Breast Surgical Specimens Research Articles

Northwestern profiling of potential translation-regulatory proteins in human breast epithelial cells and malignant breast tissues: evidence for pathological activation of the IGF1R IRES

Genes involved in the control of cell proliferation and survival (those genes most important to cancer pathogenesis) are often specifically regulated at the translational level, through RNA-protein interactions involving the 5'-untranslated region of the mRNA. IGF1R is a proto-oncogene strongly implicated in human breast cancer, promoting survival and proliferation of tumor cells, as well as metastasis and chemoresistance. Our lab has focused on the molecular mechanisms regulating IGF1R expression at the translational level. We previously discovered an internal ribosome entry site (IRES) within the 5'-untranslated region of the human IGF1R mRNA, and identified and functionally characterized two individual RNA-binding proteins, HuR and hnRNP C, which bind the IGF1R 5'-UTR and differentially regulate IRES activity. Here we have developed and implemented a high-resolution northwestern profiling strategy to characterize, as a group, the full spectrum of sequence-specific RNA-binding proteins potentially regulating IGF1R translational efficiency through interaction with the 5'-untranslated sequence. The putative IGF1R IRES trans-activating factors (ITAFs) are a heterogeneous group of RNA-binding proteins including hnRNPs originating in the nucleus as well as factors tightly associated with ribosomes in the cytoplasm. The IGF1R ITAFs can be categorized into three distinct groups: (a) high molecular weight external ITAFs, which likely modulate the overall conformation of the 5'-untranslated region of the IGF1R mRNA and thereby the accessibility of the core functional IRES; (b) low molecular weight external ITAFs, which may function as general chaperones to unwind the RNA, and (c) internal ITAFs which may directly facilitate or inhibit the fundamental process of ribosome recruitment to the IRES. We observe dramatic changes in the northwestern profile of non-malignant breast cells downregulating IGF1R expression in association with acinar differentiation in 3-D culture. Most importantly, we are able to assess the RNA-binding activities of these putative translation-regulatory proteins in primary human breast surgical specimens, and begin to discern positive correlations between individual ITAFs and the malignant phenotype. Together with our previous findings, these new data provide further evidence that pathological dysregulation of IGF1R translational control may contribute to development and progression of human breast cancer, and breast metastasis in particular.

Az emlődaganatok primer szisztémás kemoterápiára adott válasza az immunhisztokémiai fenotípus tükrében

The purpose of the study was to identify breast cancer subtypes by immunohistochemistry likely to respond to neoadjuvant chemotherapy and to analyze the used chemotherapy regimen and the range of response rates. Analysis of a collected database was performed. Ninety-two patients were identified in our files who received neoadjuvant chemotherapy between 1998 and 2009. We used immunohistochemical profiles (ER, PgR, HER2, Ki-67 and p53) of NCB, FNAB and surgical breast specimens to subclassify the tumors. Pathological response rates were assessed following surgical removal of tumors by using the Chevallier classification. DFS and OS was measured in 88 cases from the date of definitive surgery to the date of last follow-up or death. Pathological complete or near-complete remission (pCR = Chevallier I and II) was observed in 13 of 92 cases (14.1%). According to the preoperative characteristics of the 13 tumors achieving pCR, 9 of the cases were triple negative, one of 13 was ER-/HER2+ and three of 13 ER+/HER2+. Twenty-four of 92 patients received taxane based neoadjuvant chemotherapy, 30 of 92 anthracycline based neoadjuvant chemotherapy, 33 of 92 taxane + anthracycline regimen and 2 of 92 CMF regimen. In the taxane treated group of patients the pCR rate was 29.1%, in the anthracycline group 6.6% and in the taxane + anthracycline treated group 12.1%. Concerning DFS, significant difference was observed between the Chevallier III and IV groups (p=0.006), and less events were observed in the pCR group (not significant). pCR was associated with significantly better OS (p=0.050). It seems that even limited, routinely used immunohistochemical profiling of tumors is able to predict the likelihood of pCR to neoadjuvant chemotherapy. Patients with triple negative and HER2-positive cancers are likely to achieve pCR after neoadjuvant chemotherapy.

Application of the Probabilistic Approach to Reporting Breast Fine Needle Aspiration in Males

To apply the probabilistic approach to a series offine needle aspiration (FNA) samples of male breast lesions and determine the accuracy and reproducibility of this method of reporting in men.All male breast surgical specimens with a preoperative breast FNA at our institution from 1994 to 2005 were identified. The FNAs were blindly reviewed by 2 groups of observers and classified in 1 of 5 categories using published reporting guidelines: positive, suspicious, atypical, proliferative without atypia and unremarkable. The histologic and cytologic diagnoses were correlated. The interobserver variation was determined.A total of 138 FNAs were performed for 123 male patients. Histologic correlation was available for 23 satisfactory FNAs. A total of 11 of 11 carcinomas (100%) were classified as positive, suspicious or atypical. Of 12 benign masses, 11 (91.6%) were classified as proliferative without atypia or unremarkable. One case of gynecomastia was classified as atypical by 1 observer but deemed not atypical with consensus review. The kappa statistic for benign and atypical/suspicious/malignant categories was 0.90.Based on this series, the probabilistic approach can be applied to the reporting of FNAs of male breast lesions. Gynecomastia may result in an atypical cytologic diagnosis.

Histogram partition and interval thresholding for volumetric breast tissue segmentation

It is possible to automatically decompose a volume into subvolumes based on histogram partition and interval thresholding. In practice, a histogram may assume unimodal or multimodal distributions. In this paper, we implement an automatic volumetric segmentation scheme by partitioning a histogram into intervals followed by interval thresholding. Based on its distribution shape, the histogram is partitioned by either a valley-seeking algorithm (for multimodal) or a five-subinterval algorithm (for unimodal). Applied to volumetric breast analysis, this technique decomposes a breast volume into five subvolumes corresponding to five intensity subintervals: lower (air bubble), low (fat), middle (normal tissue, or parenchyma), high (glandular duct), higher (calcification), in the order of X-ray attenuation value. With the assumption that each subvolume resulting from interval thresholding corresponds to a tissue type, the spatial structure of each breast tissue type can be individually visualized and analyzed in a subvolume in an ample space (as big as the whole volume) in the absence of other tissue types. We demonstrate this histogram-partitioned interval thresholding segmentation method with one breast phantom and one breast surgical specimen that are volumetrically reconstructed by cone-beam tomography.

Guidelines for the handling of benign and malignant surgical breast specimens

Summary Most histopathology laboratories routinely handle diagnostic and excisional breast surgical specimens, and the range of specimens received is limited. However, the apparent simplicity of breast specimens is often deceptive, and the importance of thorough macroscopic assessment is often underestimated. Surgical techniques vary between institutions, and an understanding of the local surgical protocol is essential for optimal specimen handling. Adequate and appropriate sampling is essential for optimal reporting of the minimum data set information, particularly an accurate assessment of the size of the lesion and the excision margin status. This article offers some guidelines for the handling of benign and malignant surgical breast specimens, including more complex specimens such as excisions following neo-adjuvant therapy.

Hiperplasias epiteliais em espécimes de mamoplastia redutora estética bilateral e mamoplastia redutora contralateral a câncer de mama

INTRODUCAO: A mamoplastia redutora estetica bilateral (MEB) e uma cirurgia que tem como objetivo reduzir o volume mamario, melhorando problemas esteticos e posturais, ou obter equilibrio estetico em pacientes submetidas a mamoplastia contralateral (MRC) a câncer de mama. A incidencia de lesoes mamarias nestes especimes e variavel, e a presenca de câncer em uma das mamas e fator de risco conhecido para desenvolvimento de câncer na mama contralateral. OBJETIVOS: Comparar a frequencia de lesoes proliferativas epiteliais e outras alteracoes histopatologicas em especimes de MEB e MRC. MATERIAIS E METODOS: Foram revistas lâminas de 867 pecas cirurgicas de MEB e 72 de MRC, anotando-se as alteracoes histopatologicas, com enfase nas hiperplasias epiteliais, e relacionando-as com a faixa etaria das pacientes. RESULTADOS: A idade media das pacientes submetidas a MEB foi de 34 anos, e das submetidas a MRC, de 50,3 anos. Alteracoes fibrocisticas foram as lesoes mais comumente diagnosticadas. Cistos e ectasia ductal estavam presentes em 50,9% das MEB e em 59,7% das MRC. Hiperplasias epiteliais foram diagnosticadas em 10,1% das MEB e em 43,1% das MRC, principalmente em mulheres com idade superior a 40 anos. Entre as hiperplasias, o tipo usual foi o mais frequente nos dois grupos (MEB = 9,8% e MRC = 33,3%). Hiperplasias atipicas estavam presentes em 9,7% dos casos de MRC e em 0,3% de MEB. CONCLUSAO: A incidencia de hiperplasias epiteliais com e sem atipias foi maior nos casos de MRC do que nos de MEB mesmo quando as pacientes foram pareadas pela faixa etaria.

Intramammary lymph node metastases are an independent predictor of poor outcome in patients with breast carcinoma.

Breast carcinoma with intramammary lymph node (intraMLN) metastases is considered to be Stage II disease, even in the absence of axillary lymph node involvement. Nonetheless, little is known regarding the clinical significance of intraMLN metastases. The goals of the current retrospective analysis were to elucidate the clinical relevance of intraMLN metastases and to assess the relation between such metastases and outcome in patients with breast carcinoma. One hundred ninety-six intraMLN specimens obtained between 1983 and 2003 were identified in the pathology database at The University of Texas M. D. Anderson Cancer Center (Houston, TX); 130 of these specimens were obtained in association with a primary breast malignancy. Data on the clinical and pathologic features of these specimens were collected and evaluated on univariate and multivariate analysis for potential correlations with 5-year rates of disease-free survival (DFS), disease-specific survival (DSS), and overall survival (OS). The median follow-up duration was 36 months (range, 12-180 months). The median age of the 130 patients in the current study was 53 years (range, 27-84 years). Twenty-four patients (18%) had intraMLNs that were identified preoperatively by either mammographic or sonographic methods; in the remaining 106 cases, intraMLNs were detected on pathologic examination of surgical breast specimens. IntraMLN metastases were found in 28% of all cases (n = 36). Most patients who had intraMLN metastases (81%) also had axillary metastases; however, isolated intraMLN metastases were documented in 6 patients (5%). Univariate analysis revealed that patients with intraMLN metastases (compared with all other patients) had poorer 5-year rates of DFS (54% vs. 89%; P = 0.001), DSS (66% vs. 90%; P = 0.001), and OS (64% vs. 88%; P = 0.004). Furthermore, multivariate analysis indicated that intraMLN involvement was an independent predictor of reduced DFS (hazard ratio, 2.33; P = 0.03), DSS (hazard ratio, 5.32; P = 0.002), and OS (hazard ratio, 3.22; P = 0.006). The current retrospective analysis demonstrated that the presence of intraMLN metastases is an independent predictor of poor outcome in patients with breast carcinoma. Identification of an intraMLN on preoperative imaging should prompt further histopathologic assessment. Identification of malignant intraMLNs by lymphatic mapping may help to identify high-risk patients for whom further evaluation of the axillary lymph nodes is warranted despite otherwise clinically negative findings in the axilla.

Accuracy and Consistency in Application of a Probabilistic Approach to Reporting Breast Fine Needle Aspiration

To determine the application of a probabilistic/categorical approach for reporting breast fine needle aspiration (FNA) and its dependence on the cytopathologist's level of experience. All breast surgical specimens that had preoperative breast FNA at our institution during a 3-year period were identified. The cytologic results were reported as 1 of 6 categories: positive, suspicious, atypical, epithelial proliferative, unremarkable and nondiagnostic, according to well-defined criteria. Five cytopathologists were responsible for all cytology sing-out during the study period. The histologic and cytologic diagnoses were correlated. A total of 297 cases were identified. Overall, there were no false positive cases (positive predictive value [PPV] = 100%). Two false negative cases (negative predictive value [NPV] = 96%) were due to sampling error. This indicates that the PPV and NPV for each of the 5 pathologists were also all 100% except for the 1 pathologist who had two false negative cases due to sampling errors. The probability of finding carcinoma on histology for suspicious and atypical cytologic categories ranged from 67% to 100% and 8% to 31%, respectively, for the individual pathologists. Fifteen cases were signed out by > or = 2 pathologists. The involvement of consultants was significantly associated with diagnosis (P = .02). Ten of the 15 cases were in the suspicious (5) or atypical (5) category. The probabilistic approach with defined diagnostic criteria is an accurate method and can be consistently applied in reporting breast FNA. Although use of the indeterminate (suspicious and atypical) categories is variable, a definite and considerable difference in the probability of carcinoma between these 2 categories was observed for all pathologists. The involvement of consultants did not move the cases out of these indeterminate categories.

Insulin-like growth factor system gene expression in women with type 2 diabetes and breast cancer

Background/Aims: A twofold increased risk for breast cancer has been reported recently for women with late onset diabetes. Most studies showed that there were differences in serum concentrations of insulin-like...

Why should breast tumour detection go three dimensional?

Although x-ray mammography is widely developed for breast tumour detection, it suffers from spatial superposition in its two-dimensional (2D) representation of a three-dimensional (3D) breast structure. Accordingly, 3D breast imaging, such as cone-beam computed tomography (CT), arises at the historic moment. In this paper, we theoretically elucidate the spatial superposition effect associated with x-ray mammography on breast tumour detection. This explanation is based on the line integral of x-ray traversing a composite breast model. As a result, we can characterize the difficulty of detecting small tumours in terms of local intensity contrast in x-ray images. In comparison, we also introduce cone-beam CT breast imaging for 3D breast volume representation, which offers advantages for breast mass segmentation and measurement. The discussion is demonstrated with an experiment with a breast surgical specimen. In conclusion, we strongly believe that 3D volumetric representation allows for more accurate breast tumour detection.

Comparison of Full-Field Digital Mammography to Screen-Film Mammography with Respect to Diagnostic Accuracy of Lesion Characterization in Breast Tissue Biopsy Specimens

Rationale and Objectives This study was performed to investigate whether full-field digital mammography (FFDM) is at least as accurate as screen-film mammography with respect to breast lesion characterization. Materials and Methods Seventy-nine breast surgical specimens were obtained by means of preoperative needle localization with surgical excision from 79 patients. The specimens were imaged with both screen-film mammography and FFDM. Six radiologists specialized in breast imaging analyzed both sets of images and characterized the visualized lesions on a five-point scale: 1, definitely not malignant; 2, probably not malignant; 3, possibly malignant; 4, probably malignant; and 5, definitely malignant. Receiver operating characteristic curve analysis of the data was then performed to assess for differences between modalities in the radiologists' ability to predict breast malignancy. Results The areas under the receiver operating characteristic curves for the prediction of breast malignancy in surgical biopsy specimens were not statistically significantly different for FFDM and screen-film mammography. Conclusion The results demonstrate that with breast surgical specimens, FFDM is similar in diagnostic accuracy to screen-film mammography.

Histological findings in surgical specimens after core biopsy of the breast

Pathological changes induced by needling procedures found in breast surgical specimens are rare but can induce misinterpretation or compromise the definitive histological analysis. These abnormal findings depend on the interval between the core biopsy and excision. Early findings are local haemorrhage, disrupted tissue and epithelial cell displacement, whereas, fibrosis, fat necrosis and inflammatory reaction are observed later in time. The radiologists must be aware of these histological pitfalls and must consider the benefits of their core biopsies (indications, surgeon's question, number of samples).

Histomorphologically intact primary human breast lesions and cancers can be propagated in nude mice

The natural history of various human breast lesions and cancer in patient is difficult to study since diagnosis is established only after surgical ablation and therefore the clinical course of the undisturbed lesion in vivo can no longer be followed. Development of an experimental system which can propagate and maintain human breast lesions and cancer, similar to those seen in surgical breast specimens, may be a first step in the analyses of these various entities. We report that histomorphologically intact human breast lesions and cancer can be recreated in an experimental system using athymic nude mice. When the dissociated cells from surgical breast cancer specimens are embedded in extracellular matrices (collagen gel, Matrigel, or a mixture of the two) and then transplanted into athymic nude mice, the transplanted cells undergo morphogenesis to reflect their original phenotype. A mixture of the two matrices most closely mirrored the breast tissue in situ when stained with Alcian blue-PAS which stains for mucins. In this mixture, not only is the histomorphology recreated, but well-established clinical molecular markers including the estrogen receptor, c-erbB-2 (HER-2/neu), and aberrant cell proliferation are maintained. With the use of an appropriate extracellular matrices, surgical specimens of human breast lesions and cancer can be analyzed in an in vivo experimental system.

Displacement of carcinomatous epithelium in surgical breast specimens following stereotaxic core biopsy.

A variety of needling procedures in the breast may lead to dislodgment and displacement of fragments of breast carcinoma into tissue outside the target lesion of the needling procedure. To assess how frequently displaced fragments of carcinoma are seen in surgical breast specimens following stereotaxic core needle biopsies, slides were reviewed from 43 consecutive cases of breast carcinoma in which surgical excision and/or mastectomy had been performed following an initial diagnostic stereotaxic 14-gauge core biopsy procedure. In 12 of 43 (28%) cases, displaced carcinomatous fragments were identified outside of the main tumor mass. These patients were subjected to other needling procedures that included local anesthetic injection at the time of core biopsy (43 cases), needle localization (22 of 43 cases), suture placement (18 of 43 cases), and fine-needle aspiration (1 of 43 cases). Attributing carcinomatous displacement solely to the core needle biopsy is complicated by these additional needling procedures. In 18 instances, local anesthetic injection by 25-gauge needle was the only needling procedure other than the core biopsy. In 7 of these 18 (39%) cases, fragments of displaced carcinoma were observed outside the main tumor mass. The authors have previously observed only one case in which a 25-gauge needle was associated with epithelial displacement, suggesting that the core biopsy was more likely to have been the cause of displaced epithelium in these cases. Long-term clinical follow-up will be necessary to determine the biologic and clinical significance of these findings.

Epithelial displacement in surgical breast specimens following needling procedures.

Lesional breast tissue is often subjected to various needling procedures prior to resection and formal histologic examination. In this article, we describe a series of 29 surgical breast specimens in which histologic examination revealed fragments of benign or malignant epithelium displaced in breast stroma or in lymphovascular channels, associated with the traumatic effects of a needling procedure. Follow-up revealed that a variety of different interventions, including needle localization, fine-needle aspiration, infiltration with local anesthetic, core biopsy, and suture placement, preceded the surgical resection in these instances. The displaced epithelial fragments mimic stromal invasion to a variable degree and may represent a potential source of misdiagnosis. The incidence and biological significance of epithelial displacement in this context are as yet unknown.

Human breast cancers respond to growth factors in vivo but not in vitro

Growth response of human breast cancer cells to epidermal growth factor (EGF) and transforming growth factoralpha (TGF-α) was tested both in culture and in vivo in nude mice. Human breast cancer cells were obtained from palpable tumors resulting from xenografted primary breast cancers in nude mice. In collagen gel culture, the breast cancer cells grew autonomously as expanding spherical masses of loosely adherent cells in the basal medium and the supplementation of growth factors had no additional stimulatory effect. To determine whether this in vitro response is reflected in vivo, the collagen gel embedded human breast cancer cells were transplanted into athymic nude mice and the growth response to EGF was studied in vivo. In contrast to the situation in vitro, exogenous EGF was growth promoting in vivo. Our results demonstrate the importance of the combined in vitro-in vivo approach in studying physiologically relevant growth regulation. In addition, the use of collagen gel embedded human breast cancer cells for transplantation studies may more closely model the clinical situation in view of the close histopathological resemblance of the recovered gels to the surgical breast specimens.

A Practical Approach to the Handling of Surgical Breast Specimens

This article discusses the role of the pathologist in the examination of breast surgical specimens and the importance of these observations when clinical decisions on therapeutic approaches for breast cancer are made.

The synthesis of a proteinase inhibitor, alpha-1-antichymotrypsin, by human breast epithelial cells.

The synthesis and release of glycoproteins were studied in organ cultures of human breast surgical specimens and in established breast epithelial cell lines, MCF-7 and MDA-MB-231. Biosynthesis was monitored by the incorporation of 14C-glucosamine. Labeled macromolecules in the culture supernatants were analyzed by biochemical and immunological techniques. One to 8% of the labeled glycoproteins from benign breast and infiltrating ductal carcinoma specimens was precipitated by antibodies produced against human serum alpha-1-antichymotrypsin. Twelve percent of the total glycoproteins from the culture supernatants of the MCF-7 cell line was identified as alpha-1-antichymotrypsin. Both the normal serum and the human breast epithelia-derived proteinase inhibitor can be resolved into similar subclasses by two-dimensional gel electrophoresis. MDA-MD-231 and MCF-7 cells which were extensively washed with EDTA, serum-free medium, and phosphate-buffered saline retain this proteinase inhibitor on their cell surfaces. Three to 4% of the total cell-surface iodinated components was immunoprecipitated by these specific antibodies. Since alpha-1-antichymotrypsin is a potent inhibitor of neutral proteinases such as cathepsin G, the demonstration of its synthesis by benign and malignant breast epithelial cells is of considerable interest. This glycoprotein may represent the epithelia's own protective shield of cell surface components and the cell's attempt to moderate the effects of invading leukocytes. In addition, it may play a regulatory role in the maintenance of three-dimensional glandular structures.

Glycoprotein synthesis as a function of epithelial cell arrangement: Biosynthesis and release of glycoproteins by human breast and prostate cells in organ culture

We demonstrate that a technique is available to investigate glycoprotein synthesis in organ cultures of human breast and prostate surgical specimens where the 3-dimensional epithelial cell arrangement remains intact. Malignant breast and prostate epithelium maintained their capacity to synthesize glycoproteins for at least 3 days as followed by the incorporation of [3H]glucosamine into macromolecules. Over 70% of incorporation was by malignant cells as judged by autoradiography. Labeled glycoproteins were released into glandular lumina and consequently into the culture fluid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed predominantly one group of macrmolecules released with an apparent molecular weight of 48,000 +/- 6,000 daltons. This glycoprotein was found in all of the breast specimens studied, which included 1 medullary, 1 infiltrating lobular, and 8 infiltrating duct carcinomas. The pattern was independent of the availability of estrogen receptors. A similar glycoprotein was also observed in the culture media from a Grade I and a Grade II well-differentiated infiltrating prostate carcinoma. Incorporation was below the level of detection in 4 of 6 cases of benign prostatic hyperplasia. A more complex pattern of labeled glycoproteins was found in the media of a Grade II and a Grade III poorly-differentiated prostate carcinoma. The established human mammary carcinoma cell line MCF-7 synthesized and released a similar 48,000 molecular weight glycoprotein but additional components with larger molecular weights were also released. An intriguing interpretation that 3-dimensional tissue integrity restricts some glycoprotein synthesis is discussed. Cells grown in 2-dimensional monolayers could escape from such a topographic restriction and express additional families of glycoproteins.