Antonios Keramopoulos

Study of phospho- β -catenin subcellular distribution in invasive breast carcinomas in relation to their phenotype and the clinical outcome

β-Catenin has a crucial role in cell–cell adhesion as well as a signaling role as a member of the Wnt pathway. The aim of this study was to examine the clinicopathological and prognostic value of phosphorylated β-catenin, as well as its relation to the tumors’ phenotype, in breast cancer. Immunohistochemistry was applied on 141 paraffin-embedded breast tissue specimens for the detection of phospho-β-catenin, ER, PR, c-erbB-2, p53, Ki-67, bcl-2, uPAR and TIMP-1. For each case, a phospho-β-catenin index was determined by image analysis. Phospho-β-catenin staining was detected in the cytoplasm and the nucleus of the malignant cells. Cytoplasmic phospho-β-catenin was statistically higher in carcinomas of smaller tumor size (P=0.030), lower stage (P=0.026), decreased Ki-67 and high c-erbB-2 immunoreactivity (P=0.052 and P=0.037, respectively). Nuclear phospho-β-catenin showed a parallel correlation with ER and ERβ (P=0.022 and P=0.043, respectively), bcl-2 (P=0.042), uPAR in cancer cells (P=0.041) and TIMP-1, although the correlation was borderline (P=0.066). Cytoplasmic phospho-β-catenin was found to be independently correlated with prolonged disease-free and overall survival (P=0.046 and P=0.002, respectively), whereas nuclear localization was correlated with a shortened overall survival (P=0.046). In conclusion, phospho-β-catenin may have a different involvement in invasive breast carcinomas, according to its subcellular distribution. Nuclear localization seems to be related to an aggressive tumor phenotype, negatively affecting patients’ overall survival, whereas cytoplasmic localization is associated with a favorable tumor phenotype and a longer disease-free and overall survival.

PPARgamma expression in breast cancer: clinical value and correlation with ERbeta.

Aims : To examine the expression of peroxisome proliferator‐activated receptor γ (PPARγ) in invasive breast carcinoma in relation to known clinicopathological features, ERβ, and relapse‐free and overall patient survival. PPARγ is a ligand‐activated transcriptional factor that regulates the transcription of various target genes and has been implicated in human breast cancer.

Pregnancy and Offspring after the Appearance of Breast Cancer

The objective of this study was to investigate the influence of pregnancy in breast cancer prognosis of women under the age of 35 years. Two hundred and forty-three women with breast cancer, from thre

Overexpression of Cyclooxygenase-2 Is Associated with a Favorable Prognostic Phenotype in Breast Carcinoma

Objectives: Cyclooxygenase (COX) is the rate-limiting enzyme that catalyzes the conversion of arachidonic acid to prostaglandins. The purpose of the present study was to investigate the involvement of COX-2 protein in breast cancer biological behavior through its correlation with the well-known clinicopathological parameters and the expression of p53, c-erbB-2, topoisomerase IIα (topoIIα) and peroxisome proliferator-activated receptor (PPARγ) proteins, as well as its effect on patients’ survival. Methods: We performed immunohistochemistry to detect COX-2, estrogen receptor (ER), progesterone receptor (PR), p53, c-erbB-2, topoIIα and PPARγ proteins in 175 cases of invasive breast carcinomas. The results were elaborated by statistic analysis. Results: Cytoplasmic expression of COX-2 was detected in 66.9% of breast carcinoma samples and was inversely correlated with both nuclear and histological grade (p < 0.0001 and p = 0.039, respectively), whereas its association with PR was found to be positive (p = 0.016). COX-2 expression was inversely correlated with topoIIα and p53 (p = 0.033 and p = 0.002, respectively), whereas its association with PPARγ was parallel (p < 0.0001). In addition, c-erbB-2 of tumor cells was inversely correlated with COX-2 in stromal cells of the tumor (p = 0.011). Neither univariate nor multivariate analysis demonstrated any association between COX-2 expression and patient overall or disease-free survival. Conclusions: The current data suggest that increased expression of COX-2 may be related to breast carcinomas with less aggressive phenotype. This suggestion is further supported by the positive correlation between COX-2 and PPARγ, since the latter is considered to be indicative of a less malignant phenotype of tumor cells.

Expression of tissue inhibitor of matrix metalloproteinases (TIMP)-3 protein in invasive breast carcinoma: Relation to tumor phenotype and clinical outcome

IntroductionOur aim was to study the expression pattern of tissue inhibitor of metalloproteinases (TIMP)-3 protein in invasive breast carcinoma, and its clinicopathological and prognostic value as well as its relation to markers indicative of the tumor phenotype.MethodsImmunohistochemistry was performed on paraffin-embedded tissue specimens from 173 invasive breast carcinomas to detect the proteins TIMP-3, estrogen receptor (ER), progesterone receptor, p53, c-erbB-2, topoisomerase IIα and Bcl-2.ResultsTIMP-3 protein was immunodetected in the cytoplasm of the malignant cells and the peritumoral stroma, as well as in in situ carcinoma and normal epithelium. Reduced expression of TIMP-3 protein within cancer cells was correlated with carcinomas of high nuclear and histological grade (p = 0.032 and p = 0.015, respectively), and low ER expression (p = 0.053). Moreover, TIMP-3 immunopositivity was inversely correlated with the expression of p53 and topoIIα proteins (p = 0.002 and p = 0.008, respectively), whereas it was positively associated with Bcl-2 expression (p = 0.020). Reduced expression of TIMP-3 protein within cancer cells was found to have an unfavorable impact on disease-free survival (p = 0.052) in the entirety of the patient population, as well as in both subgroups of lymph-node-positive and mutant-p53-negative patients (p = 0.007 and p = 0.037, respectively). Stromal localization of TIMP-3 protein was found to have no clinicopathological or prognostic value.ConclusionThis is the first immunohistochemical study to show that TIMP-3 protein within cancer cells is associated with tumor phenotype. Reduced expression of TIMP-3 protein within cancer cells was found to correlate with an aggressive tumor phenotype, negatively affecting the disease-free survival of both subgroups of lymph node-positive and mutant-p53-negative patients.

The multifunctional role of the immunohistochemical expression of MMP-7 in invasive breast cancer†

The secretion of matrix metalloproteinases (MMPs) is crucial in the metastasis of cancer cells, since MMPs are responsible for the degradation of extracellular matrix (ECM). Among them, matrix metalloproteinase‐7 (MMP‐7) or matrilysin 1 is a stromelysin which degrades type‐IV collagen, fibronectin and laminin. Immunohistochemistry was performed to detect MMP‐7 protein in infiltrative breast carcinomas. MMP‐7 was studied along with clinicopathological parameters, disease‐free and overall survival, and p53, c‐erbB‐2, topoIIa, MMP‐2, uPAR and β‐catenin. MMP‐7 immunoreactivity was detected in the cytoplasm of cancer cells in 54.2% (96/177) and tumor stromal cells in 47.5% (84/177), as well as in normal epithelium adjacent to malignant epithelium. MMP‐7 reactivity in cancer cells displayed an inverse association with nuclear grade (p=0.049) and topoIIa (p=0.03). A parallel association was observed between the expression of MMP‐7 in both malignant and stromal cells with uPAR in cancer cells (p=0.033 and p=0.027, respectively). MMP‐7 of tumor stromal cells depicted a parallel correlation with MMP‐2 of the same cell type (p=0.044), while abnormal β‐catenin expression was inversely associated with MMP‐7 of cancer cells (p=0.047). Our results show the multifunctional role of MMP‐7 in the mammary gland, since it seems to be associated with a less aggressive phenotype, while, at the same time, being involved in invasion, through its collaboration with indicators of invasion.

Effect of estrogen receptor modulator tamoxifen on blood pressure, plasma renin activity, and renal sodium excretion.

BACKGROUND Adjuvant treatment with the estrogen receptor modulator tamoxifen is a well established long-term therapy in breast cancer. This study investigated the effect of tamoxifen on blood pressure (BP) and on factors by which it might be influenced. METHODS Normotensive postmenopausal women on > 12 months adjuvant tamoxifen therapy were randomized to withdraw or continue tamoxifen for 6 weeks and then to crossover to the alternative regimen for a second 6-week period. Measurements of clinic and ambulatory BP, plasma renin activity (PRA), and fractional sodium excretion (FE(Na)) were performed at baseline and at the end of each study period. RESULTS Twenty-three women completed the study (mean age 60.6 +/- 8.3 years). There was no effect of tamoxifen on clinic BP (mean difference between withdrawal and continuation for systolic BP, 0.4 +/- 8.4 mm Hg, 95% confidence interval [CI] -4.0 to 3.2, and diastolic 0.6 +/- 4.7, 95%CI -1.4 to 2.7) or 24-hour ambulatory BP (systolic 0.7 +/- 7.4 mmHg, 95%CI -2.6 to 3.9; diastolic BP, 1.9 +/- 5.5, 95% CI -0.5 to 4.2). Furthermore, no effect of tamoxifen on PRA (mean difference between withdrawal and continuation 0.03 +/- 0.5 ng/mL/h, 95% CI -0.3 to 0.2) or FENa (0.05 +/- 0.5, 95% CI -0.2 to 0.2) was detected. CONCLUSIONS Tamoxifen seems to have no effect on BP, PRA, or FE(Na) in normotensive postmenopausal women.

Sequential Administration of Doxorubicin and Paclitaxel Followed by Cyclophosphamide, Methotrexate and 5-Fluorouracil Combination (CMF) in Women with Metastatic Breast Cancer

Although the combination of paclitaxel with doxorubicin has yielded high response rates in metastatic breast cancer, severe cardiotoxic events have been reported in several patients. The rationale for our study was to evaluate the activity of paclitaxel/doxorubicin combination in patients with this disease but to avoid excessive cardiotoxicity. Therefore, we administered 4 cycles of doxorubicin/paclitaxel followed by 6 cycles of standard cyclophosphamide, methotrexate and 5-fluorouracil (CMF) regimen. Study medication consisted of doxorubicin 60 mg/m2 as a 15-min intravenous infusion followed by paclitaxel 175 mg/m2 as a 3-hour infusion. CMF regimen consisted of cyclophosphamide 600 mg/m2 as 1-hour intravenous infusion followed by methotrexate 40 mg/m2 and 5-fluorouracil 600 mg/m2 bolus injection. The main toxicity of doxorubicin/paclitaxel treatment phase was neutropenia (WHO grade 3/4, 58%), but we observed only one cardiac adverse event. Toxicities of the CMF treatment phase were not significant. Of 24 patients evaluable for response, 2 (8%) had complete responses and 11 (46%) achieved partial response. Ten additional patients (42%) had stable disease. The median time to progression was 12 months and the median overall survival was 18.5 months. The sequential administration of doxorubicin and paclitaxel followed by CMF appeared active and well tolerated in patients with metastatic breast cancer.

Pregnancy and Offspring After the Appearance of Breast Cancer

The objective of this study was to investigate the influence of pregnancy in breast cancer prognosis of women under the age of 35 years. Two hundred and forty-three women with breast cancer, from three oncology departments in Athens, were investigated. Twenty-one got pregnant (7.91%) 7-100 months after breast cancer diagnosis and in a median time period of 31 months. All women had mastectomy apart from 2 who had only lumpectomy as surgical procedure. Thirteen of 21 were treated with radiotherapy and 17 of 21 had also adjuvant chemotherapy mainly with CMF for 6 cycles. Sixteen children from 14 mothers were born and the rest of the patients underwent an abortion between the 2nd and 5th month of pregnancy. All children were healthy and grew up normally up to the age of 12-142 months (end of the study) and their median age of 51 months. Only 2 patients had stage III disease at diagnosis while the remaining 19 had stage I-IIb. Three cancer recurrences were observed (14.3%) after 7-84 months. One patient had a second primary-ovarian cancer 60 months after mastectomy. Recurrence rate and survival compared with those of nonpregnant women of the same age and the stages of disease were not different. To conclude: the present study indicates that healthy offsprings can be delivered from breast cancer patients, and pregnancy does not seem to play any role in tumor recurrence.