The levels of cathepsins (Cats) B, H, and L and their inhibitors stefin A and cystatin C were determined in the sera of 43
patients with metastatic melanoma, in 54 patients with treated cutaneous ...melanoma with no evidence of metastatic disease,
and in 30 healthy blood donors, using quantitative ELISAs. The levels of Cats B and H and cystatin C were significantly higher
within the group of metastatic melanoma patients compared with the healthy controls. The median Cat B was 4.8 versus 3.6 ng/ml
(P < 0.013), the median Cat H was 13.7 versus 4.9 ng/ml (P < 0.0001), and the median cystatin C was 470 versus 320 ng/ml (P
< 0.02). Cat H was also significantly increased within the group of melanoma patients with no metastasis, with a median of
9.6 ng/ml. Cat B was found to correlate with Cat L (r = 0.36; P < 0.02) and cystatin C (r = 0.41; P < 0.008). The serum level
of Cat H was significantly increased in patients showing no response to the chemoimmunotherapy as compared to the level in
responders. Metastatic melanoma patients with high contents of Cat B and Cat H experienced significantly shorter overall survival
rates than the patients with low levels of each enzyme (Cat B: P < 0.003 and relative risk, 2.5; Cat H: P < 0.006 and relative
risk, 2.4, using medians as cutoff values). The other potential factors for prognosis for this group of patients revealed
moderate (histological type and age) or no (tumor thickness, sex, and lymph node metastasis) prognostic significance. Similarly,
no difference in survival was found for stefin A, cystatin C, and Cat L. These results suggest that the serum levels of Cats
B and H could serve as prognostic factors for patients with advanced melanoma.
None of the established prognostic factors in breast cancer (BC) is able to determine the final outcome with certainity. Tumor biological factors involved in tumor invasion and metastasis, such as ...cathepsins and proteins of u-PA system, have been put forward in the recent literature as strong novel prognostic factors in BC. We therefore evaluated prognostic and predictive value of cathepsin-D (CD) and cathepsin-L (CL) in 715 operable BC patients. CD and CL were determined in tumor extracts using immunoradiometric and ELISA assays, respectively. During follow-up (median 37 months), 151 (21%) patients relapsed. In a multivariate analysis of disease-free survival (DFS), CL (p=0.04), nodal status (p<0.001) and hormone receptor status (p<0.001) were the only independent significant prognostic factors. CL thus provided independent prognostic information on DFS and could also predict a response to adjuvant chemotherapy (ChT), while CD had no significant prognostic and predictive impact.
The present study on the prognostic and predictive value of serine proteases was conducted in 460 early breast cancer patients mostly treated with some kind of adjuvant systemic therapy: 156 received ...chemotherapy, 141 hormone therapy and 111 a combination of both. Already in univariate analysis PAI-1 was the only proteolytic factor with a significant impact on DFS, which was retained in multivariate analysis (p = 0.020); PAI-2 showed borderline significance in univariate analysis (p = 0.0503) and uPA did not present as a significant prognostic factor for DFS in our patient series. In a separate univariate analysis of DFS on patient subgroups defined by adjuvant systemic therapy, a higher risk of relapse associated with higher uPA and PAI-1 levels was found in the subgroup of patients who did not receive any treatment; this difference did not reach the level of significance, probably due to the small number (n = 52) of patients in this group (HR 1.37; p = 0.71 and HR 2.14; p = 0.321, respectively). A higher risk of relapse was also found in the subgroup of patients treated with adjuvant chemotherapy (HR 1.44; p = 0.381 and HR 2.48; p = 0.003, respectively). In contrast, the bad prognostic impact of high uPA and PAI-1 levels was lost in the subgroup of patients treated with adjuvant hormone therapy (HR 0.79; p = 0.693 and HR 0.26; p = 0.204, respectively). The same observations were made for the uPA/PAI-1 combination. Our study confirmed the prognostic value of serine proteases in early breast cancer. In addition, it pointed to a possible predictive value of these tumor markers for response to adjuvant hormone therapy with tamoxifen, which should be confirmed in further studies.
Urokinase-type plasminogen activator (uPA), its inhibitors (PAI-1 and PAI-2), and its receptor (uPAR) play a key role in tumor invasion and metastasis. This study was designed to evaluate the ...prognostic impact of uPA, PAI-1, PAI-2, and uPAR and the combination of these factors in a group of 460 primary breast cancer patients. Concentrations of all 4 components of the uPA system were measured in tumor extracts using enzyme-linked immunosorbent assays (American Diagnostica, Inc, Greenwich, CT). After a median follow-up of 33 months, 18.5% of the patients had relapsed. The Cox proportional hazards model was applied for both univariate and multivariate analyses of disease-free survival (DFS). PAI-1 and PAI-2 were shown to provide independent prognostic information in breast cancer. Patients with either low levels of PAI-1 or high levels of PAI-2 were found to have better DFS (relative risk was 2.08 and 1.78, respectively). The prognostic value could be even further improved by a combination of both inhibitors. Aside from the uPA inhibitors, only nodal status and hormonal receptor status retained independent prognostic value. The other 2 invasion markers, uPA and uPAR, showed no statistically significant impact on DFS. In our patients, who were mostly treated with adjuvant therapy, uPA was not found to be an independent prognostic marker for DFS; this could be a consequence of the predictive value of uPA for response to adjuvant therapy and should be further investigated.
The aim of this study was to determine urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type 1 (PAI-1) concentrations in tumour and adjacent normal tissue samples from ...58 patients, and in serum samples from 40 of 58 patients with squamous cell carcinoma of the head and neck obtained at diagnosis and after completion of therapy. uPA and PAI-1 serum concentrations were also measured in 28 healthy volunteers who served as controls. Measurements were made using enzyme-linked immunosorbent assay (ELISA) techniques. For both uPA and PAI-1, significantly elevated concentrations were measured in tumour tissue as compared with normal tissue (uPA: 8.89 versus 0.41
ng/mg total protein (mgp),
P<0.0001; PAI-1: 23.9 versus 1.47
ng/mgp,
P<0.0001). A statistically significant difference in uPA concentrations was found between normal laryngeal and non-laryngeal tissue (0.52 versus 0.3
ng/mgp,
P=0.008), and in PAI-1 concentrations between T
1+2 and T
3+4 stage of disease (17.32 versus 35.63
ng/mgp,
P=0.04). The uPA concentrations positively correlated with those of PAI-1 measured in both tumour (
R
S=0.62,
P<0.0001) and normal tissue (
R
S=0.30,
P=0.02). In serum samples, lower concentrations of PAI-1 were measured in the control group than in patients with cancer (412.0 versus 680.5
ng/ml serum (mls),
P=0.0006). The time of collection of the serum sample did not influence uPA and PAI-1 concentrations, and no association was observed between their concentrations and any clinical and histopathological prognostic factors tested. Our results indicate that both uPA and PAI-1 may play a specific role in the process of invasion and metastasis, and might also be of prognostic value in squamous cell carcinoma of the head and neck.
The aim of the study was to evaluate the prognostic significance of tumour and serum concentrations of urokinase-type plasminogen activator (uPA), its type 1 inhibitor (PAI-1) and cathepsin D (Cath ...D) in patients with squamous cell carcinoma of the head and neck (SCCHN).
Determinations of uPA and PAI-1 were made using enzyme-linked immunosorbent assays in tumour and serum samples of 47 and 32/47 patients, respectively. For the determination of tumour (94 patients) and serum (34/94 patients) Cath D concentrations, an immunoradiometric assay was used.
In an univariate survival analysis, the risk of disease recurrence and SCCHN-related death was significantly higher in the patients with high uPA (P = 0.046, P = 0.010) tumours, compared to those with low uPA tumours. In addition, the high serum levels of uPA correlated positively with the rate of relapse (P = 0.007), but not with the mortality rate (P = 0.200). There was no statistically significant difference between low and high PAI-1 groups, regarding either tumour or serum concentration of the inhibitor, and between low and high Cath D tumours. Low Cath D serum levels appeared to be related to longer disease-free interval (P = 0.055), but not to disease-specific survival (P = 0.120).
The tumour levels of uPA, as well as serum levels of uPA and Cath D could potentially predict the survival probability of patients with SCCHN. However, the strength of this association remains to be investigated on a larger and more homogeneous group of patients.
The independent prognostic value of protease uPA and its inhibitor PAI-1 for survival in breast cancer patients is firmly established. However, there is very little data on the prognostic value of ...serine proteases and their inhibitors for locoregional recurrence in breast cancer. The prognostic value of PAI-1 for local control in a group of 766 patients treated at our institute with either breast conserving treatment or modified radical mastectomy was evaluated. The locoregional recurrence-free survival (LRFS) of patients with PAI-1 values above the median value was significantly worse than that of patients with PAI-1 values below the median value (log-rank; p=0.0078). In multivariate analysis PAI-1 levels proved to be of independent statistical significance for LRFS (p=0.0401, relative risk 2.28, 95% confidence interval 1.04-5.02). The independent prognostic value of PAI-1 for metastasis-free survival and overall survival was also confirmed. In addition, our data suggest that PAI-1 antigen levels in tumor tissue might be of prognostic value for survival after locoregional recurrence (log-rank; p=0.0618). According to our findings, PAI-1 levels could be used as a biological marker that could facilitate the identifation of patients with a higher risk of local relapse already at the time of primary treatment. These patients should then be offered more aggressive treatment.
The association between drug-resistance and three markers for invasive capacity: cathepsin D (Cath D), urokinase type plasminogen activator (uPA) and inhibitor of plasminogen activator type 1 (PAI-1) ...was examined in nine cervical and laryngeal carcinoma cell lines resistant to different cytostatics. The level of Cath D was measured by solid phase two-site immunoradiometric assay, while uPA and PAI-1 concentrations were determined by use of ELISA. All drug resistant cell lines had increased concentration of cathepsin D. uPA levels were similar in parental and drug resistant cervical carcinoma cells, but significantly higher in all examined drug resistant laryngeal carcinoma cells. In cervical carcinoma cells, PAI-1 concentrations were similar in parental and cisplatin resistant, but significantly higher in doxorubicin resistant cells. In laryngeal carcinoma cells, no increase in concentrations of PAI-1 was determined in the three from five resistant cell lines. There was no uPA in conditioned medium of parental or drug resistant cells. PAI-1 was detected in conditioned medium. Its levels were significantly increased in the medium of two cervical and three laryngeal drug resistant carcinoma cells. Thus, our results suggest that drug-resistance may be accompanied by increased levels of tumor associated proteases and/or its inhibitor.
The clinical determination of proteases which are involved in carcinogenesis, invasion and metastasis may contribute to the detection of the early stage of disease, and to the prognostic assessment ...of patients with the cancer. The aim of the present study was to determine the level of urokinase plasminogen activator (uPA), plasminogen activator inhibitor type 1 (PAI-1) and plasminogen activator inhibitor type 2 (PAI-2) in normal and malignant tissues of corpus uteri and to evaluate the possible correlation with clinical and histopathological prognostic factors. UPA, PA-I and PAI-2 were determined by the ELISA assay in tissue cytosol of matched pair samples from 27 patients with endometrial carcinoma. Results show that significantly higher levels of these proteins were found in malignant than in normal tissue samples (uPA: 1.266 versus 0.633 ng/mg protein, PAI-1:4.468 versus 1.958 ng/mg protein, and PAI-2:3.428 versus 0.483 ng/ml protein). The levels of uPA and PAI-1 did not correlate with clinical staging or pathohistological grading. However, in tumor tissues with clinical stages II and III, myometrial invasion > 50%, and lymphovascular invasion, increased levels of PAI-2 were determined. Our results indicate that components of the plasminogen activation cascade are up-regulated in endometrial cancer and suggest the role of PAI-2 in determining invasive potential of endometrial carcinomas.
Cysteine
proteinases cathepsins (Cats) B and L and their endogenous inhibitors
stefins (Stefs) A and B are implicated in the processes of local and
metastatic tumor spread. They were identified as ...potential
prognosticators in various malignant diseases, particularly in breast
cancer. The aim of the present study was to determine the
concentrations of Cats B and L and Stefs A and B in the tumor and
adjacent normal tissue samples collected from 49 patients (the present
group) with squamous cell carcinoma of the head and neck (SCCHN), using
quantitative immunosorbent assays (ELISA; KRKA d.d., Novo mesto,
Slovenia). Their clinical significance was compared with that from a
previous study (the reference group, 45 patients; Budihna et
al. , Biol. Chem. Hoppe-Seyler, 377: 385–390,
1996). The follow-up of patients from the latter report was updated for
this purpose.
In the present group, significantly higher concentrations of Cat B
( P < 0.0001), Cat L ( P <
0.0001) and Stef A ( P = 0.006) were found in tumors
compared with concentrations in their normal tissue counterparts. Cat
concentrations in normal laryngeal tissue were significantly/marginally
elevated compared with nonlaryngeal tissue (Cat B,
P = 0.02; Cat L, P = 0.06). The
tumor concentration of Cat L was found to correlate with pT
classification ( P = 0.005) and
tumor-node-metastasis stage ( P = 0.05),
whereas the concentrations of Stefs A and B correlated with pN
classification ( P = 0.007 and P = 0.03, respectively) and tumor-node-metastasis stage of the
disease ( P = 0.02 and P = 0.03,
respectively). There was no statistically significant difference
between low and high Cat B or Cat L groups, regarding either
disease-free survival or disease-specific survival, using a
minimum P approach to determine cutoff concentrations.
The risk of disease recurrence and SCCHN-related death was
significantly higher in patients with low Stef A ( P = 0.0006 and P = 0.0005, respectively) and Stef B
( P = 0.0009 and P = 0.0007,
respectively) tumors, compared with those with high-Stef A and Stef B
tumors. These results remained significant even after P s
were adjusted for a possible bias in the estimated effect on survival.
The survival analysis in the reference group also confirmed these
findings (Stef A: P = 0.0009 and
P = 0.002, respectively; Stef B:
P = 0.03 and P = 0.009,
respectively). To avoid any possible bias arising from the differences
between the laboratories that performed the biochemical analysis, the
concentrations of both Stefs in the present group and in the reference
group were standardized and coupled together to form a uniform group.
In univariate survival analysis, standardized values of Stef A and Stef
B correlated inversely with the rate of relapse ( P = 0.0000) and mortality rate ( P = 0.0000).
Multivariate regression analysis showed that the standardized value of
Stef A is the strongest independent prognostic factor for both
disease-free survival and disease-specific survival. These findings
show the specific role of Cats B and L and Stefs A and B in the
invasive behavior of SCCHN. Furthermore, Stef A proved to be a reliable
prognosticator of the risk of relapse and death in patients with this
type of cancer.
