Cases Of Primary Breast Carcinoma Research Articles

The role of androgen receptors in breast cancer

BackgroundBreast cancer is a heterogenous group of diseases with steroid hormone dependant carcinogenesis. Along with Estrogen Receptor (ER), Progesterone Receptor (PR), Human Epidermal growth factor Receptor 2 (HER2), androgen receptor (AR) is now emerging to play key role in disease progression of various molecular breast cancer subtypes especially triple negative breast cancers. Aim of this study was to find out prevalence of AR positivity in all breast cancers, correlation of ER, PR, HER2 and Ki67 index with AR positivity. To find out correlation of AR positivity with histological grade. MethodsThis was a descriptive cross-sectional study done on 55 primary breast carcinoma cases. Routine Hematoxylin and Eosin stain as well as immunohistochemical ER, PR, HER2/neu, Ki67 and AR stain were done. The outcome was assessed in the form of relationship between AR and ER, PR, HER2 status and Ki-67 index and histopathological grade. ResultsAR positivity was seen in 69% of all breast cancers. AR positivity was seen in 8% of triple negative breast cancers. High Ki67 index showed 55% AR positivity. Grade II breast cancers showed 42% AR positivity. ConclusionCases with triple negative breast cancers have worst prognosis. Unlike targeted novel hormone therapy (tamoxifen) in luminal cancers and monoclonal antibody (trastuzumab) in her2 enriched cancers, AR is looked upon as a target to treat triple negative breast cancer. However, the existing controversies demand more AR related studies.

Detecting Human Epidermal Growth Factor Receptor 2 (HER2) Amplification: Proof of Concept of an Alternative Approach.

There are multiple genes that are co-amplified along with human epidermal growth factor receptor 2 (HER2) in chromosome 17. GRB7 and PGAP3 are two such genes. We hypothesize that the protein products of these genes may serve as immunohistochemistry (IHC) markers for detecting HER2 amplification in breast cancer. Tissue sections from one hundred and thirty-five primary breast carcinoma cases were subjected to immunohistochemical staining for antibodies against HER2, GRB7, and PGAP3 and graded on a scale of 1 to 3. Both membranous staining and cytoplasmic staining were assessed for GRB7 and PGAP3. For equivocal HER2 IHC positivity, fluorescent in situ hybridization was performed to get the final HER2 status. IHC staining for GRB7 and PGAP 3 was a moderate to strong predictorfor HER2 status (area under the curve (AUC) of 0.768, 0.868,0.754, and 0.790 for GRB7 membranous staining, GRB7 cytoplasmic staining, PGAP3 membranous staining, and PGAP3 cytoplasmic staining respectively). A combination of GRB7 cytoplasmic and PGAP3 membranous staining resulted in an AUC of 0.905 (95% CI0.855-0.954), while a combination of GRB7 and PGAP3 cytoplasmic staining resulted in an AUC of 0.902 (95% CI 0.851-0.953). The point estimates for the AUC of GRB7 and combined GRB7 and PGAP3 in predicting the AUC suggest a strong predictive ability of these markers to predict HER2. With further refinement in technique, cytoplasmic staining and membranous IHC staining for GRB7 and PGAP3 have potential to serve as surrogate markers for HER2 status. The strategy of using protein products of co-amplified genes of HER2 is likely to be successful in technical validation.

Ki67-PROLIFERATIVE INDEX IN BREAST CANCER AND ITS ASSOCIATION WITH MOLECULAR SUBTYPE, HISTOPATHOLOGICAL GRADE & TYPE - IN A TERTIARY CARE HOSPITAL.

Introduction: Carcinoma of breast is the most frequent malignancy in women worldwide and the second most common cause of fatality in patient with cancer. Prognostic factors are essential in breast cancer diagnosis as they allow high risk patient identication, for whom prognosis can be further improved by an adjuvant therapy. Ki67 proliferative index is a potential marker for predicting the responsiveness to chemotherapy. This present study has been designed to analyze the relationship of Ki67 proliferative index with clinico-pathological factors in eighty diagnosed cases of primary breast carcinoma, according to the molecular subtypes. To compare the e Objective: xpression of Ki67 proliferative index with the molecular subtype, histopathological grade and type of breast carcinoma. The study w Material and Methods: as carried out from October 2019 - September 2021 in the Department of Pathology, M.K.C.G Medical College & Hospital, Odisha. 80 cases received for routine histopathological examination were evaluated for histopathological typing and IHC status – ER, PR, Her2neu & Ki67. 46 cases belonged to T2 followed by Result: T3 and T1. 45% cases were Grade III followed by Grade II and Grade I tumors. 74 cases were IDC NOS, 2 cases Lobular, 2 cases Metaplastic, 1 case Medullary and 1 case Mucinous. High Ki67 index seen in 70.37% cases in T3 category followed by T2 (45.65%) and T1(42.86%). Low Ki67 index is associated with Luminal A followed by Luminal B. High Ki67 index is seen in 83.33% of Grade III tumor but none in Grade I tumor. Whereas, high Ki67 index is found in both Her2neu enriched and Triple Negative breast carcinoma. High Ki67 proliferative index Conclusion: was associated with high grade tumor, Her2/neu enriched and Triple negative tumor. However, there was no signicant relation found in association of ER, PR, Her2/neu and Ki67 with the tumor size

Comparison of GATA-3, Mammaglobin and GCDFP-15 Expression in Primary, Metastatic and Triple Negative Breast Carcinomas: An Indian Scenario.

Mammaglobin and GCDFP-15 are traditional immunohistochemistry (IHC) markers utilized to recognize metastasis of breast carcinoma in an unknown primary. GATA-3 is increasingly being used as a marker of primary breast origin. This study was done to evaluate and compare GATA-3 with GCDFP-15 and Mammaglobin in invasive primary including metastatic and triple negative breast carcinomas. Immunohistochemistry for GATA-3, GCDFP-15 and Mammaglobin was applied on 100 cases of primary breast carcinomas, including 20 triple negative cases and 30 cases of metastatic breast carcinomas. Staining scores were given for each marker by multiplying the percentage of positive tumor cells by the intensity of staining (1+, 2+ or 3+), with scores ranging from 0 to 300. Staining score of 1 or more was considered positive. GATA-3 was expressed in 92% of primary, 80% of metastatic and 60% of triple negative breast carcinomas, with an average staining score of 270. Mammaglobin was expressed in 68% of primary, 56.6% of metastatic and 25% of triple negative breast carcinomas, with an average staining score of 180. GCDFP-15 was expressed in 48% of primary, 26.6% of metastatic and 05% of breast carcinomas, with an average staining score of 60. GATA-3 demonstrated to have higher staining score (average of 270) than other two markers in maximum number of cases. GATA-3 has a higher sensitivity and increased staining scores in primary breast carcinomas, metastatic breast carcinomas as well as in triple negative breast carcinomas.

Correlation of proliferative index with various clinicopathologic prognostic parameters in breast carcinoma

Introduction: Breast cancer (BC) is the most common cancer and the second most common cause of death in women. Prognostic factors are essential in BC diagnosis as they allow the identification of high-risk patients, for whom, an adjuvant therapy can improve prognosis. Interest in Ki-67 has recently increased as Ki-67 is a potential marker for predicting the responsiveness to chemotherapy. Materials and Methods: The study was conducted on 75 consecutive cases of primary breast carcinoma undergoing radical or modified radical mastectomy specimen. Histopathological diagnosis was established on routine hematoxylin and eosin stain and various histologic prognostic parameters including histologic type, histologic grade, and lymph node metastases were assessed. All biopsy proven carcinoma breast were included in this study. Results: Among presenting compliant, 47 (62.66%) had lump in right breast and 28 (37.33%) had lump in left breast. Most patients in the study population presented with a lump of duration 3-6 months (54.6%). Out of 75 participants, 41 (62.66%) participants were diagnosed with carcinoma right beast and 28 (37.33%) with carcinoma left breast. On staging the disease among the study population, most of them diagnosed with breast cancer were of stage IIIB (48%) followed by IIA (25.33%).

A Study of Tumour Infiltrating Lymphocytes (Tils) In Breast Carcinoma and their Immuno-Histochemical Profile

Objective: Tumor-infiltrating lymphocytes (TILs) play an important role in mediating immune response against cancer cells and are associated with improved clinical outcomes. The present study was conducted to demonstrate the relative densities of T lymphocytes, CD4+ cells, CD8+ cells, and B-lymphocytes in a series of breast carcinoma. Material and method: Thirty cases of primary breast carcinoma were examine for the histological parameters including histological type, histological grade and lymph node metastases and pattern of inflammatory reaction. Further, immunohistochemical expression in cases with lymphocytic infiltrate was studied in order to classify various lymphocyte subsets. Results: T-cells (CD3+) were present in 96.6% cases of breast cancer, majority of which were CD8+ cells in 90% cases and CD4+ cells in 36.6% cases. B-cells were present in 83.3% cases. Also, insignificant correlation was found between TILs and subtypes with age, menopausal status, histological grades, ER, PR and HER2/neu status and tumor size. No significant correlation was found between CD4, CD8 and CD20 with lymph nodes stage in our study. However, significant correlation was found between CD3 with lymph nodes stage. Conclusion: T lymphocytes and their subsets (especially CD8+ cells) predominated over B lymphocytes quantitatively and qualitatively. They seem to promote neoplastic progression rather than acting as a protective immune response against cancer

Isolation and morphology of circulating tumor cells by cell block technique in breast cancer.

Circulating tumor cells (CTCs) are cells present in the blood stream that are antigenically or genetically similar to a specific tumor type and are markers of tumor diagnosis, prognosis, residual disease and metastasis. The ever-increasing burden of breast cancer globally warrants the incorporation of this all-inclusive marker in the diagnostic repertoire using the simplest of techniques. To identify CTCs in peripheral blood by cell block (CB) technique in cases of breast cancer diagnosed on fine-needle aspiration (FNA) or core needle biopsy (CNB) and to correlate their presence with nodal metastasis. This study was conducted in the Department of Pathology, at a tertiary care hospital. Peripheral blood samples from a total of 30 cases of primary breast carcinoma diagnosed on FNA or CNB without prior neoadjuvant chemotherapy were analyzed using the CB technique. The age ranged between 29-74 years with the most common presenting complaint being a palpable, single, unilateral breast lump. CTCs were detected in 2 (6.7%) cases with a <5 cell cluster with both the cases being grade I breast carcinomas and also displaying nodal metastasis.

Study of ER, PR, HER2/neu, p53, and Ki67 expression in primary breast carcinomas and synchronous metastatic axillary lymph nodes.

Breast cancer (BCA) is the second most common cancer among women in India and accounts for 7% of global burden of BCA. The axillary lymph node status is an independent prognostic factor. The combined estrogen receptor (ER), progesterone receptor (PR), and HER2/neu biomarker expression is a predictor of BCA status for therapeutic guidance. Studies have demonstrated that these biomarkers are unstable throughout their tumor progression. Varying concordance and discordance rates in the biomarker expression between primary breast carcinoma (PBC) and metastatic axillary lymph node (MALN) status are reported. This study was conducted for studying and comparing the expression of immunohistochemistry (IHC) markers, i.e., ER, PR, HER2/neu, p53, and Ki67 between PBC and their corresponding MALN for prognostication and therapeutic purpose. Sixty cases of PBC with metastasis to axillary lymph nodes diagnosed between years 2008 and 2014 were included in the study. A technique of manual tissue array was employed for cases subjected to IHC. Analyses of the expression of IHC markers were attempted between the PBC and their corresponding synchronous MALN and classified as concordant or discordant. Results were subjected to statistical analysis. Substantial agreement was observed for biomarker ER, PR, HER2/neu, p53, and Ki67 expression between PBC and MALN with k-value 0.79, 0.75, 0.89, 0.7, and 0.6, respectively. There was high concordance for the IHC markers: ER, PR, HER2/neu, p53, and Ki67 expression in matched pairs of PBC and corresponding synchronous MALN. However, the discordance noted in small subgroups cannot be overlooked. Thus, there is a need to perform ER, PR, HER2/neu, p53, and Ki67 IHC studies routinely in both PBC and MALN to help design therapies that are tailored to target the specific tumor clones and render maximum benefit to patients.

Effects of Hydrochloric Acid and Formic Acid Decalcification on Breast Tumor Biomarkers and HER2 Fluorescence In Situ Hybridization.

Biomarker analysis of metastatic breast carcinoma (MBC) is routinely recommended by ASCO/CAP guidelines, and establishing a diagnosis of MBC often requires immunohistochemistry (IHC). The reliability of breast tumor biomarkers and breast-specific markers on decalcified tissues has not been extensively studied. We performed IHC studies on breast tumors exposed to hydrochloric acid (HCl) and formic acid (FA) decalcification solutions, and HER2 fluorescence in situ hybridization on a subset of these tumors to establish a protocol for handling bone specimens with suspicion for MBC. Fifteen fresh cases of primary breast carcinoma and 8 HER2+ paraffin-embedded core biopsy cases were studied. Fresh tissue was divided into 5 fragments to approximate a bone core biopsy. One fragment (control) was fixed in 10% neutral buffered formalin. The remaining fragments were also exposed to FA or HCl decalcification for 1 or 5 hours. All fragments were embedded in 1 block and tested with an IHC panel. The known HER2+ cases were exposed to either 1 or 5 hours of FA, and HER2 fluorescence in situ hybridization was also performed. Results were interpreted as follows: H-scores for estrogen receptor, progesterone receptor, and GATA-3 were assigned from 0 to 300; HER2, cytokeratin 7, gross cystic disease fluid protein-15, Pax-8, TTF-1, cytokeratin 20, and mammaglobin were scored from 0 to 3+; and Ki67 from 0% to 100%. Mean scores were compared using the t test or Wilcoxon test for paired samples. No significant differences in mean score were seen between NF and 1 hour FA for any IHC immunoreactivity. After 5 hours of FA, only Ki67 average score was significantly less than NF. Mean scores for estrogen receptor, progesterone receptor, HER2, Ki67, and GATA-3 were significantly lower than NF in the tissue after either 1 or 5 hours of HCl. Mean scores for gross cystic disease fluid protein-15, mammaglobin, and cytokeratin 7 staining were not significantly lower than NF after 1 or 5 hours of HCl.

Hormone Receptors and Human Epidermal Growth Factor (HER2) Expression in Fine-Needle Aspirates from Metastatic Breast Carcinoma - Role in Patient Management.

Introduction:Estrogen receptors (ER), progesterone receptors (PR), and epidermal growth factor (HER2) are prognostic and predictive factors for breast carcinoma. We determined them by immunohistochemistry (IHC) on cell blocks from fine-needle aspirates (FNA) of metastatic breast carcinoma to axillary lymphnodes and compared them with that reported in the primary breast carcinoma (PBC) to document any change in their expression for future management.Materials and Methods:ER, PR, and HER2 by IHC and HER2 oncogene by fluorescent in-situ hybridization (FISH) were studied on cell blocks of FNA of axillary lymphnodes in 53 of 94 PBC cases from 2012 to 2016.Results:In 25 of 38 (65.8%) ER, PR negative PBC the metastasis on FNA was ER, PR+, whereas the 15 (28.3%) ER, PRPBC remained negative. In 10 of 11 (91%) of HER2-IHC+, PBC the metastatic tumor was HER2-IHC+. 7 of 32 (21.9%) HER2-IHC negative PBC were HER2-IHC+ in metastatic tumor. HER2-FISH was performed in 37 cases on FNA. Six of 37 were HER2 amplified/positive, whereas 9 and 19 remained equivocal and negative for HER2 copy number, and 3 were not interpretable. All the 6 HER2-FISH+ cases were positive by IHC. In our study, 34.2% of ER, PR+ cases of PBC became ER, PR– in the metastatic tumor and 21.9% of HER2-IHC negative PBC became HER2-IHC+ in the metastatic aspirate.Conclusion:ER, PR, and HER2 by IHC in cell blocks of metastatic lymphnodes are reliable. Change in receptor (34.2%) and HER2 status (21.9%) was documented, which is of clinical significance as these patients warrant a change of management.

AKT signaling pathway in invasive ductal carcinoma of the breast: correlation with Erα, ERβ and HER-2 expression

Estradiol exerts most of its effects by direct binding to the estrogen receptor in breast carcinoma, ERβ expression is a useful biomarker for breast cancer in a manner that is independent of ERa expression. However, studies evaluating ERβ expression with certain tumor variables, such as tumor grade and disease-free survival, had produced conflicting results. The Akt signaling pathway currently attracts considerable attention as a new target for effective therapeutic strategies. The current study attempted to compare the relative associations of variables including ERa, ERβ, HER-2/neu and AKT staining with the presence of metastases or survival. Immunohistochemical staining was employed to determine the expression of ERa, ERβ, pAkt and HER-2/neu in 110 cases of primary breast carcinoma. Positive ERa, ERβ, pAkt and HER-2/neu expressions were respectively observed in 46.4% (51/110), 59.1% (65/110), 40.9% (45/110) and 31.8% (35/110) of the tumors. pAkt was significantly associated with HER-2/neu overexpression (P <0.005) and axillary lymph node metastasis (P <0.05). However, there was no significant relationship between pAkt and ERa, ERβ, p53 (P >0.05) expressions. Survival analysis showed that pAkt positivity was associated with poor disease-free survival of the patients. The current study suggested that activity of the Akt signaling pathway may indicate a poor prognosis in patients with breast carcinoma. The results implied that estrogen can activate the PI3K-Akt pathway through ERa and ERβ-independent mechanisms in breast cancer.

Immunohistochemical Study of MUC1, MUC2 and MUC5AC Expression in Primary Breast Carcinoma.

Breast Cancer (BC) is the second most common cancer among women in India and accounts for 7% of global burden of BC and one-fifth of all Cancers (CA) among women in India. This study was conducted for studying the expression of MUC1, MUC2 and MUC5AC in breast carcinoma. Fifty cases of primary breast carcinoma diagnosed between years 2013 to 2015 were included in the study. Manual tissue array technique was applied for cases subjected to Immunohistochemistry (IHC). An analysis of the expression of IHC markers (MUC1, MUC2, MUC5AC, ER, PR and HER2/neu) was attempted. Results were subjected to statistical analysis. They were considered to be significant when the p-value was less than 0.05. The positivity for MUC1, MUC2 and MUC5AC in BC was 58%, 8% and 6% and for ER, PR and HER2 was 48%, 36% and 64% respectively. There was a significant correlation between MUC1 expression and ER and PR positivity. There was a significant correlation between MUC2 expression and ER positivity. No significant association was observed between MUC2 and PR expression, MUC5AC expression and ER and PR positivity. There was statistically significant correlation between negative MUC2 and MUC5AC expression and histopathological grade. It was noted that MUC2 and MUC5AC negative tumours were associated with higher tumour stage though not statistically significant. It was noted that MUC5AC negative tumours showed higher frequency of lymphovascular invasion though not statistically significant. Our experience with the present study highlights the role of mucins in the development and progression of BC.

Folate Receptor Alpha Immunohistochemistry in Cytology Specimens of Metastatic Breast Carcinoma

Background: Folate receptor alpha (FRA) is involved in folate accumulation and utilization, and is expressed in varying proportions in breast, ovary and parotid epithelial cells, among others. FRA overexpression by immunohistochemistry (IHC) has been shown in estrogen/progesterone receptor (ER/PR)-negative carcinoma (40-74%) and in triple-negative breast carcinoma (TNBC; 50-86%) in histological specimens of primary breast cancers. We assessed the feasibility of IHC in detecting FRA expression and its patterns and clinical significance in metastatic TNBC in fine-needle aspiration (FNA) cell blocks (CBs). Materials and Methods: Metastatic breast ductal carcinoma cases were retrospectively immunostained with FRA IHC on FNA CBs. FRA staining was scored qualitatively (+/-), by intensity (0-3) and by staining area (0-100%). Of these metastatic cases, a subset of primary breast carcinoma cases was also immunostained with FRA. The results were correlated with ER, PR and human epidermal growth factor receptor 2 (Her2/Neu) performed by routine IHC. Results: A total of 40 FNA CBs with metastatic disease were studied, including hormone (ER/PR) positive (n = 5), triple positive (n = 5), Her2/Neu-only positive (n = 5) and TNBC (n = 25). FRA IHC showed immunoreactivity with moderate positivity in only 1 (4%) TNBC. All the remaining 39 cases were negative for FRA expression. Five cases of primary TNBC were stained with FRA IHC and were negative for FRA expression. Conclusions: Our data suggest that FRA expression by IHC was rarely associated with ER/PR-negative tumors relative to ER/PR-positive tumors and, more importantly, with TNBC in FNA CBs. This finding may have a clinical significance and prognostic implications in metastatic breast carcinoma. Furthermore, 5 primary TNBC cases did not overexpress FRA by IHC. Hence, antifolate receptor therapies do not appear to be clinically relevant in TNBC based on immunostaining of FNA CBs of metastatic breast cancers.

AXILLARY LYMPH NODE STATUS IN PRIMARY BREAST CARCINOMA

Objectives: The most important prognostic factor in patients of breast carcinomais axillary lymph node metastasis. Current study was conducted to find the frequency of lymphnode metastasis in hundred cases of primary breast carcinoma and association of lymphnode status with immunohistochemical expression of ER/PR,HER2/neu and MMP-1(matrixmetalloproteinase-1).Design: Descriptive study. Period: Aug 2012 to Jun 2013. Setting: U.H.Slaboratory of Morbid Anatomy and Histopathology Lahore. Materials and methods:Onehundred mastectomy specimens with axillary lymph node dissection were included. Aftergross examination, tissue processing and microtomy the tissue slices of 4 micrometer weretaken on frosted and lysine coated slides. H/E and IHC for ER/PR,HER2/neu and MMP-1 weredone according to protocol. Results: Among 100 breast cancer subjects, 72 were positivefor lymph node metastasis while 28 subjects were negative. A significant association betweenlymph node status and ER IHC expression was noticed. When Chi square test was appliedwith p-value of 0.001 was observed.Also a significant association between lymph node statusand PR IHC expression was noticed. Chi square test was applied and the p-value of 0.004 wasobtained.Association between lymph node status and HER2/neu IHC expression was analysed.Chi square test was applied with a p-value of 0.467 was obtained (not significant).A significantassociation between lymph node status and MMP-1 IHC expression was observed. Chi squaretest was applied anda p-value of 0.004 was obtained. Conclusions: Most of the primary breastcarcinomas were presented with axillary lymph node metastasis (72%).Significant associationswere observed between lymph node status and ER/PR immuonohistochemical expressionshowever the association between HER2/neu IHC and lymph node status was not statisticallysignificant. The immunohistochemical expression of MMP-1 is significantly associated withlymph node status. It shows that it is an important marker for metastatic potential in breastcarcinoma.

Correlation of various histopathologic prognostic factors with Nottingham prognostic index and microvessel density in invasive breast carcinoma: A study of 100 cases.

Nottingham prognostic index (NPI) is a widely used integrated prognostic variable in patients with breast cancer. NPI has been correlated with tumor size, grade, lymph node stage and patient survival. The present study aimed at evaluating and correlating the various clinical and pathologic features of breast carcinoma with NPI. This study included 100 consecutive cases of primary breast carcinoma over a period of 2 years. Demographic data was noted and histomorphological features like tumor size, grade, lymph node involvement, necrosis, vascular invasion etc., were assessed. NPI was calculated as reported in the literature. Immunohistochemical staining for hormone receptors and CD34 (to calculate microvessel density [MVD]) was performed. Statistical analysis was used for correlation. Of the 100 cases, 54% of the tumors were in T2 tumor size category (2-5 cm) and lymph node metastasis in 48% of the cases. NPI ranged from 2.3 to 7.3 with 54% of the cases in the intermediate NPI group (3.41-5.4). The mean MVD was 160.93 (±69.4/mm2). On statistical analysis, tumor size and grade, lymph node stage, mitotic rate, nuclear pleomorphism, necrosis and MVD showed a correlation with NPI (P < 0.05). NPI is an important and useful prognostic indicator for breast cancer patients, which shows the correlation with other histomorphological prognostic features as well.

The Utility of Toluidine Blue for Assessing Cross Contamination in Fluid Specimens

Results: All of the patients found in this study presented with malignant pleural effusion or ascitesdother serous cavity effusions resulting from metastatic breast carcinoma were not seen. “Cannonball” formation was only found in 5 of the six cases of moderately differentiated (Nottingham grade 2) breast carcinoma (Figure 1). All five of these cases had Nottingham sub-scores of 3 for tubule formation, 2 for pleomorphism, and 1 for mitotic activity. The 5 cases of primary breast carcinoma with poor differentiation (grade 3) and with malignant effusion all displayed single malignant cells or dyshesive clusters of malignant cells (Figure 2). The average time from breast biopsy to detection of malignant effusion was 298 days for grade 2 primary breast carcinoma and 2003 days for grade 3 primary breast carcinoma (Table 1). Immunohistochemistry for common breast carcinoma molecular markers was noted for the original biopsy and malignant effusion. In the malignant effusion, ER and PR were often lost while mammoglobin was commonly retained (Table 2).

Clinicopathological features and prognostic factors of young breast cancers in Eastern Guangdong of China.

Breast cancer in young women is typically with higher proportion of adverse pathological features. Breast cancer with BRCA1 mutation is often early-onset, and is usually associated with triple negative phenotpe. In this study, we aim to analyze the clinicopathological characteristics and prognosis in young breast cancer patients (≤35 years old) comparing to non-young patients (>35 years old). A total of 1913 cases of primary breast carcinoma with stage I–III were enrolled, with 283 cases diagnosed as young patients. No significant difference was observed in tumor size, TNM staging, lymph node metastasis, ER, HER-2 or histological grade between young and non-young patients. Multivariate analysis demonstrated that age was an independent prognostic factor for overall survival (OS). In 70 samples of young patients available, BRCA1 was immunohistochemically positive 85.7% in cytoplasm and 41.4% in nuclear. BRCA1 nuclear expression is not significantly associated with clinicopathological characteristics in young breast cancer patients.

Ethnic disparities in breast cancer between Central Europe Caucasian women of Slavic origin and Middle East Turkish subjects

The biological, cultural, behavioral and sociodemographic differences across populations modulate breast cancer profile among races or ethnics. Following this, we aimed to identify differences in breast cancer epidemiology, histopathology, and clinical presentation from representatives of central Europe (Slovakia) and Middle-East countries (Turkey) to point on ethnic disparities in cancer biology. The population based cross-sectional study analyzing 414 cases of primary breast carcinomas where 214 represented Caucasian and 200 Turkish subjects. The differences were found for age at the time of diagnosis (<0.0001), education, menopausal status (<0.001), tumor localization (<0.01), size (<0.0001), grade (<0.05) and axillary lymph node status (<0.001) between groups. Although carcinomas in Slovak subjects were of higher grade, negative axillary nodal status was more frequent finding compared to Turkish patients (50.0 vs. 41.0%). The Slovak group showed carcinomas to be more often ER positive (72.4 vs. 54.0%; <0.001), ER/PgR positive (54.6 vs. 49.0%; <0.001), of better Nottingham prognostic index (<0.001), and less frequent Her-2 positive (21.2 vs. 28.5%). Slovak population expressed significantly higher risk of non-sentinel lymph node metastases with increased tumor size, grade, vascular invasion and Her-2 positivity compared to Turkey population. The tumor size >2 cm and high tumor grade (G3) bears a risk of OR=7.62 and OR=3.10 in Slovak compared to OR=3.94 and OR=1.79 in Turkish cases, respectively.There are wide demographic and biological disparities in breast cancer between observed ethnics providing unique information for clinicians working at the level of screening or therapy in these populations.

In Situ Identification of CD44+/CD24− Cancer Cells in Primary Human Breast Carcinomas

Breast cancer cells with the CD44+/CD24− phenotype have been reported to be tumourigenic due to their enhanced capacity for cancer development and their self-renewal potential. The identification of human tumourigenic breast cancer cells in surgical samples has recently received increased attention due to the implications for prognosis and treatment, although limitations exist in the interpretation of these studies. To better identify the CD44+/CD24− cells in routine surgical specimens, 56 primary breast carcinoma cases were analysed by immunofluorescence and confocal microscopy, and the results were compared using flow cytometry analysis to correlate the amount and distribution of the CD44+/CD24− population with clinicopathological features. Using these methods, we showed that the breast carcinoma cells displayed four distinct sub-populations based on the expression pattern of CD44 and CD24. The CD44+/CD24− cells were found in 91% of breast tumours and constituted an average of 6.12% (range, 0.11%–21.23%) of the tumour. A strong correlation was found between the percentage of CD44+/CD24− cells in primary tumours and distant metastasis development (p = 0.0001); in addition, there was an inverse significant association with ER and PGR status (p = 0.002 and p = 0.001, respectively). No relationship was evident with tumour size (T) and regional lymph node (N) status, differentiation grade, proliferative index or HER2 status. In a multivariate analysis, the percentage of CD44+/CD24− cancer cells was an independent factor related to metastasis development (p = 0.004). Our results indicate that confocal analysis of fluorescence-labelled breast cancer samples obtained at surgery is a reliable method to identify the CD44+/CD24− tumourigenic cell population, allowing for the stratification of breast cancer patients into two groups with substantially different relapse rates on the basis of CD44+/CD24− cell percentage.

Determination of human epidermal growth factor receptor 2 (HER2) amplification using fluorescence in situ hybridization (FISH), chromogenic in situ hybridization (CISH) and automated silver in situ hybridization (SISH)

Human epidermal growth factor receptor 2 (HER2) amplification is an important biomarker in breast carcinoma and its determination has been validated in the past using fluorescence in situ hybridization (FISH). However, HER2 FISH method requires specialized fluorescence microscope, high cost and impermanent signals causing it to be impractical for routine laboratories. The aim of the present study was to determine the expression of HER2 using FISH and another two recent new techniques, such as chromogenic in situ hybridization (CISH) and silver in situhybridization (SISH). The whole sections of HER2 of 25 primary breast carcinoma cases with borderline positive (2+) immunohistochemistry (IHC) were further validated by using manual dual-color FISH, manual single color CISH and automated single color SISH. A total of 88, 84 and 92% of the cases showed HER2 amplification using FISH, CISH and SISH, respectively. A high level of concordance between FISH and CISH, FISH and SISH, CISH and SISH were observed in 91.6% (p = 0.032, k = 0.619), 96% (p = 0.01, k = 0.779) and 95.8% (p = 0.011, k = 0.778), respectively. SISH technique saved time, CISH equipment was less expensive and their high level of concordance shows that they may be alternatively used in routine laboratories. Key words: Breast carcinoma, HER2, fluorescence in situ hybridization (FISH),chromogenic in situ hybridization (CISH), silver in situ hybridization (SISH), immunohistochemistry.