Expression of E26 transformation specific-1 (ETS-1) in tumour-infiltrating lymphocytes (TILs) is adverse prognostic factor in invasive breast cancer.

AIM OF THE STUDY: The microenvironment depicts the relationship between tumour cells and immune response, and every insight into stromal lymphocytes could contribute to explain their role and activity. E26 transformation specific-1 (ETS-1) is a transcription factor that is active in cell proliferation. We analysed its immunohistochemical expression in tumour infiltrating lymphocytes (TILs) in invasive breast cancer and correlated its immunohistochemical score (IHS) to traditional predictive and prognostic factors and survival. MATERIALS AND METHODS: The sample contains data of 121 patients with invasive breast cancer, not otherwise specified (NOS) who underwent mammectomy and lymphadenectomy in 2002 at the Clinical Hospital Centre Zagreb, Croatia. Paraffin blocks of the tumour tissue were collected from the pathological archive. Three representative areas of every patient were chosen and multiple tissue samples were made. Immunohistochemical staining with rabbit anti-ETS-1 (Novocastra, UK) and the ABC method was performed on a DAKO Autostainer. The expression of ETS-1 in stromal TILs was analysed on an Olympus 41 microscope. The IHS score was calculated and correlated with clinical and pathological parameters, as well as disease-free survival (DFS) and overall survival (OS). RESULTS: In almost all patients (95%), some expression of ETS-1 in TILs was found. A moderate/high score of ETS-1 correlated with larger tumour size and higher histological grade, high proliferation index and low progesterone receptors (PgR). The patients with moderate/high ETS-1 expression in TILs had shorter DFS than patients with weak/negative ETS-1 expression. CONCLUSION: In invasive breast cancer NOS, expression of ETS-1 in TILs is an adverse prognostic factor.

Microsatellite instability in metaplasia-dysplasia-adenocarcinoma sequence of Barrett esophagus: a retrospective study.

AIM: To analyze the loss of mismatch repair (MMR) system protein expression in metaplasia-dysplasia-adenocarcinoma sequence of Barrett esophagus (BE). METHODS: This study retrospectively analyzed the data from 70 patients with pathohistological diagnosis of BE or esophageal adenocarcinoma (EAC) treated at the Clinical Department of Pathology and Cytology, University Hospital Center Zagreb, from January 2009 to January 2011. Patients were divided into three groups: BE without dysplasia (22 patients), BE with dysplasia (37 patients), and EAC (11 patients). Immunohistochemical expression of MutL homologue 1 (MLH1), MutS homologue 2 (MSH2), postmeiotic segregation increased 2 (PMS2), and MutS homologue 6 (MSH6) of DNA MMR system was measured and compared with tumor protein p53 expression. RESULTS: A total of 81.8% and 81.8% patients with EAC, 32.4% and 35.1% patients with dysplasia, and 50% and 54.5% patients without dysplasia had loss of MLH1 and PMS2 expression, respectively. Patients with EAC and patients with dysplasia did not have loss of MSH2 and MSH6 expression, and 18.2% patients without dysplasia had loss of MSH2 and MSH6 expression. There was a strong positive correlation between MLH1 and PMS2 expression (Spearman ρ 0.97; P<0.001) and between MSH2 and MSH6 expression (Spearman ρ 0.90, P<0.001) in the entire sample and in all BE groups. No significant correlations of MLH1 and PMS2 with p53 expression were found, except in dysplasia group (φ 0.402, P=0.030 for MSH1; φ 0.371, P=0.042 for PMS2). CONCLUSION: Although we demonstrated considerable loss of MLH1 and PMS2 expression in BE-associated carcinoma sequence, due to the retrospective study design and low number of patients we cannot conclude that MLH1 and PMS2 can be used as biomarkers for patient surveillance and therapy-making decisions. Oxford Centre for Evidence-based Medicine level of evidence: 3.

Dipeptidyl peptidase 9 (DPP9) in human skin cells.

BACKGROUND: Dipeptidyl peptidase 9 (DPP9) is a relatively new member of the DPPIV family of prolyl dipeptidases which is ubiquitously expressed. Its role in regulation of immune responses and proliferation of epithelial carcinoma cells was reported. There is no data on possible role of DPP9 expressed in skin epithelial cells (keratinocytes) and in dermal fibroblasts. MATERIALS AND METHODS: Transcriptional and protein expression of DPP9 and DPPIV was examined in fibroblasts and keratinocytes isolated from normal human skin. Localization of DPP9 and its sub-localization in Golgi were determined by immunocytochemistry staining. DPPIV-like enzyme activity was determined in cell lysates and in isolated cell fractions containing membranes (M), cytosol (C) and content of organelles/endosomes/vesicles (V). Relative contribution of DPPIV and DPP8/9 enzyme activity in these fractions was determined by using selective inhibitors: sitagliptin (selective for DPPIV) and 1G244 (selective for DPP9 and a highly homologous DPP8). Possible roles of DPP8/9 via its enzyme activity were analysed by assessment of survival and proliferative capacity of fibroblasts and HaCaT cells of keratinocyte origin in the presence of the inhibitors. Possible role of DPP9 in cell migration and/or adhesion was analysed in fibroblasts and HaCaT cells after DPP9 gene silencing. RESULTS: Fibroblasts and keratinocytes exerted comparable level of DPP9 both at transcriptional and protein level. Fibroblasts strongly expressed DPPIV, whereas in keratinocytes DPPIV expression was low. DPP9 expression was found in cytosol and in perinuclear area of some fibroblasts, or in scattered pattern of keratinocytes, as well as in nuclei of some cells. Only low level of DPP9 sub-localization within Golgi was observed in fibroblasts and keratinocytes. DPPIV-like enzyme activity was about 5 times higher in lysates of fibroblasts than of HaCaT cells. In fibroblasts DPPIV-like enzyme activity was mainly (65%) found in the fraction containing cell membranes (M) and was predominantly (86.9%) due to DPPIV. In contrast, in HaCaT cells the DPPIV-like enzyme activity was mainly (84.2%) found in cytosol (C) and was predominantly (95.6%) due to DPP8/9. Survival and the proliferative capacity were significantly diminished in the presence of 10µM 1G244, both in fibroblasts and in HaCaT cells, suggesting possible role of DPP8/9 enzyme activity in regulation of survival and proliferation of these cells. DPP9 gene silencing resulted in decreased adhesion of fibroblasts, as well as in decreased migration of fibroblasts and HaCaT cells. Accumulation of DPP9 on the edges of plasma membranes of fibroblasts and keratinocytes adhering to surface supports the idea of possible role of DPP9 in cell adhesion. CONCLUSIONS: This is the first study showing protein expression, sub-localization and possible biological roles of DPP9 expressed in isolated human skin cells. The data may be relevant for development of new drugs against skin diseases by targeting DPP9 expressed in the skin cells.

Telomerase activity in breast cancer patients: association with poor prognosis and more aggressive phenotype.

Telomerase expression is an important mechanism of tumor unlimited replicative potential. The aim of this study was to evaluate prognostic impact of telomerase activity in breast cancer patients and to correlate telomerase activity with established prognostic factors. We analyzed tissue of 102 malignant breast lesions and 20 healthy breast tissues. Telomerase activity was determined by telomeric repeat amplification protocol assay. Telomerase activity was present in 77 (75.49 %) of 102 breast cancers. Telomerase activity in breast cancers was statistically significantly higher in comparison with the activity in normal breast tissue. The levels of telomerase activity were significantly positively correlated with tumor size, axillary nodal status, histological grade, HER-2/neu protein expression in tumor tissue and expression of the nuclear antigen Ki-67. A statistically significant negative correlation was found between the presence of ER and telomerase activity. There was no correlation between telomerase activity and concentration of PR or the age of patients. Kaplan-Meier analysis showed that patients with higher telomerase activity had significantly shorter 10-year disease-free survival (p < 0.0001) and 10-year overall survival (p < 0.0001) than those with lower telomerase activity. These results were confirmed by logistic regression analysis. Our results support the prognostic role of telomerase activity and its relationship with the more aggressive phenotype of breast cancer.

Role of breast imaging in predicting outcome of lesions of uncertain malignant potential (B3) diagnosed at core needle biopsy.

AIMS: To elucidate whether breast imaging can predict final histologic diagnosis of lesions of uncertain malignant potential diagnosed at ultrasound core needle biopsy (CNB). METHODS: The imaging characteristics (mammography, ultrasound, and magnetic resonance imaging [MRI]) of lesions of uncertain malignant potential in the breast that were obtained by ultrasound CNB were retrospectively analyzed in 87 women. Radiologic characteristics of lesions were compared to definitive histopathologic findings. RESULTS: Out of 87 breast lesions of uncertain malignant potential, 27 (31%) were diagnosed as papillary lesions, 24 (28%) atypical ductal hyperplasia, 19 (22%) lobular intraepithelial neoplasia, 9 (10%) phyllodes tumors, 3 (3%) radial sclerosing lesions, and 5 (6%) unspecified lesions of uncertain malignant potential. The underestimation rate of malignancy at CNB based on the total number of lesions on final follow-up was 22%. Using multivariate logistic regression, Breast Imaging-Reporting and Data System (BI-RADS) score (odds ratio [OR] = 12.29, p = 0.027) and Göttingen MRI scoring system (OR = 8.1, p = 0.008) were found to be independent predictors of malignancy. Receiver operating characteristic analysis showed that Göttingen MRI score >3 provides a plausibly good cutoff value with sensitivity of 100 (95% confidence interval [CI] 74%-100%) and specificity of 76% (95% CI 61%-88%). CONCLUSIONS: Lesions of uncertain malignant potential classified as BI-RADS 5 and Göttingen score 4 or higher are at significantly higher risk of harboring malignancy and therefore should be recommended for surgical excision.

[CLINICAL GUIDELINES FOR DIAGNOSIS, TREATMENT AND MONITORING OF PATIENTS WITH INVASIVE BREAST CANCER--CROATIAN ONCOLOGY SOCIETY]. / KLINICKE UPUTE HRVATSKOGA ONKOLOSKOG DRUSTVA ZA DIJAGNOZU, LIJECENJE I PRACENJE BOLESNICA/KA OBOLJELIH OD INVAZIVNOG RAKA DOJKE.

Breast cancer is the most common cancer in women. It can be diagnosed in early stage through screening, early detection and educational programs, and when diagnosed early it can be efficiently treated. Treatment modalities include surgery, chemotherapy, radiotherapy, hormonal therapy and targeted biologic therapy, according to the stage of the disease and patient condition. Treatment decisions should be made after multidisciplinary team discussion. Due to the significance of this disease it is important to define and implement standardized approach for diagnostic, treatment and monitoring algorithm as well. The following text presents the clinical guidelines in order to standardize the procedures and criteria for diagnosis, management, treatment and monitoring of patients with breast cancer in the Republic of Croatia.

[COLONIC MUCO-SUBMUCOSAL ELONGATED POLYP: A CASE REPORT AND LITERATURE REVIEW]. / IZDUZENI SLUZNICNO-PODSLUZNICNI POLIP DEBELOG CRIJEVA: PRIKAZ BOLESNIKA I PREGLED LITERATURE.

Recently a new entity has been described--a colonic muco-submucosal elongated polyp (CMSEP)--that did not fall into traditional classification of colorectal polyps. The CMSEP is endoscopically characterised by elongated, worm-like appearance with a normal overlying mucosa. Histologic characteristics of the CMSEP comprise mucosa and expanded submucosa with dilated vasculature and lymphatics. Herein, we report a case of CMSEP, that to the best of our knowledge, has not been previously described in our literature. With regard to the on-going National colorectal cancer screening programme, our intention is to draw attention of gastrointestinal pathologists and endoscopists to this distinctive and very rare phenomenon.

Immunohistochemical expression of 8-oxo-7,8-dihydro-2'-deoxyguanosine in cytoplasm of tumour and adjacent normal mucosa cells in patients with colorectal cancer.

BACKGROUND: The aim of this research was to study the levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) in tumour tissue samples of colorectal carcinoma based upon immunohistochemical detection and compare those results with patients' outcome. METHODS: Tumour blocks of patients surgically treated for colorectal cancer were evaluated by 8-oxodG immunohistochemical staining. The expression was analysed in 500 tumour cells. The percentage of positive cells, as well as staining intensity, was recorded, and Allred score was calculated. For each patient, data of age, gender, tumour size and location, margin status, histologic grade, tumour stage, lymph node status, vascular invasion, overall survival, and therapy protocols were collected. Tumour grade was divided into two groups as low and high grade. RESULTS: In this study, 146 consecutive patients with primary colorectal carcinoma were included. All data were available for 138 patients, and they were included in this research. There were 83 male and 55 female patients; the median age was 64 years (range 35-87 years). The results showed shorter 5- and 10-year survival in patients with 8-oxodG positive tumour cells (5-year survival, n=138, Mantel-Cox, chi-square 4.116, degree of freedom (df)=1, p<0.05; 10-year survival, n=134, Mantel-Cox, chi-square 4.374, df=1, p<0.05). The results showed a positive correlation between Allred score and high tumour grade (two-tailed Spearman's ρ 0.184; p<0.05), as well as with non-polypoid tumour growth (two-tailed Spearman's ρ 0.198; p<0.05). There was no significant difference of 8-oxodG expression related to age, sex, blood group, size and tumour site, distance from the edge of the resected tumour margin, lymph nodes involvement, and vascular invasion. CONCLUSIONS: In this study, the positive correlation between 8-oxodG presence in the tumour cells, worse clinical outcome, higher tumour grade, and flat morphology was found.

[Clinical recommendations for diagnosis, treatment and monitoring of patients with cancer of unknown primary site]. / Klinicke preporuke za dijagnozu, lijecenje i pracenje bolesnika oboljelih od raka nepoznata primarnog podrijetla.

Cancer of unknown primary (CUP) site comprises very heterogeneous group of various malignant tumors presented in metastatic phase of the disease. Diagnosis is set when primary site remains unidentified after a thorough diagnostic evaluation in patients with histologically proven malignant metastatic disease. Despite poor prognosis in most patients, favorable prognostic clinical entities have been recognized constituting the most important group of patients for oncological treatment. The following text presents the clinical guidelines in order to standardize the diagnosis, treatment and follow-up of patients with cancer of unknown primary site in the Republic of Croatia.

Breast Cancer Molecular Subtypes and Oxidative DNA Damage.

BACKGROUND: Oxidative stress is thought to play a major role in etiology of many cancers, including breast cancer. 8-hydroxy-2'deoxyguanosine (8-OHdG) is the most abundant marker of oxidative DNA damage. The aim of this study was to assess the extent of oxidative DNA damage in different breast cancer molecular surrogate subtypes to investigate the prognostic relevance and role of oxidative base lesion (8-OHdG) in the etiology of breast cancer. MATERIALS AND METHODS: 8-OHdG expression was immunohistochemicaly studied on tissue microarrays constructed from 152 patients with invasive breast cancer. Expression was correlated with other prognostic factors, as well as different breast cancer molecular surrogate subtypes such as luminal A, luminal B [human epidermal growth factor receptor 2 (HER2) negative], luminal B (HER2 positive), HER2-enriched ad triple-negative tumors. RESULTS: Triple-negative breast carcinomas (TNBCs) were more frequently 8-OHdG negative compared with non-TNBCs (P=0.036). There was no statistically significant difference between 8-OHdG expression and other breast cancer molecular subtypes.In univariate analysis, there was no significant difference between 8-OHdG expression and breast cancer-specific death, although in multivariate analysis 8-OHdG overexpression was associated with better breast cancer-specific survival (BCSS) (odds ratio=0.04, 95% confidence interval, 0.002-0.62). In Cox regression analysis, patients with moderate and strong 8-OHdG expression had 0.9 times smaller breast cancer death hazard ratio than patients with negative 8-OHdG expression. CONCLUSIONS: Oxidative stress may have less impact in the pathogenesis of TNBCs compared with other surrogate breast cancer molecular subtypes. 8-OHdG may be a promising biomarker in the prediction of prognosis for breast cancer patients.

Prognostic value of ki-67 in breast carcinoma: tissue microarray method versus whole section analysis- potentials and pitfalls.

In our study we have compared the prognostic value of two distinct methods of immunohistochemical Ki-67 determination, tissue microarray (TMA) and classical whole section analysis. "Cut-off" values were used according to the 2009 St. Gallen Consensus. Tissue specimens were obtained from a consecutive retrospective series of 215 female patients with primary invasive tumours. Two hundred and thirteen patients were included in the study. Data on Ki-67 was collected by both tissue microarray (TMA) and whole section analysis. Follow up data on overall (OS) and disease-free survival (DFS) were collected. Median follow-up was 95 months (range from 7.8 through 107 months). Mutual correlation of two Ki-67 determination methods was non-significant (Person's r = 0.13417; p = 0.0528). There was statistically significant association of whole section Ki-67 expression with histological and nuclear grade, progesterone receptor and HER2/neu status. The expression of Ki-67 protein in TMAs correlated only with histological and nuclear grade, but not with other traditional clinicopathological factors. Statistically significant differences in DFS (p = 0.0156) and OS (p = 0.0028) were confirmed between subgroups with low and high whole section Ki-67 expression. When subgroups with high and intermediate expression were compared, significant difference was found in DFS (p = 0.0272), but not in OS (p = 0.0624). On the other hand, there was no statistically significant difference either in DFS, or in OS, according to the expression of Ki-67 in TMAs (p = 0.6529; p = 0.7883; p = 0.7966 for DFS, and p = 0.8917; p = 0.6448; p = 0.4323 for OS, respectively). In our study, classical whole section was superior to TMA analysis in terms of prognosis and clinicopathological correlation. Our results indicate that the method used may have impact on prognostic significance of Ki-67. Further studies are needed, covering a greater number of patients and including a precisely defined stage and treatment patient cohorts, in order to solve controversies in Ki-67 assessment methodology.

The value of immunohistochemical determination of topoisomerase IIα and Ki67 as markers of cell proliferation and malignant transformation in colonic mucosa.

BACKGROUND: Considering the role of topoisomerase IIα in cell division, and its increased expression in the G2/M phase of cell cycle, immunohistochemical staining for topoisomerase IIα might be used as a marker for cell proliferation or malignant transformation and compared with traditional proliferation marker (Ki67). MATERIALS AND METHODS: In this study, 215 consecutive patients, who had undergone colonoscopy with biopsy, were included. The biopsy samples were fixed in formaldehyde, embedded in paraffin, cut into 3- to 4-µm sections, stained with hematoxylin-eosin and analyzed under a light microscope for pathologic diagnosis. For determination of topoisomerase IIα and Ki67 expression, paraffin blocks were cut into 4- to 5-µm sections and immunohistochemically stained with antitopoisomerase IIα antibody and Ki67. The dysplasia level was determined in adenomas and inflammatory bowel disease (IBD) samples using standard histologic criteria, whereas the grade of differentiation was determined for all colonic carcinomas. RESULTS: In 27 patients (12.6%) normal colonic mucosa was found, IBD was diagnosed in 72 patients (33.5%), 68 (31.6%) patients had adenomas, and 48 (22.3%) were diagnosed as colon carcinoma. In all samples with epithelial dysplasia, higher topoisomerase IIα expression was related to a higher level of dysplasia grade (P<0.0001). High topoisomerase IIα expression was found in all samples of colorectal carcinomas (low and high grade). There was strong correlation between topoisomerase IIα and Ki67 expression in all levels of epithelial dysplasia as well as tumor grades. CONCLUSIONS: The imunohistochemical staining for topoisomerase IIα expression might be useful as additional marker for mucosal dysplasia degree determination, as well as in differentiation of reactive/repair changes from dysplasia in IBD patients, but these findings need further evaluation.

Prognostic value of proliferation markers expression in breast cancer.

In this study, immunohistochemical expression of five proliferation markers: Ki-67, aurora-A kinase, survivin, B-Myb and cyclin B1, was analyzed. Consecutive 215 tumor samples from breast cancer patients operated from 2002 to 2003 were analyzed using the TMA ("tissue microarray") method. The median follow-up was 95 months (from 7.8 to 107 months). Statistically significant correlations between expression levels in five proliferation markers, and correlations between some of the proliferation markers and traditional prognostic factors were found. Statistically significant prognostic influence of aurora-A kinase, survivin and B-Myb expression levels on overall and disease-free survival was found, and cyclin B1 expression level on disease-free survival. A multivariate analysis confirmed survivin and B-Myb expression as independent prognostic factors of overall (p = 0.0195; p = 0.0004) and disease-free survival (p = 0.0107 and p = 0.0205) in breast cancer patients.

Biocompatibility and adhesion formation of different endoloop ligatures in securing the base of the appendix.

BACKGROUND AND OBJECTIVES: The common technique used in securing the base of the appendix is Endoloop ligature (Ethicon, Somerville, NJ, USA). Vicryl (polyglactin 910) (Ethicon) and polydioxanone (PDS) (Ethicon) Endoloop ligatures can be used. There are potential benefits of the use of PDS Plus (Ethicon) Endoloop ligature. However, the use of different materials may vary in terms of inflammation, foreign-body reaction, rate of infection in the surgical area, or rate of adhesion formation. An ideal suture would induce minimal inflammatory response and adhesion formation. METHODS: Ninety rats were randomized into 3 groups: group I, in which appendectomy was performed with Vicryl ligature; group II, in which appendectomy was performed with PDS ligature; and group III, in which appendectomy was performed with PDS Plus ligature. The animals were killed on the seventh, 28th, and 60th days after surgery. The secured stump was used for histopathologic and immunohistochemistry analysis, as well as evaluation of the formation of adhesions. RESULTS: Mild and moderate inflammation was more frequent in the PDS and PDS Plus groups than in the Vicryl group on the seventh postoperative day. There were no significant differences in the degree of inflammation on the 28th and 60th postoperative days. The lowest degree of postoperative adhesions was observed in the PDS group. CONCLUSION: Milder postoperative inflammatory changes and a lower degree of postoperative adhesions were seen in the PDS ligature group, suggesting that this could be the standard Endoloop used to secure the base of the appendix.

[Breast cancer metastases to the stomach and colon: two case reports]. / Metastaze karcinoma dojke u zeludac i debelo crijevo: prikaz dviju bolesnica.

Summary. Breast cancer has a high potential for metastasis, usually to the lungs, bones, liver and lymph nodes. Metastases in the holow organs of the digestive system are rare and mainly affectes the stomach and colon. They are characterized by very different clinical and radiological manifestations. We have warned that the initial unrecognized breast cancer can appear as a primary tumor of the stomach and colon, and onlya histopathological analysis reveales that it is a metastatic breast cancer. Metastases to the stomach or intestine involve deep layer of the mucosa and pathohistological findings of standard biopsy sample can be falsely negative, despite positive imaging technique (abdominal ultrasound and MSCT, endoscopic ultrasound) that indicate the tumor process. That's,why we emphasize the importance of endoscopic mucosal resection in the detection of malignant process of deeper layers of the gastric mucosa and deep intestinal mucosal biopsies with postoperative analysis of its walls.

Extramedullary plasmacytoma imitating neoplasm of the gallbladder fossa after cholecystectomy.

Extramedullary plasmacytomas are plasma cell tumors that arise outside of the bone marrow. They account for approximately 3% of plasma cell neoplasms and are most frequently located in the head and neck region. Five months after undergoing cholecystectomy, a 69-year-old patient presented with the pain under the right costal margin and a 12 kg weight loss. Computed tomography of the abdomen demonstrated irregular, vascular mass in the gallbladder fossa that dents towards the duodenum and the pylorus and lowers caudally to the hepatic flexure. His laboratory tests indicated normocytic anemia and showed elevated sedimentation rate. During operative procedure, a tumorous mass in the gallbladder fossa was found, inseparable of the peritoneum of the hepatoduodenal ligament and the IVb liver segment. Histopathological examination and immunohistochemical staining determined the diagnosis of the plasmacytoma. Total resection of the tumor was achieved and after 24-month follow-up patient showed no signs of local recurrence or dissemination of the disease.

Prognostic value of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and aminopeptidase N/CD13 in breast cancer patients.

The aim of this study was to analyse the expression of matrix metalloproteinase-2(MMP-2), matrix metalloproteinase-9 (MMP-9) and aminopeptidase APN/CD13 in breast carcinoma samples, and their possible prognostic value in breast cancer patients. The expression of MMP-2, MMP-9 and APN/CD13 in tumor cells was analysed in 138 breast carcinomas by immunohistochemical staining of tumor cells using the semiquantitative method for the detection of cytoplasmic and membrane reaction in tumor cells as well as stromal cells positivity. MMP-2 was positive in tumor cells of 52.9% patients and in stromal cells of 74.6% patients, while MMP-9 positive tumor and stromal cells were found in 84.8 and 63.8% patients, respectively. Tumor cell APN/CD13 expression was found in 36.2% patients. Stromal cell MMP-2 expression correlated significantly with tumor size and neoangiogenesis. A positive correlation was also observed between tumor cell MMP-9 expression and hormone receptor status. Stromal cell coexpression of MMP-2/MMP-9 correlated significantly with tumor size. APN/CD13 expression in tumor cells significantly correlated with tumor type and neoangiogenesis. Overall survival was significantly shorter in patients with MMP-2, MMP-2/MMP-9 positive tumor cells, and tended to be shorter in patients with APN/CD13 positive tumor cells. Coexpression of MMP-2/MMP-9 in tumor cells was an independent risk factor for patient survival (OD = 13.9). Our results suggest that MMP-2, MMP-9, APN/CD13 expression and MMP-2/MMP-9 coexpression in combination with other standard prognostic factors can serve as a poor prognostic factor in the evaluation of breast cancer prognosis.