Polyadenylate polymerase enzymatic activity in mammary tumor cytosols: A new independent prognostic marker in primary breast cancer.

Polyadenylate polymerase (PAP) is one of the enzymes involved in the formation of the polyadenylate tail of the 3' end of mRNA. High levels of PAP activity were associated with rapidly proliferating cells. Here we evaluate the prognostic value of PAP activity in breast cancer patients. PAP specific activity values were measured by a highly sensitive assay in the tumor cytosols of 228 women with primary breast cancer. The median follow-up period was 58 months. PAP specific activity values ranged from 2.1-39.4 units/mg protein in the breast tumor cytosols, and the activity was correlated with the level of expression of the antigen. An optimal cutoff value of 5.5 units/mg extracted protein was first defined by statistical analysis. PAP status was then compared with other established prognostic factors in terms of relapse-free survival (RFS) and overall survival (OS). PAP activity levels had a tendency to increase with tumor-node-metastasis (TNM) stage and were higher in node-positive patients. Evaluation of the prognostic value of PAP was performed using univariate and multivariate analyses. Univariate analysis showed that PAP-positive patients had a less favorable prognosis for both RFS (relative risk (RR) = 2.35; P < 0.001] and OS (RR = 3.15; P < 0.001). PAP significantly added to the prognostic power for RFS (RR = 2.51; P = 0.0012) and OS (RR = 4.21; P < 0.001) in multivariate analysis, whereas patient age, tumor size, and nodal and ER status remained independent factors for predicting survival. When only node-negative patients were examined, PAP was found to be an independent factor for predicting RFS (RR = 3.68; P = 0.0032) and OS (RR = 4.81; P < 0.001). PAP did not appear to have a prognostic significance for node-positive patients. PAP is a new prognostic factor for early recurrence and death in breast cancer patients. Our results suggest that PAP may be used as an independent unfavorable prognostic factor in node-negative breast cancer patients because there were no significant associations between PAP and the other prognostic indicators evaluated in this group of patients.

Predictive value of c-erbB-2 and cathepsin-D for Greek breast cancer patients using univariate and multivariate analysis.

The value of various prognostic factors in breast cancer patients has been determined in a number of studies. One hundred thirty-eight Greek women were followed up over a 5-year period after surgery for breast cancer. Amplification and overexpression of c-erbB-2 was found in 22.4% and 29.7% of the respective cases, and the concentration of total cytosolic Cathepsin-D (CD) in 46.4% of them was high (> or = 60 pmol/mg protein). The examined biological variables were compared with standard clinicopathological prognostic factors for the disease and related to early relapse (ER; before 3 years), relapse-free survival (RFS; median, 5 years), and overall survival (OS; median, 5 years). It was found that high CD levels significantly shorten ER of both node-negative and node-positive patients (P < 0.0001 and P = 0.002, respectively) and have prognostic value for RFS and OS of node-negative patients (P = 0.0012 and P = 0.0288, respectively), but lose their value as relapse predictors for node-positive patients for periods longer than 3 years. Overexpression of c-erbB-2 was found to be predictive for OS of node-positive and -negative patients (P = 0.0048 and P = 0.0285, respectively), but its predictive power was weak for ER (P = 0.0456) and RFS (P = 0.0455) of node-negative patients and disappeared for node-positive patients. c-erbB-2 amplification offers minimal assistance to the prediction. In conclusion, high CD concentration is indicative of ER of patients, and c-erbB-2 overexpression correlates with OS of patients.

HER-2/neu overexpression in breast cancer: an immunohistochemical study including correlations with clinicopathologic parameters, p53 oncoprotein and cathepsin-D.

PURPOSE: To evaluate the relationship between HER-2/neu overexpression and standard as well as investigational prognostic factors in Greek females with invasive breast carcinoma. MATERIALS AND METHODS: Paraffin-embedded tumor sections from 128 consecutive primary breast cancer patients were screened for HER-2/neu oncoprotein (p185) overexpression by immunohistochemistry using the polyclonal antibody A 0485. The relationship between HER-2/neu staining and other established markers of prognosis were examined using both a univariate and a multivariate analysis. RESULTS: HER-2/neu overexpression was detected in 46.1% of tumors. No statistical correlation was found between HER-2/neu and age, tumor diameter, histologic type, nodal status, tumor grade, stage, estrogen and progesterone receptor status or cathepsin-D. A significant correlation was noted between HER-2/neu and p53 overexpression in the total sample (p=0.014) as well as in node-positive patients (p=0.026). In those groups, HER-2/neu and p53 co-expression was found in 19.5% and 21.6% of cases, respectively. In node-negative patients, HER-2/neu overexpression did not correlate with any other marker. CONCLUSION: HER-2/neu overexpression seemed to correlate only with p53 oncoprotein.

Cathepsin-D and c-erb-B 2 have an additive prognostic value for breast cancer patients.

In breast cancer, axillary lymph node invasiveness is the major prognostic factor in predicting relapse and metastasis. Nevertheless, since 30% of node-negative tumours also relapse, it is necessary to develop other independent prognostic factors. Oncogene amplification and overexpression as well as the level of cathepsin-D have been proposed as additional prognostic factors. Recent studies suggest that the acidic lysosomal proteinase cath-D, present in all cells and known to be secreted in breast cancer cells, may be implicated in the process of tumour invasion and metastasis. We have compared the cytosolic cath-D level with the amplification and the overexpression of the oncogenes c-myc and c-erb-b-2 in 62 breast carcinomas (52 primary and 10 metastatic). Using a cut-off level of 60 pmol/mg protein, the status of cath-D showed a positive correlation with c-myc amplification (P = 0.01) or overexpression (P = 0.02). In contrast, no correlation was found between cath-D and c-erb-B-2 amplification or overexpression. Also, no correlation was found between cath-D and established prognostic factors such as node invasiveness, steroid receptors, grade and menopausal status. Nevertheless, a weak correlation was found between cath-D levels and tumour status (P = 0.05). In conclusion, in breast cancer, a high cytosolic cath-D concentration is more frequent in tumours with c-myc amplification and overexpression but is dissociated from c-erb-B-2 amplification or overexpression, suggesting that the determination of cath-D, as well as the latter two markers, will have an additional prognostic value.

c-erbB-2 overexpression may be used as an independent prognostic factor for breast cancer patients.

Insight into correlations between specific gene amplification and the clinical behavior of tumors may provide new prognostic tools High c-erbB-2 expression is an early feature in some breast tumors, whereas c-myc may be involved in the development of neoplasia. After an initial flurry of excitement about their prognostic significance, controversy has arisen about their independent importance. In an attempt to solve this problem, we decided to study c-erbB-2 and c-myc amplification and overexpression in 62 unselected breast carcinomas. This was done in order to correlate them statistically with one another, as well as with other prognostic parameters. A positive correlation was discovered between c-erbB-2 amplification and overexpression (P = 0.02); however, the correlations between c-erbB-2 amplification and c-myc amplification and overexpression (P = 0.06 and P = 0.095 respectively) were found to be negative. In addition, no correlation was found to exist between c-erbB-2 amplification and Cathepsin-D, steroid receptors, node status and menopausal status, as well as between c-erbB-2 overexpression and Cathepsin-D, node invasiveness, tumor status, grade or menopausal status. In conclusion, the c-erbB-2 overexpression has positive correlation with only a few other prognostic parameters, and therefore can be used as an independent prognostic factor.

Poly(A)polymerase activity levels in breast tumour cytosols.

The enzyme poly(A) polymerase (PAP) catalyses the polyadenylation of mRNA and its activity levels vary within the cell cycle. The levels of activity of this enzyme were measured in the cytosol of breast tumours from 62 untreated patients and compared to clinical prognostic parameters as well as other biological markers. The enzyme levels measured ranged from 3 to 46 units/mg protein. A statistically significant association was observed between high PAP activity values and the TNM stage of the disease as well as node invasiveness. Furthermore, there was a positive correlation between PAP activity values and c-erbB-2 overexpression but not with its amplification. No significant correlation was observed with c-myc amplification or overexpression and cathepsin D levels. A direct relationship between steroid receptor content and PAP activity levels, which was more prominent in the case of the progesterone receptor, was observed. However, also on the basis of previous data PAP activity may prove to be indicative of aggressive disease. Furthermore, measurements of PAP activity may contribute to the definition of the biological profile of tumour cells.

Significance of urokinase-type plasminogen activator and plasminogen activator inhibitor-1 (PAI-1) expression in human colorectal carcinomas.

Despite the advances in the medical care of colorectal carcinoma patients, the prognosis has improved only marginally over recent decades. Thus, additional prognostic indicators would be of great clinical value to select patients for adjuvant therapy. In the present study, the antigen levels of urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1, and their immunohistochemical staining were compared in paired colorectal tumor (n = 64) and background colon tissue of the same patients with clinical and pathological staging. The antigen levels, measured with an ELISA method, were found to be significantly higher in cancer tissue (mean 1.92 ng/mg protein for uPA and 7.08 for PAI-1) than in corresponding normal mucosa (0.29 ng/mg protein for uPA and 1.11 ng/mg protein for PAI-1). There was a positive correlation between uPA and PAI-1 antigen levels and clinicopathological parameters such as grade (p < 0.001 and p = 0.01, respectively), while for Dukes' stage, only PAI-1 correlated positively (p = 0.018). Nodal status correlated positively with uPA but not with PAI-1 antigen levels. Immunohistochemical localization of both antigens was observed mainly in cancer cells and much less in stromal cells. Staining intensity increased from adenoma to adenocarcinoma. The degree of staining was associated with grade, Dukes' stage and nodal status for uPA (p < 0.001, p = 0.002, p < 0.001, respectively) and only with grade for PAI-1 (p = 0.007).

Determination of c-myc amplification and overexpression in breast cancer patients: evaluation of its prognostic value against c-erbB-2, cathepsin-D and clinicopathological characteristics using univariate and multivariate analysis.

C-myc and c-erbB-2 amplification and/or overexpression as well as total cathepsin-D (CD) concentration have been reported to be associated with poor prognosis in breast cancer. The prognostic significance, however, remains somewhat controversial, partly because of discrepancies among the different methodologies used. We determined the amplification and overexpression of c-myc oncogene in 152 breast cancer patients and examined its prognostic value in relation to c-erbB-2 amplification and overexpression, high concentration of CD (> or = 60 pmol mg(-1) protein) and standard clinicopathological prognostic factors of the disease. High CD concentration, as well as c-myc amplification and overexpression, proved to be the best of the new variables examined for prediction of early relapse (ER; before 3 years). After multivariate analysis only CD remained significant, which suggests that the prognostic power of these variables is similar. Using univariate analysis we proved that c-myc amplification and overexpression were highly significant for disease-free survival (DFS) (P = 0.0016 and P = 0.0001 respectively) and overall survival (OS) (P < 0.0001 and P = 0.0095 respectively), although by multivariate analysis c-myc overexpression was statistically significant only for DFS (P = 0.0001) and c-myc amplification only for OS (P = 0.0006). With regard to c-erbB-2, only its overexpression appeared to be significant for DFS and OS, although after multivariate analysis its prognostic power was weaker (P = 0.030 and P = 0.024 respectively). c-myc amplification and overexpression exhibited a tendency for locoregional recurrence (LRR) (P = 0.0024 and P = 0.0075 respectively), however, their prognostic value was lower after multivariate analysis and only CD remained significant.