastasis. CONCLUSIONS: The molecular detection of micrometastasis in the blood by means of CEA mRNA identification is feasible for colorectal, but not for gastric or pancreatic cancer staging. Further studies are needed in order to define the clinical utility of this marker also in follow-up protocols.

Proteases in gastrointestinal neoplastic diseases.

Cysteine and serine proteases are involved in cancer invasion and metastasis. In the past few years we investigated the tissue levels of these proteases in gastric cancer (GC), gastric precancerous changes (CAG), colorectal cancer (CRC) and the plasma and serum levels of proteases in several gastrointestinal tumours, using ELISA methods. Significantly higher antigen levels were found not only in GC tissue but also in CAG with respect to the levels found normal tissue; with respect to CAG, patients with dysplasia had higher levels than patients without dysplasia. The same findings were obtained in CRC. In general protease levels correlated with the major histomorphological parameters, such as grading and histotype in GC as well as in CRC. Tissue protease levels had a strong prognostic impact in GC, in which UPA was singled out by multivariate analysis as the major prognostic factor, and CRC. The plasma levels of urokinase-type plasminogen activator (UPA) and the serum levels of cathepsin B were significantly increased in patients with gastrointestinal tumours. In conclusions, cysteine and serine proteases may have a part not only in GC and CRC invasion and metastasis, but also in the progression of gastric precancerous changes into cancer. They are strong prognostic factors in GC and CRC. These proteases may also have a role as tumour markers in the early diagnosis of gastrointestinal tract tumours.

The role of cysteine and serine proteases in colorectal carcinoma.

BACKGROUND: Cathepsin B (CATB) and cathepsin L (CATL), which are cysteine proteases, urokinase-(UPA) and tissue-type plasminogen activator (TPA), both serine proteases, and their inhibitor type-1 (PAI-1) are believed to play an important role in colorectal carcinoma (CRC) invasion and metastasis. The objective of this study was to measure CATB, CATL, UPA, TPA, and PAI-1 in the same cancerous tissue (CANCER) and in tissues obtained from a tumor free area (NORMAL) to compare their respective prognostic roles in patients with CRC. METHODS: CANCER and NORMAL samples were obtained from 60 CRC patients undergoing surgery (36 males and 24 females; mean age, 63.8 years [range, 27-85 years]). The antigen concentrations were measured using an enzyme-linked immunoadsorbent assay method. The CANCER tissue also was examined in terms of major histomorphologic parameters such as differentiation, vascular invasion, degree of necrosis, and mucus production. RESULTS: Significantly higher antigen levels were found: 1) in CANCER versus NORMAL (with respect to CATL, UPA, and PAI-1, with significantly lower levels for TPA); 2) in CRC with versus without metastasis (CATB, CATL, and PAI-1); 3) in poorly versus well differentiated CRC (UPA and PAI-1); and 4) in advanced Dukes stages (PAI-1). CATB and CATL significantly correlated with UPA and PAI-1. Finally, CATL (P = 0.0001), UPA (P = 0.006), PAI-1 (P = 0.006), Dukes stage (P = 0.0001), presence of metastases (P = 0.003), and vascular invasion (P = 0.03) had a significant prognostic impact. CONCLUSIONS: The simultaneous up-regulation of cysteine and serine proteases in CRC confirms their role in colorectal tumor biology and particularly in the process of invasion and metastasis. Together with Dukes stage, determinations of CATL, UPA, and PAI-1 have a major prognostic impact in patients with CRC.

Analysis of Helicobacter pylori vacA and cagA genotypes and serum antibody profile in benign and malignant gastroduodenal diseases.

BACKGROUND: Helicobacter pylori species comprise different strains, cytotoxic and non-cytotoxic, which can be identified on the basis of their genomic pattern. AIMS: (1) To evaluate the polymorphism of the vacA gene and to ascertain whether the cagA gene is present in patients with gastric adenocarcinoma. (2) To study the anti-H pylori antibody profile using western blotting. PATIENTS: Twenty one patients with gastric adenocarcinoma and 71 with H pylori associated benign disease (nine gastric ulcer, 29 duodenal ulcer, 25 antral gastritis, and eight duodenitis). METHODS: The polymerase chain reaction was used to verify the presence or absence of cagA and to study the polymorphism of vacA in gastric mucosal samples obtained during endoscopy for patients with benign diseases and at surgery for patients with gastric adenocarcinoma. Fasting sera were used to assess anti-H pylori antibodies against different H pylori antigens by western blotting. RESULTS: CagA gene and the allele s1 of vacA were significantly less frequent in patients with antral gastritis (60% and 60%) compared with patients with gastric adenocarcinoma (94% and 100%) and with other non-malignant gastroduodenal diseases (93% and 87%) (chi 2 = 16.01, p < 0.001; and chi 2 = 13.97, p < 0.01). In patients with gastric adenocarcinoma, antibodies against a 74 kDa H pylori antigen were less frequently found than in patients with benign diseases. CONCLUSIONS: H pylori infection caused by cagA positive/vacA s1 strains is a frequent finding in patients with gastric adenocarcinoma. Prospective studies are needed to confirm whether the low incidence of positive serological response to the 74 kDa H pylori antigen in patients with gastric adenocarcinoma is important.

Urokinase-type plasminogen activator receptor in gastric cancer: tissue expression and prognostic role.

The urokinase-type plasminogen activator (UPA) and its inhibitor PAI-1 are thought to play an important part in gastric cancer (GC) invasion and metastasis. Little is known about the behavior and prognostic impact of the receptor for UPA (UPAR). The aims of the present study were: (1) to measure UPAR, UPA and PAI-1 levels in GC and in non-malignant tissue distant from the tumor (NORM); (2) to evaluate their relationship with histomorphological parameters; and (3) to determine their prognostic value. UPAR, UPA and PAI-1 levels were determined by ELISA in GC and NORM samples from 20 patients with GC undergoing surgery. The GC was also examined in terms of the presence (n = 10) or absence (n = 10) of metastasis, differentiation (five differentiated, 15 undifferentiated) and histotype. Survival was analysed using life table analysis. UPAR, UPA and PAI-1 were significantly higher in GC vs NORM, in the presence of metastasis (UPAR, UPA) and in undifferentiated GC (UPAR, PAI-1). UPAR significantly correlated with UPA and PAI-1. Low levels of UPAR (P = 0.04), UPA (P = 0.007) and PAI-1 (P = 0.02) were associated with a better survival. Our results demonstrate a sharp increase in UPAR in GC and suggest a prognostic role for it. The concomitant activation of UPAR, UPA and PAI-1 in GC confirm the important role of the plasminogen activator system in the process of invasion and metastasis.

N-terminal peptide of type III procollagen: a possible predictor of colorectal carcinoma recurrence.

BACKGROUND: The first step of colorectal carcinoma spread depends on the ability of the tumor cells to degrade and invade the extracellular matrix (ECM). The objectives of the current study were to evaluate the serum pattern of laminin, C-terminal peptide of Type I (PIP), and N-terminal peptide of Type III (PIIIP) procollagens, markers of ECM synthesis, in the follow-up of patients after resection for colorectal carcinoma and to evaluate their role in predicting local recurrence or metastases. METHODS: A total of 32 patients who had undergone resection for colorectal carcinoma were followed for a median period of 24 months (range, 6-36 months). Every 3 months, laminin, PIP, and PIIIP were measured in the sera together with the tumor markers carcinoembryonic antigen (CEA), CA 19-9, and tissue plasminogen activator (TPA). Twenty-one patients (Group 1) had no signs of recurrence, whereas the remaining 11 (Group 2) developed hepatic (n = 7) or pulmonary (n = 4) metastases. RESULTS: No variations were observed in either group for laminin, CEA, CA 19-9, or TPA, whereas significant increases in PIP and PIIIP were observed in both groups 3 months after surgery. The increase in PIP and PIIIP at the 3-month follow-up was significantly greater in Group 1 than in Group 2. The difference between values at 3 months and basal values enabled a discrimination between Group 1 and Group 2, with a sensitivity of 36% and 91% and a specificity of 71% and 71% for PIP and PIIIP, respectively. CONCLUSIONS: The authors believe PIIIP is useful as an early prognostic indicator of recurrence in the follow-up of patients who have undergone radical resection for colorectal carcinoma.

Neural cell adhesion molecule (N-CAM) in gastrointestinal neoplasias.

BACKGROUND: The neural adhesion molecule N-CAM, a membrane bound glycoprotein, seems to play an important role in the development of normal tissue architecture and in contact-dependent inhibition of cell growth. MATERIALS AND METHODS: We evaluated the behaviour of circulating N-CAM in patients with pancreatic cancer (24 cases) and chronic pancreatitis (15 cases) and compared it with that of 20 controls, 6 patients with colon adenoma, 31 with colorectal cancer or 21 with gastric cancer and ascertained the influence of tumor stage and grade on the findings. RESULTS: N-CAM levels were significantly lower in patients with pancreatic cancer and chronic pancreatitis than in the other groups studied. High levels were found only in few colorectal carcinoma patients (4/31). No correlation was found between tumor stage and N-CAM levels when pancreatic and colorectal cancer were considered. However, low N-CAM levels were found in 50% of advanced, but not in early gastric cancer. When pancreatic, colorectal and gastric cancer were considered, poorly differentiated (G3) had lower levels of N-CAM than well (G1) or moderately (G2) differentiated tumors. The variations found in circulating N-CAM were comparable to those in CEA but not to those in CA 19-9. CONCLUSIONS: a) perhaps because of its higher aggressiveness, pancreatic cancer is associated with low serum N-CAM levels unlike other gastrointestinal neoplasias; b) the association between aggressiveness and reduced N-CAM levels is borne out by the correlation found with the grade of differentiation; c) the reduced levels found in chronic pancreatitis suggest that this molecule plays a role in stromaparenchymal interactions, which might be altered in the presence of fibrotic phenomena.

Increased levels of cathepsin B and L, urokinase-type plasminogen activator and its inhibitor type-1 as an early event in gastric carcinogenesis.

BACKGROUND: Cysteine proteases [cathepsin B (CATB), cathepsin L (CATL)], the serine protease urokinase-type plasminogen activator (UPA) and its inhibitor type-1 (PAI-1) play an important part in cancer invasion. No data are available on the relationship between these proteases and gastric precancerous changes. AIMS: To determine CATB, CATL, UPA, PAI-1 in chronic atrophic gastritis, intestinal metaplasia and gastric epithelial dysplasia, as precancerous changes, and to compare these data with those obtained in gastric cancer. PATIENTS: Endoscopic biopsies were obtained from 12 patients with gastric cancer (cancerous tissue), 33 patients with chronic atrophic gastritis (all with intestinal metaplasia and 12 with dysplasia) and from 47 control subjects, for a total of 92 patients. METHODS: Antigen concentrations were measured using ELISA methods. Immunohistochemistry was performed using monoclonal anti-CATB and anti-PAI-1 antibodies. RESULTS: CATB, CATL, UPA and PAI-1 were significantly higher in chronic atrophic gastritis than in controls (CATB: P < 0.001; CATL: P < 0.005; UPA: P < 0.000001; PAI-1: P < 0.005). The same was observed for cancer. CATB and UPA were significantly higher in chronic atrophic gastritis, with versus without dysplasia (P < 0.05). Dysplastic epithelia showed strong immunoreactivity to PAI-1 and CATB. CONCLUSIONS: Our study demonstrates that cathepsins, UPA and PAI-1 may have a role not only in the process of cancer invasion, but also in the progression of precancerous changes into cancer.

Serum tumor markers in colorectal cancer staging, grading, and follow-up.

Early diagnosis of colorectal cancer, a frequent neoplasia in industrialized countries, permits curative surgery. In this study we assessed the clinical role of serum tumor markers determination in diagnosing, staging, and grading colorectal cancer; the role of carcinoembryonic antigen (CEA), CA 19-9, tissue polypeptide antigen (TPA) and CA 72-4 in colorectal cancer follow-up was also assessed. In 114 patients with colorectal cancer, the oncofetal antigen CEA was compared with the membrane-associated glycoproteins CA 19-9, CA 242, and CA 72-4 and with the cytokeratins TPA, tissue polypeptide-specific antigen (TPS) and tissue polypeptide monoclonal antigen (TPM). Overall, the most sensitive indices were TPA and TPS (67% and 70%, respectively). Tumor stage influenced the levels of CEA, CA 19-9, and TPA, but not those of TPS, while tumor grade influenced CEA and TPS, but not CA 72-4, TPA, and TPM. TPA was the most sensitive index in identifying early or well-differentiated colorectal cancers. The sensitivity was enhanced when this marker was determined in combination with CEA, in diagnosing both advanced and early colorectal tumors. Seventy-seven patients were followed up after therapy for at least 18 months. CEA was the most sensitive index of recurrence (58%); however, this sensitivity is too low to consider tumor markers useful in colorectal cancer follow-up.

Serum tumor markers in monitoring patients: interpretation of results using analytical and biological variation.

During cancer monitoring, data on biological and analytical variation are required in order to define the critical difference which provides an objective means to interpret serial values. We evaluated four tumor markers on serial samples collected from healthy subjects and patients. Analytical coefficients of variation (CV(A)), were obtained from "precision profiles" based on the differences between duplicates cumulated from assay runs in the laboratory. We defined the mean intrasubject biological variation (CV(I)) for CA 19-9 and TPA, separately for healthy people and patients; since the differences between the two groups were not statistically significant, we pooled the results and re-evaluated CV(I) in the combined groups (CA 19-9: CV(I) = 15.9%; TPA: CV(I) = 25.7%). In addition, we evaluated CV(I) for CEA (10.9%) and for TPS (25.9%) in patients. We then evaluated the inter-subject biological variations (CV(G)); the calculated indices of individuality for the four markers were less than 0.6 which shows conventional reference values to be of little utility for interpretation. We finally evaluated the critical differences (p < 0.05) for CA 19-9 (CD = 44.7%), for TPA (CD = 72.5%), CEA (CD = 32.7%) and TPS (CD = 72.5%); these are generally applicable since there was no heterogeneity in intra-subject biological variability.

[Prognostic role of cisteine and serin proteases in gastriC cancer]. / Cisztein-és szerin-proteázok prognosztikai jelentösége gyomorrákban.

Cysteine proteases (cathepsin B and L), the serine protease urokinase-type plasminogen activator and its inhibitor type-1 play an important part in cancer invasion and metastasis. The authors determined the protease concentrations in gastric cancer tissues, using the ELISA method, in patients with gastric cancer. They evaluated the prognostic role of proteases and the relationship that these proteases may have with other histomorphological prognostic parameters such as tumor staging, grading, histotype, Borrmann classification. The Cox survival analysis showed that cathepsin B (p = 0.002), urokinase-type plasminogen activator (p = 0.0001) and the inhibitor type-1 (p = 0.0004) significantly correlated with poor prognosis. The tumor staging, grading, Borrmann classification correlated also significantly with survival time. Urokinase-type plasminogen activator was selected as the single independent variable in the Cox model (p = 0.0001).

Cysteine and serine proteases in gastric cancer.

BACKGROUND: Cysteine proteases (cathepsin B [CATB] and cathepsin L [CATL]), the serine protease urokinase-type plasminogen activator (UPA), and plasminogen activator inhibitor type-1 (PAI-1) are thought to play an important part in cancer invasion and metastasis. The aims of this study were to measure CATB, CATL, UPA, and PAI-1 in gastric cancer (GC) and normal mucosa distant from the tumor (NORM); to evaluate whether tissue levels are related to tumor stage, grade, or histotype; to assess their prognostic relevance; and to examine UPA and PAI-1 expression immunohistochemically. METHODS: Gastric cancer and NORM samples were obtained from 25 patients with gastric cancer patients undergoing surgery (17 males, 8 females; mean age, 62 years; range, 31-84 years). Antigen concentrations were measured using the enzyme-linked immunosorbent assay method. Immunohistochemistry was performed using monoclonal UPA and PAI-1 antibodies. RESULTS: Significantly higher antigen levels were found: (1) in GC vs. NORM (CATB, CATL, UPA, PAI-1) tissues; (2) in GC with versus without metastasis (CATB, CATL, UPA); (3) in poorly or moderately versus well differentiated GC; and (4) in diffuse versus intestinal-type GC (CATB, CATL). Urokinase-type plasminogen activator, PAI-1 and CATB levels had a significant prognostic impact. Cancer and stromal cells, showed immunoreactivity to anti-UPA and anti-PAI-1 antibodies. CONCLUSIONS: These results confirm the important role of CATB, CATL, UPA and PAI-1 in gastric cancer progression. Higher levels are detected in GC with metastases, poorer differentiation, and diffuse histotype, thus identifying patients with a worse prognosis.

[Role and behavior of cathepsin B and cathepsin L in gastric cancer]. / A katepszin B és a katepszin L viselkedése és szerepe a gyomorrákban.

Cathepsin B and cathepsin L--cysteine proteinases--may play an important role in cancer invasion and metastasis. The authors determined tissue antigen concentrations of cathepsins, using the ELISA method, in 25 patients with gastric cancer (17 males, 8 females, mean age 62, range 31-84). They evaluated the possible relationship that these proteinases may have with the presence of metastases, differentiation and histotype. Significantly higher cathepsin B and cathepsin L antigen levels were found: 1. in gastric cancer tissues vs. normal tissues distant from tumors (CATB: p < 0.05, CATL: p < 0.005); 2. in diffuse vs. intestinal type cancers (p < 0.05); 3. in patients with poorly vs. well-differentiated cancers (p < 0.05); in gastric cancers with vs. without metastasis (p < 0.05). Their results confirm that cathepsin B and L play an important role in gastric cancer invasion and metastasis. Considering the significantly higher cathepsins detected in cancers with metastasis, a poor differentiation and of diffuse histotype, these proteinases could be useful for identification gastric cancer patients with a poor prognosis.

The role of urokinase-type plasminogen activator and its inhibitor PAI-1 in gastric cancer.

BACKGROUND: It has been proposed that the serine protease urokinase-type plasminogen activator (UPA) and its inhibitor type-1 (PAI-1) may play an important part in cancer spread and metastasis. AIMS OF THE STUDY: 1. To determine the UPA, PAI-1 antigen levels in gastric cancer (GC) and in normal gastric mucosa far from the tumor (NORM). 2. To evaluate changes related to the presence of metastasis, tumor differentiation (grading) and histotype. PATIENTS AND METHODS: Samples of tumor and of normal mucosa were obtained from 25 GC patients undergoing surgery (17 males, 8 females, mean age 62, range 31-84). Antigen concentrations were measured using the ELISA method. RESULTS: Significantly higher UPA and PAI-1 antigen levels were found: 1. in GC vs. NORM tissue; 2. in patients with poorly and moderately differentiated GC vs. well-differentiated GC. UPA was significantly higher in GC with vs. without metastasis. PAI-1 was also higher, though not significantly so, in GC with metastasis than in GC without metastasis. CONCLUSION: Our results confirm that the serine protease UPA and its inhibitor PAI-1 play an important role in GC progression. Considering the higher protease levels detected in GC with metastases and poorer differentiation, UPA and PAI-1 might be useful for identifying gastric cancer patients with a poor prognosis.