Clinical significance of toll-like receptor 3, 4, and 9 in gastric cancer.

Toll-like receptors (TLRs) have raised an extraordinary interest in cancer research due to their role in tumor progression. By activating the production of several biological factors, TLRs drive an inflammatory response and activate the adaptive immune system. The aim of this study was to investigate the expression and clinical relevance of TLR3, TLR4, and TLR9 in gastric cancer. For this purpose, an immunohistochemical study on cancer specimens from 106 patients with gastric cancer was performed using tissue arrays and specific antibodies against TLR3, TLR4, and TLR9. The results indicate that gastric carcinomas samples show high expression of TLR3, TLR4, and TLR9 by cancer cells. The expression of TLR3 by cancer cells was significantly associated with a poor overall survival in patients with resectable tumors. Moreover, in patients with resectable tumors and lymph node invasion, a high TLR3 expression defines a population with even worse prognosis. Therefore, TLR3 may have clinical interest as indicator of tumor aggressiveness and as a prognostic indicator in gastric cancer.

Toll-like receptor-4 expression by stromal fibroblasts is associated with poor prognosis in colorectal cancer.

Because of the important role in inflammation and tissue regeneration, toll-like receptors (TLR) are likely candidates to mediate effects of the innate immune system on tumorigenesis. The aim of this study was to investigate the expression and clinical relevance of TLR in colorectal cancer (CRC). The expressions of TLR3, TLR4, TLR7, and TLR9 were analyzed in 104 patients with resectable CRC by immunohistochemistry. The evaluation of the expression consisted on measuring the overall level of TLR expression and by each cell type. The results showed a direct association between the histologic grade of tumor and TLR9 expression by tumor cells. TLR4 expression by tumor cells was significantly associated with a lower rate of tumor recurrence, whereas the expression by fibroblasts was significant and independently associated with a high rate of tumor recurrence and with a shortened overall survival in patients; particularly in tumors from left colon and rectum. Therefore, TLR4 expression by fibroblasts could be a useful prognostic marker in CRC.

Effect of the expression of matrix metalloproteases and their tissue inhibitors on survival of patients with resectable colorectal cancer.

BACKGROUND: Matrix metalloproteases (MMPs) and their tissue inhibitors (TIMPs) are of crucial importance in the degradation of the stromal connective tissue and basement membrane components. Study of the behavior of these components might help to predict the aggressiveness of tumors. AIMS: To evaluate the expression and clinical relevance of MMPs and TIMPs for patients with resectable colorectal carcinoma. METHODS: An immunohistochemical study was performed using tissue arrays and specific antibodies against MMPs-1, 2, 7, 9, 11, 13, and 14, and TIMPs-1, 2 and 3. Determinations were performed in cancer specimens from 104 patients with resectable colorectal cancer. The minimum period of follow-up was 12.5 years for patients without recurrence. To identify specific groups of tumors with distinct expression profiles, the data were analyzed by unsupervised hierarchical cluster analysis. RESULTS: Expression of MMP-11 by fibroblasts and MMP-13 by tumor cells were associated with poor prognosis. The dendrogram revealed first-order division of tumors into two distinct MMP/TIMP molecular profiles, designated group 1 (n = 50) and group 2 (n = 54). Group 2 was characterized by significantly higher expression of MMP-1, 11, and 13, and TIMP-3. CONCLUSION: Our results emphasize the prognostic value of MMP-11 and 13 expression in colorectal cancer.

Lymphatic and/or blood vessel invasion in gastric cancer: relationship with clinicopathological parameters, biological factors and prognostic significance.

BACKGROUND: Lymphatic and/or blood vessel tumoral invasion (LBVI) is a common histopathologic finding of gastric carcinomas, which could make it an additional cost efficient marker and help in the detection of patients at risk for recurrence. MATERIALS AND METHODS: The subjects of this study were 144 patients with primary gastric adenocarcinoma, who consecutively underwent surgery. LBVI was evaluated by H&E staining and complementary with immunohistochemical staining with anti-CD34. Intratumoral levels of EGFR were analyzed with a radioligand technique, whereas c-erbB-2 and tPA were determined by ELISA methods; pS2, cathepsin D and hyaluronic acid by immunoradiometric assays; and VEGFR-1 and -2 by immunohistochemical assays. The mean follow-up period for these patients was 33.1 months. RESULTS: LBVI was present in 46 patients (31.9%). The presence of LBVI correlated significantly with tumor stage, lymph node involvement, surgical resectability, histological type and histological grade, being present in a higher percentage among II-IV tumor stage (P = 0.0001), poorly differentiated (P = 0.01), diffuse type (P = 0.009), R1-R2 (P = 0.002) and lymph node-positive (P = 0.005) tumors. In addition, statistical analysis demonstrated that LBVI was significantly associated with a poorer overall patients' survival in the univariate analysis (P = 0.0001) as well as in the multivariate analysis (P = 0.009). However, our results failed to show any significant relationship between LBVI and any of the intratumoral biological parameters studied. CONCLUSION: LBVI provides additional useful information that could be applied to identify gastric cancer patients at risk for recurrence, who might be candidates for further adjuvant therapies.

Expression and clinical signification of cytosolic hyaluronan levels in invasive breast cancer.

BACKGROUND: Hyaluronic acid (HA), a high-molecular weight glycosaminoglycan, has been considered to be involved in the growth and progression of malignant tumors in several experimental studies. The objective of this work was to evaluate the cytosolic HA content in breast cancer, its possible relationship with clinicopathological tumor parameters and steroid receptor status, as well as its potential prognostic significance. METHODS: Cytosolic HA levels were examined by means of immunoradiometric techniques in 850 patients with invasive breast cancer. The mean follow-up period for these patients was 55.1 months. RESULTS: Cytosolic HA levels ranged widely in tumors (4-59767 ng/mg protein; median: 4960). Statistical analysis showed that HA levels were significantly higher in younger patients (p=0.0001), as well as in premenopausal than in postmenopausal patients (p=0.001). HA levels were also significantly higher in ductal or lobular histological type than in other histological types (coloid, medullar or papillar types) (p=0.0001). Likewise, HA correlated significantly and positively with tumoral levels of PgR (r sub S: 0.11; p=0.001) in the all group of patients. In the subgroup of patients with ductal invasive type, HA levels were also significantly higher in well differentiated tumors and in diploid tumors. In addition, in this latter group of patients, HA levels in tumors correlated also positively and significantly with the either estrogen-inducible proteins: PgR (r sub S: 0.11; p=0.001), pS2 (r sub S: 0.117; p=0.008) and tPA (r sub S: 0.314; p=0.0001). On the other hand, significant association between HA intratumoral levels and relapse-free survival and overall survival in the overall group of patients was not found. However, high HA intratumoral levels were significantly associated with longer relapse-free survival in the subgroup of patients with ductal histological type tumors (p=0.01), as well as in those patients without any type of systemic adjuvant treatment (p=0.01). CONCLUSIONS: Our results suggest that high intratumoral levels of HA may be associated with tumors of favorable evolution in certain subgroups of patients with breast cancer. Thus, HA may provide additional prognostic information to that given by other biochemical markers currently used in breast cancer.

Clinical significance of the quantitative assessment of the cytosolic concentration of HER-2/neu protein in breast cancer by immunoenzymatic assay (ELISA).

PURPOSE: Retrospective analysis to assess the prognostic and predictive value of HER-2/ neu expression in breast tumors, quantified by enzyme immunoassay (ELISA). METHODS: Quantification of HER-2/neu was performed on cytosolic extracts from 914 cases of primary invasive breast carcinomas. Relapse-free and overall survival data were available from 889 patients. The prognostic value of HER-2/neu levels was assessed considering them as a continuous, dichotomic or quartile variable. RESULTS: Cytosolic HER-2/neu levels ranged widely in breast carcinomas (median: 746.5 NHU/mg; range: 2.8-80,000 NHU/mg protein). HER-2/neu protein levels were significantly higher in either moderately or poorly differentiated tumors, as well as in those showing a ductal histological type, aneuploidy or a high S-phase fraction. There was a significant and positive association between cytosolic and membranous HER-2/neu levels (n=162, r sub S=0.53; P<0.0001). In addition, cytosolic HER-2/neu level correlated weakly with progesterone receptors but not with estrogen receptors. Elevated cytosolic HER-2/neu levels (> or =1,400 NHU/mg protein) were associated with a high probability of both shortened relapse-free survival and overall survival. This same cut-off value was obtained when we divided the overall group of patients in a training set. However, this HER-2/neu value did not achieve any statistical significance in a validation set used to make sure that the cut-off was correct. Nevertheless, when we divided the obtained data into three different groups with respect to the quartile values (Q) of the intratumoral oncoprotein levels (< or = Q1 vs Q1-Q2 vs > Q3), we observed that patients with either low HER-2/ neu levels (< or = Q1) or high HER-2/neu levels (> Q3) had shorter both relapse-free survival and overall survival curves than those patients with intermediate HER-2/neu levels. On the other hand, high HER-2/neu levels predicted a poor response to adjuvant chemotherapy but not to adjuvant hormonal therapy with tamoxifen. CONCLUSIONS: The results of the present investigation indicate that by quantitatively determining the content of HER-2/neu oncoprotein, groups of high-risk breast cancer patients could be identified, for a more effective clinical management.

Tissue-type plasminogen activator (tPA) in breast cancer: relationship with clinicopathological parameters and prognostic significance.

BACKGROUND: Tissue-type plasminogen activator (tPA) is a serine protease primarily involved in the intravascular dissolution of blood clots. High intratumoral tPA levels are associated with prognosis in several human tumors. In addition, tPA has been shown to be an estrogen-inducible protein in human breast cancer cell lines. The aim of the present study was to analyze the cytosolic tPA content in primary breast carcinomas and its potential clinical value. MATERIALS AND METHODS: tPA was measured by a solid-phase enzyme immunoassay in tumor cytosol samples obtained from 800 patients with breast cancer. The median follow-up period was of 49.2 months. RESULTS: Cytosolic tPA levels ranged widely in breast carcinomas (median: 3.9; range: 0.1- 315.3 ng/mg protein). tPA levels were significantly lower in large tumors, as well as in those showing poor differentiation, estrogen (ER) or PgR-negativity, aneuploidy, or a high S-phase fraction. In addition, low tPA intratumoral levels were associated with a high probability of both shortened relapse-free and overall survival in all patients and in the subgroup with node-negative tumors. However, our results did not show any significant relationship between intratumoral tPA levels and prognosis in the different subgroups of patients, stratified according to the type of systemic adjuvant therapy received (chemotherapy, tamoxifen or chemotherapy plus sequential tamoxifen). CONCLUSION: The results of the present investigation indicate that low intratumoral tPA levels are associated with aggressiveness and poor prognosis in breast cancer patients. However, the study suggests that tPA levels do not predict response to systemic adjuvant therapy.

Expression and clinical significance of CD44V5 and CD44V6 in resectable colorectal cancer.

PURPOSE: This study was conducted to evaluate the prognostic significance of CD44v5 and CD44v6 in resectable colorectal cancer. MATERIALS AND METHODS: Membranous CD44v5 and CD44v6 levels were measured by an immunoenzymatic assay in tumors and surrounding mucosal samples obtained from 105 patients with resectable colorectal carcinomas. RESULTS: There were no significant differences of CD44v5 levels between tumors [median: 3.2 (range: 0.9-83.5) ng/mg protein) and surrounding mucosal samples (3 (3-146.2) ng/mg protein]. However, tumor samples showed significantly higher CD44v6 levels [19.5 (2.2-562.9) ng/mg protein] than mucosal samples [5 (5-230) ng/mg protein] (P=0.0001). Patients with higher CD44v5 or CD44v6 content in tumor samples had a considerably shorter relapse-free survival (P<0.05, for both). Patients with a higher CD44v6 content also had a shorter relapse-free and overall survival in the multivariate analysis (P<0.05). CONCLUSION: The results of this study suggest a role of CD44v5 and CD44v6 in colorectal cancer progression. Membranous CD44v levels in primary tumors, measured by immunoenzymatic assay, may contribute to a more precise prognostic estimation in patients with resectable colorectal cancer.

CD44s expression in resectable colorectal carcinomas and surrounding mucosa.

BACKGROUND: CD44s (standard isoform) is a cell adhesion molecule belonging to the family of the hyaluronan-binding proteins. The CD44 family has been found to be overexpressed in epithelial tumors, where they are generally in relationship with tumor growth and metastasic properties. The aim of this work was to evaluate the membranous CD44s content in colorectal cancer and in healthy surrounding mucosa, its possible relationship with clinicopathological parameters, and its potential prognostic significance. MATERIALS AND METHODS: Membranous CD44s levels were measured by an immunoenzymatic assay in tumors and surrounding mucosa samples from 72 patients with resectable colorectal carcinomas. The patients were followed for a mean time period of 30 months. RESULTS: There was a wide variability of CD44s levels in tumor-surrounding mucosal samples (26.6-727 ng/mg protein) as well as in tumors (28.5-381 ng/mg protein). Tumor samples showed significantly higher CD44s levels (median: 99.1 ng/mg protein) than surrounding mucosal samples (81 ng/mg protein) (p=0.03). In the same way, CD44s levels in tumors as well as in surrounding mucosal samples were significantly higher in high S-phase tumors than in low S-phase tumors (p=0.001 for both). There was no significant relationship between tumor CD44s levels and patient's outcome. However, high levels of the glycoprotein in nonneoplastic surrounding mucosa were significantly (p=0.018) associated with a poor overall patient survival. CONCLUSION: CD44s may play a role in the tumorogenesis of colorectal carcinomas. In addition, CD44s levels in tumor-surrounding mucosa may provide, in concert with some clinicopathological parameters, important information about prognostic evaluation of patients with resectable colorectal carcinomas.

Expression and prognostic significance of lysozyme in male breast cancer.

BACKGROUND: Lysozyme, one of the major protein components of human milk that is also synthesized by a significant percentage of breast carcinomas, is associated with lesions that have a favorable outcome in female breast cancer. Here we evaluate the expression and prognostic value of lysozyme in male breast cancer (MBC). METHODS: Lysozyme expression was examined by immunohistochemical methods in a series of 60 MBC tissue sections and in 15 patients with gynecomastia. Staining was quantified using the HSCORE (histological score) system, which considers both the intensity and the percentage of cells staining at each intensity. Prognostic value of lysozyme was retrospectively evaluated by multivariate analysis taking into account conventional prognostic factors. RESULTS: Lysozyme immunostaining was negative in all cases of gynecomastia. A total of 27 of 60 MBC sections (45%) stained positively for this protein, but there were clear differences among them with regard to the intensity and percentage of stained cells. Statistical analysis showed that lysozyme HSCORE values in relation to age, tumor size, nodal status, histological grade, estrogen receptor status, metastasis and histological type did not increase the statistical significance. Univariate analysis confirmed that both nodal involvement and lysozyme values were significant predictors of short-term relapse-free survival. Multivariate analysis, according to Cox's regression model, also showed that nodal status and lysozyme levels were significant independent indicators of short-term relapse-free survival. CONCLUSION: Tumor expression of lysozyme is associated with lesions that have an unfavorable outcome in male breast cancer. This milk protein may be a new prognostic factor in patients with breast cancer.