Jan Bręborowicz

Novel BRCA1 mutations and more frequent intron-20 alteration found among 236 women from Western Poland

Three different novel BRCA1 mutations, five independent cases of the same 12 bp insertion-duplication in intron-20 and two novel rare BRCA1 sequence variants were identified among 122 Polish women with positive, in most cases moderate family history of breast and/or ovarian cancer, 80 controls and 34 unselected breast cancer tissue specimens. All mutations and variants were germline. The 4153 delA frameshift mutation, the Tyr105Cys missense mutation and two cases of the alteration in intron-20 were found in the group of healthy women with positive family history. Two other cases of the intronic insertion were found in unselected controls. Their carriers had no family history of breast or ovarian cancer but other cancers occurred in their families. The 1782 Trp/STOP nonsense mutation and one case of the insertion in intron-20 were first found in tissue specimens of breast cancer patient and breast/ovarian cancer patient, respectively. Their carriers also had no family history of breast or ovarian cancer. The distribution of the insertion in intron-20 in analysed groups and results of RT – PCR experiments suggest a less prominent role for this variant considered earlier a splicing mutation. This study shows also, that more population-oriented research is needed, involving women with less profound or even no family history of breast and ovarian cancer, to better understand the role and significance of different BRCA1 variants and mutations.

Expression of estrogen receptor beta in the breast carcinoma of BRCA1 mutation carriers.

BackgroundBreast cancers (BC) in women carrying mutations in BRCA1 gene are more frequently estrogen receptor negative than the nonhereditary BC. Nevertheless, tamoxifen has been found to have a protective effect in preventing contralateral tumors in BRCA1 mutation carriers. The identification of the second human estrogen receptor, ERβ, raised a question of its role in hereditary breast cancer. The aim of this study was to assess the frequency of ERα, ERβ, PgR (progesterone receptor) and HER-2 expression in breast cancer patients with mutated BRCA1 gene and in the control group.MethodsThe study group consisted of 48 women with BRCA1 gene mutations confirmed by multiplex PCR assay. The patients were tested for three most common mutations of BRCA1 affecting the Polish population (5382insC, C61G, 4153delA). Immunostaining for ERα, ERβ and PgR (progesterone receptor) was performed using monoclonal antibodies against ERα, PgR (DakoCytomation), and polyclonal antibody against ERβ (Chemicon). The EnVision detection system was applied. The study population comprised a control group of 120 BC operated successively during the years 1998–99.ResultsThe results of our investigation showed that BRCA1 mutation carriers were more likely to have ERα-negative breast cancer than those in the control group. Only 14.5% of BRCA1-related cancers were ERα-positive compared with 57.5% in the control group (P < 0.0001). On the contrary, the expression of ERβ protein was observed in 42% of BRCA1-related tumors and in 55% of the control group. An interesting finding was that most hereditary cancers (75% of the whole group) were triple-negative: ERα(-)/PgR(-)/HER-2(-) but almost half of this group (44.4%) showed the expression of ERβ.ConclusionIn the case of BRCA1-associated tumors the expression of ERβ was significantly higher than the expression of ERα. This may explain the effectiveness of tamoxifen in preventing contralateral breast cancer development in BRCA1 mutation carriers.

Immunoexpression of TTF-1 and Ki-67 in a coexistent anaplastic and follicular thyroid cancer with rare long-life surviving

We report the immunohistochemical diagnosis, including TTF-1 (thyroid transcription factor 1) and Ki-67, of a rare mixed thyroid neoplasm composed of minimally invasive well differentiated follicular areas and highly aggressive undifferentiated anaplastic areas. A 75 old female presented to our clinic with a rapidly growing neck mass. Considering the dynamics of the disease and the multiple challenges presented by the patient: advanced age, tumor size, history of a longstanding goiter we decided to transfer her to the department of surgery. The intraoperative findings were an enlarged right lobe with tracheal and surrounding tissues infiltration. Total thyroidectomy, radical neck lymph nodes dissection and tracheostomy were performed. The histopathological and immunohistochemical examination revealed a coexistent anaplastic and follicular thyroid carcinoma. The proliferation index Ki-67, a cell proliferation marker, was found to be significantly higher in the anaplastic areas (30 +/- 5%) in the comparison with the follicular areas (2 +/- 1%). The evaluation of the thyroid transcription factor 1 (TTF-1) expression revealed a correlation with the tumor cells aggressiveness accordingly to the cancer areas. After a radical surgery an external adjuvant radiation was applied. The patient is alive and more than five years after diagnosis she presented an increase of the serum thyroglobulin level suggesting, probably, a recurrence of the follicular form of the cancer. According to our survey we suggest that in thyroid cancers TTF-1 and Ki-67 could provides useful information on the differentiation activities of thyroid tumor cells and may be helpful to distinguish well differentiated and undifferentiated areas in a mixed thyroid cancer.

Thyroid nodule as a first manifestation of Hodgkin lymphoma-report of two cases and literature review.

AbstractLymphomas account for less than 5% of thyroid malignant lesions. Vast majority of them are B-cell non-Hodgkin lymphomas (NHL), while Hodgkin lymphoma (HL) is extremely rare. Here we present two cases of HL, at baseline manifesting as a thyroid lesion. First patient, 29-year-old pregnant female, initially suspected for metastatic medullary thyroid cancer, was eventually diagnosed with mixed cellularity type of thyroid HL. Second patient, 22-year-old woman with suspicion of advanced thyroid cancer, was in the end diagnosed with an extra-lymphatic classical HL of the thyroid. In both cases, despite repeated fine-needle aspiration biopsy, cytological examination gave inconclusive or misleading results. On histopathological examination, thyroid tumor cells were positive for CD15 and CD30 antigen, which is typical for Reed-Sternberg cells. In the report authors also discuss difficulties in management as well as potential importance of novel methods such as FISH, PCR and other molecular techniques in diagnostics of thyroid lymphomas.Virtual slidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2896947559559648

The Determination of VEGF and MVD, among Patients with Primary Breast Cancer

The purpose of the study was to ascertain the value of assessment of vascular endothelial growth factor (VEGF) levels and microvessel density, and to search for correlations between them, in women with breast cancer. The assessment considered factors such as the stage of clinical disease advancement—according to International Union Against Cancer, the grade of histological malignancy, status of axillary lymph nodes and the size of the primary tumour. The concentration of VEGF was assessed in the plasma of 103 women with breast cancer, using an immunoenzymatic method (Quantikine test of R&D Systems). Assessment of microvessel density was performed using histopathological immunoperoxidase methods, using an anti-von Willebrand factor antibody (DAKO A/S). A statistically significant relationship was found between rising VEGF levels and microvessel density in women with breast cancer, when compared to values from a control group. A correlation was found between VEGF concentration and microvessel density (MVD) values. Statistically significant differences were found between VEGF levels of patients in stages I, II and III of clinical disease advancement. For MVD, differences were found only between stages I and III. A statistical relationship was also found between VEGF and MVD and tumour size. Similar results were found between VEGF concentrations in women with metastases to the axillary lymph nodes and cytokine levels in women with no metastases. The results of the study suggest that the degree of tumour vascularization and the concentration of VEGF may represent valuable indicators for the assessment of the angiogenic process in women with breast cancer.

Evaluation of the Sentinel Node Biopsy in Colorectal Carcinoma Including the Results of Immunohistochemical Examinations

Abstract Aim: To evaluate the method and results of sentinel node biopsy including immunohistochemical examinations in resectable colorectal cancer. Material and methods: From April 2004 to April 2005, sentinel node biopsy was carried out with the dye method in 27 patients operated on for colorectal cancer. The standard examination of sentinel nodes consisted in the evaluation of individual H&E-dyed specimens from bisection of the node. The negative sentinel nodes were examined with the use of immunohistochemistry against cytokeratins AE1/AE3. Findings: The sentinel node was identified in 25 patients (92.6%). In a routine histopathological examination it included metastases in 3 cases. The sentinel node was clean in one patient whereas other regional nodes resected “en bloc” with the tumour included metastases. The sensitivity of the method was 75%, and the number of falsely negative results was 25%. Metastases (micrometastases) in the sentinel node were found in 2 other patients (8%) in the immunohisto-chemical examination. This examination did not change the results of the analysis in the patient with positive non-sentinel nodes and with the negative sentinel node in H&E dyeing. Nevertheless, the sensitivity of the method rose to 83.3% and the number of falsely negative results dropped to 16.7%. Conclusions : 1. The sentinel node biopsy using the dye method is a safe and relatively easy technique showing a high success rate (92.6%). 2. Low sensitivity of the method, as reflected in the literature, may result from lack of extended histopathological examinations performed on the sentinel node (e.g. immunohistochemistry). 3. Further research is necessary to determine the role and importance of the sentinel node biopsy in colorectal cancer.

Poststreptococcal syndrome mimicking conjunctival lymphoma

BackgroundPoststreptococcal syndrome (PSS) can be a consequence of nonpurulent primary infection with group A streptococci (GAS). Postreptococcal uveitis is a well recognized entity with quite a few descriptions in the literature, but so far no conjunctival involvement has been reported.The aim of the study is to present a rare case of postreptococcal conjunctival lesions mimicking a lymphoma.Case presentation19-years-old Caucasian female presented with pink, nodular infiltrates in the right conjunctiva that occurred a few months after upper respiratory tract infection and tonsillectomy. Histopathological examination of collected lesion samples revealed inflammatory reaction with lymphocytes proliferation and failed to rule out a myeloma. Complementary flow-cytometry did not show monoclonal proliferation of lymphocytes B. During follow-up we observed the complete regression of conjunctival lesions after the benzyl penicillin treatment prescribed by ENT specialist due to elevated plasma ASO levels. Therefore, we suppose that those lesions must have represented a part of poststreptococcal syndrome.ConclusionsTo conclude, this is, to the best of our knowledge, the first report of conjunctival involvement in the course of PSS related to group A streptococci.

Abstract 254: Expression of the putative stem cell marker CD44 in prostate biopsies correlates with Gleason score in radical prostatectomies in patients with prostate cancer.

Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC AIM: Prostate cancer is one of the leading causes of death among male oncology patients. There is growing evidence to support the hypothesis that cancer cells in the prostate are of stem cell origin. In this study we used CD44, as one of the most promising putative stem cell markers, to evaluate the prevalence of prostate cancer cells with stem cell like features in prostate biopsies and in radical prostatectomy specimens. We tested both types of specimen for the existence of a correlation between the expression of CD44 and commonly used prostate cancer prognostic factors such as Gleason grade, pathological stage (pT) according to TNM, patient age and preoperative plasma PSA levels. M&M: Formalin fixed paraffin embedded tissue samples from fifty two prostate biopsies and matched radical prostatectomy specimens were retrieved from the archives of the Pathology Department at the City Hospital, Poznan, Poland or from a collection of cases sent for consultation in the private laboratory of one of the authors (JB). Immunohistochemistry was performed on sections from all paraffin blocks using an anti-CD44 primary antibody (Abcam) according to the manufacturers protocol. Using an OLYMPUS BX41 microscope, we counted at least 100 cancer cells from core biopsies and 500 cancer cells from radical prostatectomy specimens. Only cells with clearly visible membranous staining at 10x magnification were called positive for the expression of CD44. The percentage of cancer cells expressing CD44 was used for further analysis. The statistical analysis was based on Spearman and Person correlations performed in Microsoft Office Excel 2007. RESULTS: CD44 positive cancer cells were identified in forty three prostate biopsies (82%) and forty seven radical prostatectomy specimens (90%). There was a positive correlation between the expression of CD44 in cancer cells from prostate biopsies and in radical prostatectomy specimens. We also observed a negative correlation between CD44 expression in cancer cells and Gleason score, both in prostate biopsies and in radical prostatectomy material. When other clinical data were analysed, such as patient age, preoperative PSA serum levels or tumour stage (pT), only the first parameter was found to correlate positively with CD44 expression in prostate biopsies. CONCLUSION: To the best of our knowledge this study showed for the first time, that the level of CD44 expression in prostate biopsies correlates with that observed in matched radical prostatectomy specimens. Additionally, we confirmed the results of other authors which show that the percentage of CD44 expressing cancer cells in prostate biopsies negatively correlates with Gleason grade of tumours, both in prostate biopsies and in radical prostatectomy material. Citation Format: Konstanty Korski, Anna Malicka, Jan Breborowicz. Expression of the putative stem cell marker CD44 in prostate biopsies correlates with Gleason score in radical prostatectomies in patients with prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 254. doi:10.1158/1538-7445.AM2013-254

Prognostic factors in melanoma

Summary Background There are clinical and pathological factors associated with the clinical course of melanoma. These parameters allow us to predict survival times and establish a course of treatment. Aim The aim of the study was to assess the significance of immunohistochemical markers in the progression of melanoma, and relate these findings to the influence of clinical and histological factors on survival time. Materials/Methods In this study, archival histological material obtained from 50 melanoma patients operated on in the Great Poland Cancer Centre between 1990–1995 was analysed. Using immunohistochemistry we detected the presence of markers known to be important in the diagnosis of melanoma, including: HMB-45, PCNA, Ki-67, MMP-2, CD44 and nm23. Following this, univariate logistic regression was performed and clinical, histopathological and immunohistochemical data of statistical significance were correlated with survival time using the Cox nonparametric proportional hazard regression model. Results The results suggested that the most important prognostic factors correlating with survival time were: the presence of nm23 antigen (p = 0.0342) and the thickness of the melanoma, as measured by the Breslow method in mm (p = 0.0191). According to the univariate analysis there were correlations between patient death or survival and the histological type, Superficial Spreading Melanoma Malignum (SSMM) (p = 0.0187), regional lymphatic metastases (p = 0.0030) and positive Ki-67 results (p = 0.0282). Conclusions Taken together, our results allowed us to conclude (a) that there was a correlation between survival time and nm23 antigen expression and the thickness of melanoma, as measured by the Breslow method in mm, (b) that there were correlations between patient survival or death and the histological type of Superficial Spreading Melanoma Malignum, regional lymphatic metastases and positive Ki-67, (c) that other parameters (age, gender, anatomical location of the melanoma, the presence of ulceration, lymphatic infiltration, the existence of satellites, and positive PCNA and MMP-2) showed no significant influence on survival.

The role of oestrogen receptor β in invasive breast cancer

Summary Background The presence of estrogen receptor (ER) in breast cancer cell is a good prognostic factor. It is also a predictive factor used in hormonal therapy. Aim To evaluate the expression of oestrogen receptors α and β (ERα and ERβ) in the neoplastic tissues of patients with invasive breast cancer and to determine whether ERβ expression may be correlated with some clinical parameters and biological markers. Materials/Methods Paraffin embedded tissues from 67 patients with breast cancer were used in this study. Monoclonal antibodies against α oestrogen receptors, progesterone receptors (DakoCytomation) and polyclonal antibodies against β oestrogen receptors (CHEMICON) were used. The EnVision detection system was applied. Results Expression of ERα was demonstrated in 57% of all patients, while in patients over 50 years it was higher – 71%. Expression of ERβ was demonstrated in 48% of patients and this percentage was not dependent on the age of the patients. In tumours expressing ERβ, expression of p53 and Ki-67 was less common. In addition, these tumours were of a lower grade of malignancy. Conclusions Our results demonstrate a negative correlation between the expression of ERβ and the expression of p53 and Ki-67. The expression of ERβ may be a good prognostic indicator.