Breast Neoplasms: Simopoulos C

Giatromanolaki A, Koukourakis MI, Simopoulos C, Sivridis E. · Department of Pathology, Democritus University of Thrace Medical School, Alexandroupolis 68100, Greece. · Cancer Biol Ther. · Pubmed #17224643 No free full text.

Abstract: The expression of c-erbB-2 and HIF1alpha proteins is linked with an aggressive tumor phenotype and poor survival in breast cancer. In the present study we investigated whether this ominous effect is a result of c-erbB-2/HIF1alpha expressing clone appearance within the primary tumor, by examining the c-erbB-2/HIF1alpha expression status in the primary tumor, node metastasis and cancer cells invading into the lymphovascular spaces. The metastasizing cancer cell clones, whether migrating to the lymph nodes or entering into the systemic circulation maintained the original c-erbB-2/HIF1alphaphenotype of the primary tumor. Migrating c-erbB-2/HIF1alpha negative tumors do so through activation of alternative biologic pathways and not through positive clone appearance. These results strongly support the concept that targeted therapies against c-erbB-2 or against HIF1alpha can be guided with precision by the primary tumor c-erbB-2/HIF1alpha status without demanding the detection of the metastatic tumor phenotype.

Koukourakis MI, Simopoulos C, Polychronidis A, Perente S, Botaitis S, Giatromanolaki A, Sivridis E. · Departments of Radiotherapy-Oncology, Surgery and Pathology, Democritus University of Thrace, Medical School, Alexandroupolis 68100, Greece. · Anticancer Res. · Pubmed #12820439 No free full text.

Abstract: In this study we describe and discuss the dichotomous effects of docetaxel trastuzumab (Herceptin)/docetaxel therapy on the angiogenic molecular profile in two patients with her-2 + chemo-resistant recurrent breast carcinoma. In the first case, an intensification of angiogenesis occurred following therapy, accompanied by an impressive increase of the cancer cell proliferation index. This tumor did not express HIF1 alpha and shared a HIF-independent VEGF overexpression, which remained unaffected by therapy. An intensified formation of thymidine phosphorylase (TP)-rich stroma, presumably a response to docetaxel, shows a TP-dependent angiogenic response. In the second patient, VEGF and HIF1 alpha were down-regulated in post-treatment biopsies and this was accompanied by a sharp reduction of the vascular density and of the cancer cell proliferation rate. In both cases, c-erbB-2 expression was abrogated by Herceptin. Taking into account that Herceptin down-regulates VEGF through reduction of HIF1 alpha synthesis, this clinical study provides evidence that an anti-angiogenic effect from Herceptin/Docetaxel therapy is expected only in tumors with HIF1 alpha-dependent VEGF overexpression. In contrast, HIF1 alpha-independent VEGF angiogenic activity cannot be abrogated by Herceptin. Docetaxel mediated up-regulation of TP in the tumoral stroma may, on the contrary, result in angiogenesis intersification and rapid tumor relapse. Such an effect should be of clinical importance since Herceptin/Docetaxel-based regimens are currently evaluated for the adjuvant therapy of her-2 + breast cancer patients. Studying the Herceptin-induced phenotypic changes of tumors could lead to the identification of specific molecular profiles that bring about diverging angiogenic responses. Adjustment of the chemotherapy regimen accordingly would prove of clinical importance.

Limberis V, Romanidis C, Galazios G, Koutsougeras G, Papadopoulos N, Lambropoulou M, Simopoulos C. · Department of Obstetrics & Gynecology, Democritus University of Thrace, Greece. · Eur J Gynaecol Oncol. · Pubmed #19317265 No free full text.

Abstract: BACKGROUND: Frozen section biopsy has been widely used for intraoperative diagnosis and evaluation of sentinel lymph nodes, so a decision can be made regarding whether to perform axillary clearance during primary surgery. This study aims to discuss the reliability of a simpler and faster method - touch imprint cytology - in the interpretation of metastasis from breast cancer. METHODS: A retrospective review of 41 sentinel lymph node biopsies from patients with breast cancer were examined by intraoperative imprint cytology using rapid Diff-Quick staining. Paraffin-embedded permanent sections were examined using hematoxylin and eosin stained sections from the sentinel lymph nodes in collaboration with the employment of an anti-cytokeratin antibody. RESULTS: Sixteen of all sentinel nodes harbored metastases in the paraffin sections, of which all 16 were identified by imprint cytology (sensitivity 93%). CONCLUSION: Touch imprint cytology is a fast and reliable alternative for intraoperative evaluation of sentinel lymph nodes in breast cancer patients.

Koukourakis MI, Manavis J, Simopoulos C, Liberis V, Giatromanolaki A, Sivridis E. · Department of Radiotherapy/Oncology, Democritus University of Thrace, Alexandroupolis, Greece. · Am J Clin Oncol. · Pubmed #16199990 No free full text.

Abstract: OBJECTIVES: Trastuzumab, an antic-erbB-2 monoclonal antibody, has become a standard component of chemotherapy for c-erbB-2-positive advanced breast carcinoma. Despite the experimental evidence of its radiosensitizing properties, trastuzumab has never been used in combination with radiotherapy for the treatment of patients with locally advanced disease. PATIENTS AND METHODS: Twenty-two patients with c-erbB-2-positive locally advanced chemoresistant (7 patients) or with high-risk breast cancer (15 patients) were recruited in a treatment protocol combining hypofractionated/accelerated radiotherapy (hypoARC) supported with high-dose amifostine (1000 mg subcutaneous), concurrently with trastuzumab (4 mg/kg every 2 weeks). Thirteen of these patients (including all 7 inoperable cases) received concurrently chemotherapy with liposomal doxorubicin and docetaxel (25 mg/m2 and 40 mg/m2 every 2 weeks, respectively). RESULTS: Administration of trastuzumab together with highly accelerated amifostine-supported radiotherapy was feasible without an increase in early and late radiation toxicity. This was obtained despite the concurrent administration of aggressive chemotherapy. Complete responses were noted in 5 of 7 patients with locally, often far advanced, chemoresistant disease. None of the complete responders or the 15 high-risk breast cancer patients relapsed within the 3- to 26-month follow-up period. CONCLUSION: Inclusion of trastuzumab in the radiochemotherapy protocols for breast cancer does not increase radiation or systemic toxicity. The concurrent administration of aggressive radiotherapy with docetaxel and liposomal doxorubicin is feasible when supported with amifostine. The value of such regimens in the treatment of locally advanced or high risk c-erbB-2 positive breast cancer patients deserves further evaluation.

Tamiolakis D, Venizelos I, Nikolaidou S, Prassopoulos P, Alexiadis G, Simopoulos C, Papadopoulos N. · Department of Cytology, General Hospital of Chania, Crete, Greece. · Onkologie. · Pubmed #15585977 No free full text.

Abstract: BACKGROUND: Rhabdomyosarcoma accounts for approximately 4% of all childhood malignancies. Breast metastases from rhabdomyosarcoma are uncommon with an incidence of 6%. CASE REPORT: We present a patient who developed bilateral mammary metastases from rhabdomyosarcoma arising in the right lower extremity. An 11-year-old female with a 20-month history of rhabdomyosarcoma was referred to our department because of bilateral breast enlargement. A needle core biopsy was performed and touch imprint slides were obtained. Cytology determined the masses to be metastases of rhabdomyosarcoma. MyoD1 immunostain and RT-PCR analysis confirmed the diagnosis. CONCLUSIONS: Cytomorphology with ancillary methods is essential in the diagnosis of metastatic breast deposits in order to avoid unnecessary mastectomy and to employ systemic treatment.

Giatromanolaki A, Koukourakis MI, Simopoulos C, Polychronidis A, Gatter KC, Harris AL, Sivridis E. · Department of Pathology, Medical School, Democritus University of Thrace, Alexandroupolis, Greece. · Clin Cancer Res. · Pubmed #15585632 links to  free full text

Abstract: c-erbB-2-positive breast carcinomas are highly aggressive tumors. In vitro data on breast cell lines showed that c-erbB-2 enhanced translational efficiency of hypoxia inducible factor-1alpha (HIF1alpha) production (Laughner et al., Mol Cell Biol 2001;21:3995-4005). We investigated the clinical correlate of this observation to assess whether c-erbB-2 expression was related to HIF1alpha expression, angiogenesis, and prognosis. A series of 180 breast carcinomas of known c-erbB-2 status (90 c-erbB-2-positive and 90 c-erbB-2-negative carcinomas) were stained immunohistochemically for HIF1alpha and CD31 endothelial cell antigen. c-erbB-2 positivity was clearly related to HIF1alpha protein expression and high angiogenesis. However, prognosis was decreased only in cases with simultaneous c-erbB-2 and HIF1alpha expression. If activation of c-erbB-2 in humans results in overexpression of HIF1alpha independently of conditions of hypoxia, as occur in experimental studies, this interaction may represent a main pathway conferring clinical aggressiveness to c-erbB-2-positive breast tumors.

Koukourakis MI, Giatromanolaki A, Chong W, Simopoulos C, Polychronidis A, Sivridis E, Harris AL. · Departments of Radiotherapy/Oncology, Democritus University of Thrace, 68100, Alexandroupolis, Greece. · Cancer Chemother Pharmacol. · Pubmed #14574457 No free full text.

Abstract: PURPOSE: The cytoprotective mechanism of amifostine (WR-2721) implies free radical scavenging and DNA repair activities. We investigated additional cytoprotective pathways involving intracellular hypoxia and the activation of the hypoxia-inducible factor (HIF) pathway, a key transcription factor regulating glycolysis, angiogenesis and apoptosis, which is also linked with radioresistance. MATERIALS AND METHODS: The glucose and oxygen levels in the peripheral blood of patients receiving 1000 mg amifostine were determined at various time-points in order to investigate the metabolic changes induced by amifostine. MDA468 breast tumor cell lines were incubated with a high amifostine concentration (10 m M) to overcome the natural resistance of cancer cells to influx of the non-hydrolyzed WR-2721, and the HIF1 alpha protein levels were determined by Western blot analysis. In vivo experiments with Wistar rats were performed in order to assess immunohistochemically changes in the intracellular accumulation of HIF1 alpha induced by amifostine (200 mg/kg). RESULTS: By 30 min following amifostine administration, the hemoglobin oxygen saturation and pO(2) levels had increased in the peripheral blood while glucose levels had reduced, providing evidence that normal tissue metabolism switches to glycolytic pathways. Incubation of cell lines with amifostine resulted in HIF1 alpha induction. In Wistar rats administration of amifostine resulted in increased HIF1 alpha accumulation in normal tissues. CONCLUSIONS: Since it is doubtful whether dephosphorylation of amifostine to the active metabolite WR-1065 occurs within tumoral tissues (an acidic environment that lacks vascular alkaline phosphatase activity), intracellular hypoxia and upregulation of HIF1 alpha represents an additional, normal tissue-specific, amifostine cytoprotective pathway.

Giatromanolaki A, Sivridis E, Simopoulos C, Polychronidis A, Gatter KC, Harris AL, Koukourakis MI. · Tumour and Angiogenesis Research Group, Department of Radiotherapy/Oncology, Democritus University of Thrace, Alexandroupolis, Greece. · Clin Exp Metastasis. · Pubmed #12553372 No free full text.

Abstract: Recent reports provide evidence that some growth factors behave as inhibitors of the apoptosis of the endothelial cells, bringing forward the concept of vascular survival as a post-angiogenesis process. At least two different vasculature development processes occur within a tumor: the angiogenic (formation of new vessels) and the vascular survival pathway, which is devoted to the preservation of the newly-formed vessels in layers that lose contact with the adjacent normal tissue. We developed a method to assess these processes in tissue samples. We noted that differences among tumors may exist not only in the tumor angiogenic activity (TAA) but also in the vascular survival ability (VSA). One third of the highly angiogenic breast cancer cases examined had a poor ability to maintain high vessel density in inner tumor areas. Both parameters are independently related to prognosis, while VSA was directly related to tumor dimensions and node involvement. Patients with high TAA and VSA had a particularly poor prognosis. It is suggested that although cancer angiogenic activity is important for the local invasion and dissemination into vessels and lymphatics, the VSA may be important for the effective formation of viable tumor foci in lymph nodes or distant organs. Recognition and quantification of the vascular survival ability in human tumors may significantly improve the prognostic value of the assessment of tumor vasculature, and may help to stratify patients for clinical trials with novel anti-angiogenic or angiotoxic drugs. Elucidation of the pathways may provide additional targets for antiangiogenic therapy.

Tamiolakis D, Papadopoulos N, Cheva A, Lambropoulou M, Kotini A, Jivannakis T, Simopoulos C. · Department of Cytology, Regional Hospital of Alexandroupolis, Greece. · Eur J Gynaecol Oncol. · Pubmed #12440829 No free full text.

Abstract: Myoepithelial cells are normally located between the epithelial cells and the basal lamina of secretory elements of exocrine glands. Their role in the histogenesis of breast tumours has been studied extensively, and a definite differentiation towards myoepithelial cells has been demonstrated in adenoid cystic carcinoma, adenomyoepithelioma, low-grade adenosquamous (syringomatous) carcinoma, pure malignant myoepithelioma and poorly differentiated myoepithelial-rich breast carcinoma. All these tumours are of low malignancy, with the exception of malignant myoepithelioma and poorly differentiated myoepithelial-rich carcinoma. We examined the possibility that invasive ductal carcinoma of the breast might show differentiation towards both epithelial and myoepithelial cells because there is no reason to assume that one type of differentiation necessarily excludes the other. We performed the avidin-biotin immunohistochemical analysis of 20 cases of infiltrating ductal carcinomas (IDCs) with diffuse fibrosis, 20 cases of IDCs without fibrosis and five cases of metaplastic carcinomas, to detect myoepithelial differentiation of the tumour cells. Myoepithelial differentiation was determined by the expression of alpha-smooth muscle actin (alpha-SMA). We concluded that IDCs with diffuse fibrosis are associated with a myoepithelial immunophenotype of carcinoma cells.

Tamiolakis D, Simopoulos C, Cheva A, Lambropoulou M, Kotini A, Jivannakis T, Papadopoulos N. · Department of Cytology, Regional Hospital of Alexandroupolis, Democritus University of Thrace, Greece. · Eur J Gynaecol Oncol. · Pubmed #12440819 No free full text.

Abstract: Medullary carcinoma (MC) of the breast is considered to carry a more favorable prognosis than other subtypes of infiltrating ductal carcinoma. This is a biological paradox because its clinical behavior contrasts with its anaplastic morphology. MC is characterized by a dense lymphocytic infiltrate. In this study, we determined the immunological profile of tumor infiltrating lymphocytes (TILs) in MC by CD20 (L26), CD8, and CD45RO (UCHL 1) immunostaining on paraffin-embedded sections. We examined 14 cases of typical MC (TMC), 15 cases of atypical MC (AMC) classified according to Ridolfi criteria (1977) and 19 cases of poorly differentiated infiltrating ductal carcinoma (PDC-NOS). TILs were quantified separately into cells infiltrating tumor nests (intraepithelial) and cells infiltrating tumor stroma (stromal). The number of CD8 positive and CD20 positive cells infiltrating tumor nests and tumor stroma were significantly increased in TMC and AMC as opposed to the PDC-NOS group. There was a loss in the number of CD45RO positive cells, both intraepithelial and stromal, in TMC and AMC as opposed to the PDC-NOS group. We conclude that MC tumor lymphocytic infiltrate demonstrates a mixed-T cytotoxic (CD8+) and B cell (CD20+)-immunophenotypic profile. This might in part explain the improved clinical outcome of the disease.

Tamiolakis D, Papadopoulos N, Venizelos J, Lambropoulou M, Romanidis C, Petrakis G, Limberis V, Galazios G, Koutsougeras G, Simopoulos C. · Department of Cytology, Regional Hospital of Chania, Greece. · Onkologie. · Pubmed #16974114 No free full text.

Abstract: BACKGROUND: Imprint cytology may provide a fast and accurate method for intraoperative screening of sentinel lymph nodes, so a decision can be made regarding whether to perform axillary clearance during primary surgery. If the findings are negative, in many cases axillary dissection can be omitted. Patients and METHODS: 128 sentinel nodes from a cohort of 87 patients that had been identified using technetium-99m nanocolloid as a radioactive tracer and Patent blue dye were dissected for rapid Diff-Quick stained touch preparations. Intraoperative evaluation of sentinel node status by imprint cytology was correlated with histopathological results of permanent sections. Tumor-negative nodes in routine paraffin sections were further investigated with the employment of an anti-cytokeratin antibody. RESULTS: 36 of all sentinel nodes harbored metastases in the paraffin sections, of which 32 were identified by imprint cytology (sensitivity 88.8%). 3 sentinel nodes were positive by imprint cytology and negative by histopathology of the paraffin sections. Comparison of the results of the touch preparations with the final histopathology (hematoxylin-eosin and anticytokeratin antibody stains) demonstrated a sensitivity of 83.3% and a negative predictive value of 92.5%. The specificity and positive predictive value were 100% each. CONCLUSIONS: Touch imprint cytology is potentially useful for intraoperative evaluation of sentinel lymph nodes in breast cancer patients.