Advertisement

Plasma levels of E-cadherin and MMP-13 in prostate cancer patients: correlation with PSA, testosterone and pathological parameters

Abstract

Aims and Background

Prostate cancer is the most common malignant tumor in men. Serum prostate-specific antigen (PSA), Gleason score and clinical range at the time of diagnosis are important factors to predict prognosis and outcome after therapy but additional accurate and reliable biomarkers are still wanted. So far, few biomarkers of prostate cancer have been successfully implemented and are being used in clinical practice. However, modifications of E-cadherin and MMP-13 expression are likely to be involved in prostate cancer invasion and thus are potential biomarkers for prognosis.

Patients and Methods

We analyzed the concentrations of E-cadherin and MMP-13 in plasma of patients with prostate cancer at diagnosis and 3 and 6 months after the beginning of treatment and related these measures to free and total PSA and other clinical features.

Results

The concentration of E-cadherin was lower in patients with prostate cancer compared to the control group, but there was no difference in the concentration of MMP-13 between these two groups. During treatment, however, we found no significant differences between the concentrations of MMP-13 and E-cadherin, but we observed a significant positive correlation between total PSA and E-cadherin plasma concentration at the third month of treatment and between total testosterone and MMP-13 plasma concentration before the start of treatment.

Conclusions

The results suggest that these parameters could be used both in the diagnosis and prognosis of prostate cancer.

Tumori 2015; 101(2): 185 - 188

Article Type: ORIGINAL RESEARCH ARTICLE

DOI:10.5301/tj.5000237

Authors

Carolina M. Bonaldi, Ligia A. Azzalis, Virginia B.C. Junqueira, Claudia G.B. de Oliveira, Viviane A. Vilas Boas, Thaís M. Gáscon, Flávia S. Gehrke, Renata K. Kuniyoshi, Beatriz C.A. Alves, Fernando L.A. Fonseca

Article History

Disclosures

Financial support: This work was funded by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP: 10/50490-6). FSG and BCAA were supported by a CAPES postdoctoral fellowship; RKK was supported by a UNIEMP doctoral fellowship.
Conflict of interest: The authors have no conflict of interest.

This article is available as full text PDF.

Download any of the following attachments:

Introduction

Prostate cancer (PC) is the sixth most common cancer type in the world. As in other tumors, malignant transformation of prostate cells is a result of the accumulation of genetic mutations. The most widely used marker for PC detection in the clinic is serum prostate-specific antigen (PSA), whose ability to distinguish PC from benign prostate hyperplasia is improved by the free-to-total PSA ratio in patients with PSA between 4 and 10 ng/mL (1). However, the aggressiveness and metastatic potential of PC are difficult to predict based only on PSA screening, Gleason score and clinical parameters, which makes the use of novel molecular parameters for these traits highly necessary.

E-cadherin belongs to a class of type-1 transmembrane proteins that play important roles in cell adhesion and communication. Multiple studies have found reduced E-cadherin expression in aggressive PC, suggesting it might be a metastasis suppressor (2). Matrix metalloproteinases (MMPs), in turn, are a family of enzymes involved in the breakdown of the extracellular matrix in normal physiological processes. Normal tissue structure is a result of a balanced interaction between MMPs and the tissue inhibitors of metalloproteinases (TIMPs). These enzymes play an essential role in tumor invasion and metastasis, and MMP-13 is the one most commonly found in carcinomas (3). MMPs and their TIMPs are generally correlated with tumor differentiation.

Since modifications of E-cadherin and MMP-13 expression are likely to be involved in PC invasion, we analyzed their concentrations in plasma of patients with PC at diagnosis and during treatment, and related their concentrations to free and total PSA as well as other clinical features in order to ascertain if these measures could be of clinical help in establishing the metastatic risk in patients with PC.

Patients and Methods

This study was carried out in accordance with the ethical principles and was approved by the local ethics committee. After having obtained the patients’ informed consent, we collected about 15.0 mL of peripheral blood from 29 patients with PC treated at the Urology Clinic of the Faculty of Medicine of ABC for plasma evaluation of E-cadherin, MMP-13, free and total PSA, and testosterone at diagnosis and after 3 and 6 months of treatment. The follow-up schedule also included other clinical examinations. Only patients with prostate adenocarcinoma without any previous treatment were included in the study; patients were excluded if they had benign prostatic hyperplasia, HIV or hepatitis B or C infection, or autoimmune diseases. The control group of healthy males consisted of 10 men with total PSA ≤1.5 ng/mL and normal digital rectal examination; the men in the control group had the same average age as the patients.

Plasma concentrations of E-cadherin and MMP-13 were determined using enzyme immunoassay (R&D Systems, Inc and IBL International, respectively). PSA and testosterone were measured using the commercially available Siemens 06607631 Immulite® testosterone kit and Siemens 06607968 Immulite® third-generation PSA kit, respectively. Histological grade according to the Gleason score was assessed on stained sections in accordance with the WHO International Classification IARC/WHO Classification of Tumours. Histologically, all tumors were adenocarcinomas.

For variables of approximately normal distribution (patients’ age), Student’s t-test was used to compare the mean values of independent variables. In other cases (biomarkers), differences between groups were analyzed with the Mann-Whitney U-test. Correlations between pairs of parameters were analyzed using Spearman’s correlation coefficient. To predict the grade of tumor, receiver-operating characteristics (ROC) curves were generated. A p value <0.05 was accepted as the limit of statistical significance. The calculations were performed using the commercial SPSS software package.

Results and Discussion

The clinical characteristics of patients and controls are summarized in the supplemental material (Supplementary Tables S-1 and S-II, respectively. Available online at www.tumorijournal.com).

At diagnosis, the E-cadherin concentration was lower in patients with PC than in the control group (p<0.001) (Fig. 1). Decreased expression of E-cadherin in patients with PC was also described previously (4, 5), and this finding was associated with a worse prognosis. However, as can be seen in Figure 1, there was no difference in the MMP-13 concentration between the 2 groups (p = 0.2085). Szarvas et al (6) also found no significant difference in MMP-13 expression between PC patients and healthy men, despite the known involvement of MMPs in PC. Fernandez-Gomez et al (7), using tissue microarray to evaluate 7 different types of metalloproteinase, showed a significant association between increased expression of MMP-13 and elevated levels of PSA. It is clear, thus, that different methodologies could be a limiting factor in associating MMP-13 with PSA in patients with PC, since the serological results described here did not match those obtained by tissue microarray (Fig. 1).

Comparison of MMP-13 and E-cadherin (E-CAD) plasma concentrations (ng/mL) between controls and patients with prostate cancer.

Logistic regression was performed to determine the parameters that could be used as prognostic factors for PC (Tab. I and Fig. 2).

Logistic regression to determine which parameters could be used as prognostic factors for prostate cancer

Parameters Odds ratio P value
E-cadherin - 0.2658
MMP-13 - 0.1772

Total PSA versus MMP-13 (A) and E-cadherin (B) ROC curves. Red lines represent reference; blue lines, total PSA; green lines, MMP-13 (A) or E-cadherin (B). The areas obtained from the ROC curves were 0.97 for total PSA (p<0.001), 1.00 for E-cadherin (p<0.001), and 0.64 for MMP-13 (p = 0.204).

During treatment, however, using the nonparametric Friedman test, we found no significant differences between the concentrations of MMP-13 (p = 0.966) and E-cadherin (p = 0.166) at diagnosis and after the third and sixth months of treatment (p = 0.786). However, we observed a significant positive correlation (Spearman’s correlation coefficient) between total PSA and E-cadherin plasma concentration at the end of the third month of treatment (Tab. II). Our results are in agreement with those reported by Gravdal et al (4), although these authors did not indicate the times of measurement during treatment.

Spearman’s correlation coefficient for total and free PSA, E-cadherin, and MMP-13 at 0, 3 and 6 months of treatment

Follow-up E-CAD MMP-13
E-CAD = E-cadherin; MMP-13 = matrix metalloproteinase 13; TPSA = total PSA; FPSA = free PSA; mo = months.
TPSA r 0.129 -0.201
p 0.504 0.296
0
FPSA r 0.111 -0.131
p 0.567 0.500
TPSA r 0.393 0.050
p 0.035 0.797
3 mo
FPSA r 0.353 0.037
p 0.060 0.849
TPSA r 0.161 0.152
p 0.405 0.431
6 mo
FPSA r 0.167 -0.191
p 0.387 0.320

Spearman’s correlation coefficient also showed a significant positive correlation between total testosterone and MMP-13 plasma concentration before the start of treatment. On the other hand, a negative correlation was observed after the sixth month of treatment (Tab. III). The E-cadherin plasma concentration showed a negative correlation with total testosterone after the third month of treatment and with free testosterone after the third and sixth months of treatment.

Spearman’s correlation coefficient between total and free testosterone, MMP-13 and E-cadherin plasma concentrations at 0, 3 and 6 months of treatment

Follow-up TTesto FTesto
E-CAD = E-cadherin; MMP-13 = matrix metalloproteinase 13; TTesto = total testosterone; FTesto = free testosterone; mo = months.
MMP-13 r 0.433 0.249
p 0.019 0.193
0
E-CAD r -0.215 -0.041
p 0.263 0.833
MMP-13 r -0.086 0.024
p 0.659 0.902
3 mo
E-CAD r -0.434 -0.640
p 0.019 <0.001
MMP-13 r -0.384 0.064
p 0.040 0.740
6 mo
E-CAD r -0.082 -0.468
p 0.671 0.010

At the initial stages, PC is dependent on androgens for growth. However, prostate tumors eventually recur due to the transition from an androgen-dependent to an androgen-independent state, leading to highly metastatic disease for which there is no effective therapy available (8). Interference in tumor biology promoted by treatment in patients with PC has been previously reported by our group (9, 10). It is known that increased circulating E-cadherin levels may have different causes; however, the involvement of testosterone in this process is still unknown, despite reports in the literature (11).

Our results on the E-cadherin plasma concentration suggest that testosterone and PC markers were associated with tumor progression to metastasis. In 2006, Buhmeida et al (12) found that reduced levels of E-cadherin were correlated with advanced Gleason score (p = 0.003). However, these results were obtained with immunohistochemistry, while the E-cadherin and MMP-13 concentrations in our study were measured in plasma.

There was no difference between E-cadherin and MMP-13 plasma levels in the group with perineural invasion.

Conclusions

The concentration of E-cadherin was lower in patients with PC compared to the control group, but there was no difference in the concentration of MMP-13 between the 2 groups. These results suggest that these parameters could be used both in the diagnosis and prognosis of PC.

Disclosures

Financial support: This work was funded by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP: 10/50490-6). FSG and BCAA were supported by a CAPES postdoctoral fellowship; RKK was supported by a UNIEMP doctoral fellowship.
Conflict of interest: The authors have no conflict of interest.
References
  • 1. Masieri L.,Minervini A.,Vittori G. The role of free to total PSA ratio in prediction of extracapsular tumor extension and biochemical recurrence after radical prostatectomy in patients with PSA between 4 and 10 ng/ml. Int Urol Nephrol 2012; 44: 1031-1038 Google Scholar
  • 2. Khamis ZI.,Iczkowski KA.,Sang Q-XA. Metastasis suppressors in human benign prostate, intraepithelial neoplasia, and invasive cancer: their prospects as therapeutic agents. Med Res Rev 2012; 32: 1026-1077 Google Scholar
  • 3. Morgia G.,Falsaperla M.,Malaponte G. Matrix metalloproteinases as diagnostic (MMP-13) and prognostic (MMP-2, MMP-9) markers of prostate cancer. Urol Res 2005; 33: 44-50 Google Scholar
  • 4. Gravdal K.,Halvorsen OJ.,Haukaas SA.,Akslen LA. A switch from E-cadherin to N-cadherin expression indicates epithelial to mesenchymal transition and is of strong and independent importance for the progress of prostate cancer. Clin Cancer Res 2007; 13: 7003-7011 Google Scholar
  • 5. Jaggi M.,Johansson SL.,Baker JJ.,Smith LM.,Galich A.,Balaji KC. Aberrant expression of E-cadherin and beta-catenin in human prostate cancer. Urol Oncol 2005; 23: 402-406 Google Scholar
  • 6. Szarvas T.,Becker M.,Vom Dorp F. Elevated serum matrix metalloproteinase 7 levels predict poor prognosis after radical prostatectomy. Int J Cancer 2011; 128: 1486-1492 Google Scholar
  • 7. Fernandez-Gomez J.,Escaf S.,Gonzalez LO. Relationship between metalloprotease expression in tumour and stromal cells and aggressive behaviour in prostate carcinoma: Simultaneous high-throughput study of multiple metalloproteases and their inhibitors using tissue array analysis of radical prostatectomy samples. Scand J Urol Nephrol 2011; 45: 171-176 Google Scholar
  • 8. Zhu M-L.,Kyprianou N. Role of androgens and the androgen receptor in epithelial-mesenchymal transition and invasion of prostate cancer cells. FASEB J 2010; 24: 769-777 Google Scholar
  • 9. Delgado PO.,Alves BC.,Gehrke FS. Characterization of cell-free circulating DNA in plasma in patients with prostate cancer. Tumour Biol 2013; 34: 983-986 Google Scholar
  • 10. Wroclawski ML.,Serpa-Neto A.,Fonseca FL. Cell-free plasma DNA as biochemical biomarker for the diagnosis and follow-up of prostate cancer patients. Tumour Biol 2013; 34: 2921-2927 Google Scholar
  • 11. Roberts PC.,Mottillo EP.,Baxa AC. Sequential molecular and cellular events during neoplastic progression: a mouse syngeneic ovarian cancer model. Neoplasia 2005; 7: 944-956 Google Scholar
  • 12. Buhmeida A.,Pyrhönen S.,Laato M.,Collan Y. Prognostic factors in prostate cancer. Diagn Pathol 2006; 1: 4- Google Scholar

Authors

Affiliations

  • Department of Urology, ABC Medical School, Santo André, SP - Brazil
  • Biological Science Department, UNIFESP, Diadema, SP - Brazil

Article usage statistics

The blue line displays unique views in the time frame indicated.
The yellow line displays unique downloads.
Views and downloads are counted only once per session.

This article has supplementary materials available to download.