RESUMEN
To identify markers of non-response to neoadjuvant chemotherapy (NAC) that could be used in the adjuvant setting. Sixteen pathologists of the European Working Group for Breast Screening Pathology reviewed the core biopsies of breast cancers treated with NAC and recorded the clinico-pathological findings (histological type and grade; estrogen, progesterone receptors, and HER2 status; Ki67; mitotic count; tumor-infiltrating lymphocytes; necrosis) and data regarding the pathological response in corresponding surgical resection specimens. Analyses were carried out in a cohort of 490 cases by comparing the groups of patients showing pathological complete response (pCR) and partial response (pPR) with the group of non-responders (pathological non-response: pNR). Among other parameters, the lobular histotype and the absence of inflammation were significantly more common in pNR (p < 0.001). By ROC curve analyses, cut-off values of 9 mitosis/2 mm(2) and 18% of Ki67-positive cells best discriminated the pNR and pCR + pPR categories (p = 0.018 and < 0.001, respectively). By multivariable analysis, only the cut-off value of 9 mitosis discriminated the different response categories (p = 0.036) in the entire cohort. In the Luminal B/HER2- subgroup, a mitotic count <9, although not statistically significant, showed an OR of 2.7 of pNR. A lobular histotype and the absence of inflammation were independent predictors of pNR (p = 0.024 and <0.001, respectively). Classical morphological parameters, such as lobular histotype and inflammation, confirmed their predictive value in response to NAC, particularly in the Luminal B/HER2- subgroup, which is a challenging breast cancer subtype from a therapeutic point of view. Mitotic count could represent an additional marker but has a poor positive predictive value.
Asunto(s)
Neoplasias de la Mama/tratamiento farmacológico , Mitosis/genética , Terapia Neoadyuvante , Biomarcadores de Tumor/genética , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Proliferación Celular/genética , Supervivencia sin Enfermedad , Resistencia a Antineoplásicos , Estrógenos/genética , Femenino , Humanos , Receptor ErbB-2/genética , Receptores de Progesterona/genéticaRESUMEN
Microinvasion is the smallest morphologically identifiable stage of invasion. Its presence and distinction from in situ carcinoma may have therapeutic implications, and clinical staging also requires the recognition of this phenomenon. Microinvasion is established on the basis of several morphological criteria, which may be difficult and not perfectly reproducible among pathologists. The aim of this study was to assess the consistency of diagnosing microinvasion in the breast on traditional haematoxylin and eosin (HE) stained slides and to evaluate whether immunohistochemistry (IHC) for myoepithelial markers could improve this. Digital images were generated from representative areas of 50 cases stained with HE and IHC for myoepithelial markers. Cases were specifically selected from the spectrum of in situ to microinvasive cancers. Twenty-eight dedicated breast pathologists assessed these cases at different magnifications through a web-based platform in two rounds: first HE only and after a washout period by both HE and IHC. Consistency in the recognition of microinvasion significantly improved with the use of IHC. Concordance rates increased from 0.85 to 0.96, kappa from 0.5 to 0.85, the number of cases with 100% agreement rose from 9/50 to 25/50 with IHC and the certainty of diagnosis also increased. The use of IHC markedly improves the consistency of identifying microinvasion. This corroborates previous recommendations to use IHC for myoepithelial markers to clarify cases where uncertainty exists about the presence of microinvasion. Microinvasive carcinoma is a rare entity, and seeking a second opinion may avoid overdiagnosis.
Asunto(s)
Biomarcadores de Tumor/análisis , Neoplasias de la Mama/patología , Carcinoma/patología , Inmunohistoquímica/métodos , Metástasis de la Neoplasia/diagnóstico , Femenino , Humanos , Variaciones Dependientes del Observador , Patología Clínica/métodos , Patología Clínica/normasRESUMEN
It is now widely recognised that classifying ductal carcinoma in situ (DCIS) of the breast and diagnosing atypical ductal hyperplasia are associated with significant interobserver variation. Two possible reasons for this inconsistency are differences in the interpretation of specified histological features and field selection where morphology is heterogeneous. In order to investigate the relative contribution of these two factors to inconsistent interpretation of intraductal proliferations, histological sections of 32 lesions were sent to 23 European pathologists followed 3 years later by images of small parts of these sections. Kappa statistics for diagnosing hyperplasia of usual type, atypical ductal hyperplasia and ductal carcinoma in situ were 0.54, 0.35 and 0.78 for sections and 0.47, 0.29 and 0.78 for images, respectively, showing that most of the inconsistency is due to differences in morphological interpretation. Improvements can thus be expected only if diagnostic criteria or methodology are changed. In contrast, kappa for classifying DCIS by growth pattern was very low at 0.23 for sections and better at 0.47 for images, reflecting the widely recognised variation in the growth pattern of DCIS. Higher kappa statistics were obtained when any mention of an individual growth pattern was included in that category, thus allowing multiple categories per case; but kappa was still higher for images than sections. Classifying DCIS by nuclear grade gave kappa values of 0.36 for sections and 0.49 for images, indicating that intralesional heterogeneity has hitherto been underestimated as a cause of inconsistency in classifying DCIS by this method. More rigorous assessment of the proportions of the different nuclear grades present could lead to an improvement in consistency.
Asunto(s)
Neoplasias de la Mama/diagnóstico , Mama/patología , Carcinoma in Situ/diagnóstico , Carcinoma Ductal de Mama/diagnóstico , Neoplasias de la Mama/clasificación , Carcinoma in Situ/clasificación , Carcinoma Ductal de Mama/clasificación , Femenino , Humanos , Hiperplasia/diagnóstico , Variaciones Dependientes del ObservadorRESUMEN
Controversies and inconsistencies regarding the pathological work-up of sentinel lymph nodes (SNs) led the European Working Group for Breast Screening Pathology (EWGBSP) to review published data and current evidence that can promote the formulation of European guidelines for the pathological work-up of SNs. After an evaluation of the accuracy of SN biopsy as a staging procedure, the yields of different sectioning methods and the immunohistochemical detection of metastatic cells are reviewed. Currently published data do not allow the significance of micrometastases or isolated tumour cells to be established, but it is suggested that approximately 18% of the cases may be associated with further nodal (non-SN) metastases, i.e. approximately 2% of all patients initially staged by SN biopsy. The methods for the intraoperative and molecular assessment of SNs are also surveyed.
Asunto(s)
Neoplasias de la Mama/patología , Biopsia del Ganglio Linfático Centinela/métodos , Femenino , Humanos , Metástasis de la Neoplasia/patología , Variaciones Dependientes del Observador , Guías de Práctica Clínica como Asunto , Valor Predictivo de las Pruebas , Pronóstico , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Sensibilidad y Especificidad , Biopsia del Ganglio Linfático Centinela/normasRESUMEN
The increased detection of ductal carcinoma in situ (DCIS) by mammographic screening, the greater use of breast-conserving surgery, and the recognition that certain histological subtypes are associated with a greater risk of local recurrence has led to the formulation of several new classifications of DCIS in recent years. There are, however, no data concerning the degree of consistency with which these schemes can be applied by reasonable numbers of pathologists. Thirty-three cases of DCIS were thus examined by a working group of 23 European pathologists who categorized them using five recently published classifications: (1) that of the European Pathologists' Working Group based on differentiation (a combination of nuclear grade and cell polarization) with categories of poorly, intermediately, and well differentiated; (2) one based entirely on nuclear grade with categories of high, intermediate, and low, currently in use in the UK national and EC-funded breast screening programs; (3) the same classification in which only two categories, high nuclear grade and other, were used; (4) the Van Nuys system in which lesions are divided into high grade, non-high grade with necrosis and non-high grade without necrosis; and (5) a two-category classification based entirely on the presence or absence of comedo necrosis. Of the three systems with three categories, Van Nuys gave the highest overall kappa statistic of 0.42. Others gave similar values of 0.37 and 0.35 showing that assessing cell polarization in addition to nuclear grade neither improves nor worsens consistency. In all three systems, the middle category was associated with the lowest value for kappa. Of the two systems with two categories, that based on nuclear grade gave the highest overall kappa of 0.46 and that based on comedo necrosis the lowest of 0.34. The most robust histological features were thus high- and low-grade nuclei and necrosis as long as the latter did not involve the recognition of a comedo growth pattern. These values probably represent the maximum achievable, at least by reasonable numbers of pathologists in everyday practice. They are better than those previously reported for classification based entirely on architecture, but further improvement is needed.
Asunto(s)
Neoplasias de la Mama/clasificación , Carcinoma in Situ/clasificación , Carcinoma Ductal de Mama/clasificación , Neoplasias de la Mama/patología , Carcinoma in Situ/patología , Carcinoma Ductal de Mama/patología , Europa (Continente) , Femenino , Humanos , Variaciones Dependientes del ObservadorRESUMEN
AIMS: Cyclo-oxygenases 1 and 2 (COX-1 and COX-2) are key enzymes in prostaglandin biosynthesis. COX-2 is induced by a wide variety of stimuli, and present during inflammation. COX-2 overexpression has been observed in colon, head and neck, lung, prostate, stomach, and breast cancer. In colon and gastric cancer, COX-2 expression was associated with angiogenesis. The aim of this study was to determine the relation between COX-2 expression and angiogenesis in breast cancer, and to correlate the expression of this enzyme with classic clinicopathological parameters. METHODS: COX-2 expression was investigated by immunohistochemistry and western blotting analysis. The expression of COX-2 was then related to age, histological grade, nodal status, oestrogen receptor status, p53 expression,c-erb-B2 overexpression, mitotic counts, MIB-1 labelling index, apoptotic index, sialyl-Tn expression, transforming growth factor alpha expression, microvessel density, and disease free survival in 46 patients with invasive ductal breast carcinoma. RESULTS: By means of immunohistochemistry, COX-2 expression was detected in eight of the 46 carcinomas studied. Western blotting showed COX-2 protein expression in the same breast tumours, but not in normal adjacent tissues. The density of microvessels immunostained with anti-F-VIII related antigen was significantly higher in patients with COX-2 expression than in those without expression (p = 0.03). In addition, COX-2 was significantly associated with the presence of sialyl-Tn expression (p = 0.02), lymph node metastasis (p = 0.03), a high apoptotic index (p = 0.03), and a short disease free survival (p = 0.03) in univariate analyses. CONCLUSIONS: These data suggest that COX-2 expression is associated with angiogenesis, lymph node metastasis, and apoptosis in human breast cancer. Moreover, these results warrant further studies with larger series of patients to confirm the association with short disease free survival in patients with breast cancer.
Asunto(s)
Neoplasias de la Mama/irrigación sanguínea , Carcinoma Ductal de Mama/irrigación sanguínea , Isoenzimas/metabolismo , Proteínas de Neoplasias/metabolismo , Neovascularización Patológica/enzimología , Prostaglandina-Endoperóxido Sintasas/metabolismo , Adulto , Anciano , Apoptosis , Western Blotting , Neoplasias de la Mama/enzimología , Neoplasias de la Mama/patología , Carcinoma Ductal de Mama/enzimología , Carcinoma Ductal de Mama/secundario , Ciclooxigenasa 2 , Supervivencia sin Enfermedad , Femenino , Humanos , Técnicas para Inmunoenzimas , Metástasis Linfática , Proteínas de la Membrana , Persona de Mediana Edad , Índice MitóticoRESUMEN
AIMS: One of the limitations of fine needle aspiration biopsy (FNAB) of the breast is in distinguishing invasive carcinoma (IDC) from ductal carcinoma in situ (DCIS). It has been proposed that the presence of myoepithelial cells overlying epithelial malignant cell clusters suggests DCIS. However, the recognition of myoepithelial cells in aspirates may be difficult. The aim of this study was to investigate a new nuclear myoepithelial cell marker, p63, a p53 homologue nuclear transcription factor, in a series of breast FNABs in an attempt to distinguish IDC from DCIS. METHODS: Papanicolaou stained smears from eight cases of pure DCIS and 15 cases of pure IDC with a histologically confirmed diagnosis were submitted to immunocytochemical analysis using the antibody 4A4 against p63. Two pathologists evaluated the presence of p63 positive cells overlying malignant cell clusters and admixed with malignant cells. The frequency of p63 positive cells in DCIS and IDC was compared using Fisher's exact test. RESULTS: p63 consistently stained the nuclei of myoepithelial cells, either overlying malignant cell clusters and/or admixed with malignant cells. p63 positive myoepithelial cells were seen in all DCIS cases and in nine of the 15 cases of IDC (p = 0.0375). In eight cases (three DCIS and five IDC), scattered p63+ epithelial malignant cells were seen. CONCLUSIONS: Although p63 positive myoepithelial cells are found more frequently in DCIS cases, their presence cannot be used as a criterion to rule out invasion in breast FNABs because they are present in up to 60% of invasive cases.
Asunto(s)
Biomarcadores de Tumor/análisis , Neoplasias de la Mama/patología , Carcinoma Ductal de Mama/patología , Carcinoma Intraductal no Infiltrante/patología , Proteínas de la Membrana , Fosfoproteínas/análisis , Transactivadores/análisis , Adulto , Anciano , Anciano de 80 o más Años , Biopsia con Aguja , Neoplasias de la Mama/química , Carcinoma Ductal de Mama/química , Carcinoma Intraductal no Infiltrante/química , Proteínas de Unión al ADN , Diagnóstico Diferencial , Femenino , Genes Supresores de Tumor , Humanos , Técnicas para Inmunoenzimas , Persona de Mediana Edad , Estudios Retrospectivos , Factores de Transcripción , Proteínas Supresoras de TumorRESUMEN
AIM: To study the immunoexpression and mutational status of c-KIT and PDGFRA in a series of benign and malignant phyllodes tumours of the breast. MATERIAL/METHODS: Nineteen phyllodes tumours (13 benign and six malignant) were analysed by immunohistochemistry for the expression of c-KIT and PDGFRA. Direct sequencing of exons 9, 11, 13, and 17 of the c-KIT gene and exons 12 and 18 of PDGFRA was performed to check the mutational status of these two genes. RESULTS: c-KIT expression was found in 12 of the 19 cases (six of the 13 benign cases and all six malignant ones) and PDGFRA expression was seen in two of the 19 cases (one benign and one malignant case); the 2415 C>T alteration in exon 17 of the c-KIT gene was found in two cases (both benign); the intronic insertion IVS17-50insT and the 2866 G>T alteration in the coding region of exon 18 of the PDGFRA gene were also found in two cases (one malignant and one benign). However, the activating mutations described for these genes in gastrointestinal stromal tumours were not present. CONCLUSION: c-KIT expression is a frequent finding in phyllodes tumours, particularly in malignant cases; however, no activating mutations similar to those described for gastrointestinal stromal tumours were found. The PDGFRA does not seem to be an alternative pathway to tumour development in phyllodes tumours because neither expression nor activating mutations were noteworthy.
Asunto(s)
Biomarcadores de Tumor/análisis , Neoplasias de la Mama/química , Tumor Filoide/química , Proteínas Proto-Oncogénicas c-kit/análisis , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/análisis , Biomarcadores de Tumor/genética , Neoplasias de la Mama/metabolismo , Análisis Mutacional de ADN , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Inmunohistoquímica/métodos , Tumor Filoide/metabolismo , Proteínas Proto-Oncogénicas c-kit/genética , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/genéticaRESUMEN
A detailed analysis of the consistency with which pathologists from 12 different European countries diagnose and classify breast disease was undertaken as part of the quality assurance programme of the European Breast Screening Pilot Network funded by the Europe against Cancer Programme. Altogether 107 cases were examined by 23 pathologists in 4 rounds. Kappa statistics for major diagnostic categories were: benign (not otherwise specified) 0.74, atypical ductal hyperplasia (ADH) 0.27, ductal carcinoma in situ (DCIS) 0.87 and invasive carcinoma 0.94. ADH was the majority diagnosis in only 2 cases but was diagnosed by at least 2 participants in another 14, in 9 of which the majority diagnosis was benign (explaining the relatively low kappa for this category). DCIS in 4 (all low nuclear grade) and invasive carcinoma (a solitary 1-mm focus) in 1. The histological features of these cases were extremely variable; although one feature that nearly all shared was the presence of cells with small, uniform, hyperchromatic nuclei and a high nucleo-cytoplasmic ratio. The majority diagnosis was DCIS in 33 cases; kappa for classifying by nuclear grade was 0.38 using three categories and 0.46 when only two (high and other) were used. When ADH was included with low nuclear grade DCIS there was only a slight improvement in kappa. Size measurement of DCIS was less consistent than that of invasive carcinoma. The majority diagnosis was invasive carcinoma in 57 cases, the size of the majority being 100% in 49. The remainder were either special subtypes (adenoid cystic, tubular, colloid, secretory, ductal/medullary) or possible microinvasive carcinomas. Subtyping was most consistent for mucinous (kappa, 0.92) and least consistent for medullary carcinomas (kappa, 0.56). Consistency of grading using the Nottingham method was moderate (kappa=0.53) and consistency of diagnosing vascular invasion, fair (kappa=0.38). There was no tendency for consistency to improve from one round to the next, suggesting that further improvements are unlikely without changes in guidelines or methodology.
Asunto(s)
Enfermedades de la Mama/diagnóstico , Neoplasias de la Mama/diagnóstico , Enfermedades de la Mama/patología , Neoplasias de la Mama/patología , Carcinoma/diagnóstico , Femenino , Guías como Asunto , Humanos , Hiperplasia , Invasividad Neoplásica , PronósticoRESUMEN
To assess the variability of oestrogen receptor (ER) testing using immunocytochemistry, centrally stained and unstained slides from breast cancers were circulated to the members of the European Working Group for Breast Screening Pathology, who were asked to report on both slides. The results showed that there was almost complete concordance among readers (kappa=0.95) in ER-negative tumours on the stained slide and excellent concordance among readers (kappa=0.82) on the slides stained in each individual laboratory. Tumours showing strong positivity were reasonably well assessed (kappa=0.57 and 0.4, respectively), but there was less concordance in tumours with moderate and low levels of ER, especially when these were heterogeneous in their staining. Because of the variation, the Working Group recommends that laboratories performing these stains should take part in a external quality assurance scheme for immunocytochemistry, should include a tumour with low ER levels as a weak positive control and should audit the percentage positive tumours in their laboratory against the accepted norms annually. The Quick score method of receptor assessment may also have too many categories for good concordance, and grouping of these into fewer categories may remove some of the variation among laboratories.
Asunto(s)
Neoplasias de la Mama/metabolismo , Inmunohistoquímica/normas , Receptores de Estrógenos/metabolismo , Coloración y Etiquetado/normas , Unión Europea , Femenino , Humanos , Control de Calidad , Reproducibilidad de los ResultadosRESUMEN
AIMS: To evaluate aspects of the current practice of sentinel lymph node (SLN) pathology in breast cancer via a questionnaire based survey, to recognise major issues that the European guidelines for mammography screening should address in the next revision. METHODS: A questionnaire was circulated by mail or electronically by the authors in their respective countries. Replies from pathology units dealing with SLN specimens were evaluated further. RESULTS: Of the 382 respondents, 240 European pathology units were dealing with SLN specimens. Sixty per cent of these units carried out intraoperative assessment, most commonly consisting of frozen sections. Most units slice larger SLNs into pieces and only 12% assess these slices on a single haematoxylin and eosin (HE) stained slide. Seventy one per cent of the units routinely use immunohistochemistry in all cases negative by HE. The terms micrometastasis, submicrometastasis, and isolated tumour cells (ITCs) are used in 93%, 22%, and 71% of units, respectively, but have a rather heterogeneous interpretation. Molecular SLN staging was reported by only 10 units (4%). Most institutions have their own guidelines for SLN processing, but some countries also have well recognised national guidelines. CONCLUSIONS: Pathological examination of SLNs throughout Europe varies considerably and is not standardised. The European guidelines should focus on standardising examination. They should recommend techniques that identify metastases > 2 mm as a minimum standard. Uniform reporting of additional findings may also be important, because micrometastases and ITCs may in the future be shown to have clinical relevance.
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Neoplasias de la Mama/patología , Práctica Profesional/estadística & datos numéricos , Biopsia del Ganglio Linfático Centinela/normas , Biomarcadores de Tumor/análisis , Femenino , Encuestas de Atención de la Salud , Humanos , Inmunohistoquímica , Cuidados Intraoperatorios/métodos , Cuidados Intraoperatorios/normas , Metástasis Linfática , Guías de Práctica Clínica como Asunto , Biopsia del Ganglio Linfático Centinela/métodos , Encuestas y CuestionariosRESUMEN
We describe a method of immunocytochemically assessing estrogen receptor (ER) status on alcohol-fixed smears obtained by fine-needle aspiration (FNA) from breast cancer patients, using a commercially available monoclonal antibody (1D5) with microwave oven processing. A series of 31 cases of aspirates from breast cancer were analysed and the results were compared with assessment by ER immunocytochemical assay using the same procedure on formalin-fixed tissue and with assessment by ER-ICA assay on frozen sections. The results were scored semiquantitatively using a five grade scoring system. Of the 31 cases examined, 21 were positive at least by two methods and 10 were negative for all three determinations. The results obtained in the ER immunocytochemical assay on aspirates and paraffin-sections using the antibody 1D5 and those obtained on frozen sections using the antibody H222 were closely similar. In only one case was it not possible to interpret the reaction in the cytological specimen because there was a strong background in the smear. In general, we obtained more intense positivity with the antibody 1D5 in aspirates and formalin-fixed material than with the antibody H222 in frozen sections. The scoring results of the three methods were almost identical. We conclude that the application of ER method on alcohol-fixed smears will eliminate the need for using a special fixation procedure and will provide several advantages, such as: improvement in morphological concomitant analysis, utilization whenever malignancy is found without necessity to re-aspirate the patient, and adequacy of archival material.
Asunto(s)
Anticuerpos Monoclonales , Biopsia con Aguja , Neoplasias de la Mama/química , Inmunohistoquímica/métodos , Microondas , Receptores de Estrógenos/análisis , Adulto , Anciano , Femenino , Secciones por Congelación , Humanos , Persona de Mediana Edad , Parafina , Sensibilidad y Especificidad , Adhesión del TejidoRESUMEN
Malignant transformation is frequently associated with abnormal expression of cell surface carbohydrates. Sialyl-Tn (STn) is a core carbohydrate antigen of tumor-associated mucin formed by the premature 2-6 sialylation of N-acetylgalactosamine. In an attempt to verify whether this antigen is restricted to malignant cells, we studied 30 cases of fine-needle aspiration (FNA) cytology from mammographically detected breast lesions. The rationale for choosing this material was the acknowledged difficulty in diagnosing cytologically small breast lesions, especially epithelial intraductal proliferations. The cases were divided in benign lesions (two fibroadenomas and ten ductal hyperplasias) and malignant lesions (16 ductal carcinomas). Ten of sixteen malignant cases (62.5%) were positive for STn. Five of fourteen benign cases (35.7%) were also positive for STn (two fibroadenomas and three ductal hyperplasias). The most consistent positive results in benign lesions resulted from cases that displayed apocrine metaplasia, although positivity has also been observed in ductal cells without metaplasia. We did not find statistical significant differences among STn expression in benign and malignant breast lesions detected by FNA (P = 0.14). Thus, we conclude that STn is neither specific nor sensitive for detection of malignancy in FNA from mammographically detected breast lesions.
Asunto(s)
Antígenos de Carbohidratos Asociados a Tumores/análisis , Biomarcadores de Tumor/análisis , Neoplasias de la Mama/diagnóstico , Adulto , Anciano , Biopsia con Aguja , Neoplasias de la Mama/diagnóstico por imagen , Neoplasias de la Mama/patología , Femenino , Humanos , Mamografía , Persona de Mediana EdadRESUMEN
OBJECTIVE: To verify the potential role of fine needle aspiration (FNA) cytology in obtaining malignant cells from primary breast cancer for establishment of a primary breast cancer cell line. STUDY DESIGN: In four patients with primary breast cancer subjected to FNA for diagnostic purposes, we attempted to establish primary cultures. We successfully obtained one primary cell line, originating in micropapillary invasive breast carcinoma. FNA material obtained under sterile conditions was centrifuged, and the cell pellet was washed with Dulbecco Modified Medium. The resulting suspension was seeded in 25-cm2 tissue culture flasks. The flasks were maintained with released caps in a 37 degrees C incubator with a humidified atmosphere of 5% CO2 in air. After one week, cells attached to the bottom of the flasks and began proliferating. When a culture became confluent, the cells were treated with 0.05% trypsin/0.02% EDTA in a PBS solution and subcultured. The flasks were observed daily with an inverted microscope, and culture passages were performed weekly. RESULTS: The cell line obtained was named I2FPRW and exhibited morphologic and immunohistochemical features of epithelial cells of mammary origin. The cells were positive for cytokeratins (AE1/AE3 and CK 7), EMA and c-erbB-2. At this writing, this cell line was in the 15th passage of subculturing in the flasks with 10% FBS. CONCLUSION: In the present study we demonstrated that is possible to establish a breast cancer cell line from material obtained by FNA cytology. FNA seems to be a valuable method of obtaining malignant cells from breast cancer able to grow free of fibroblasts in cell cultures.
Asunto(s)
Biopsia con Aguja , Neoplasias de la Mama/patología , Adulto , Animales , Células Epiteliales/patología , Femenino , Humanos , Inmunohistoquímica , Ratones , Ratones Desnudos , Invasividad Neoplásica , Trasplante de Neoplasias , Células Tumorales CultivadasRESUMEN
Germline mutations in the BRCA1 gene confer an increased susceptibility to breast and ovarian cancer. Approximately 460 distinct mutations were founded scattered throughout the whole gene. However, several mutations were detected repeatedly in individuals of the same ethnic origin. There are no systematic studies concerning mutations in BRCA1 gene in Portugal. The aim of this study is to identify mutations inh this gene in patients with breast and/or ovarian cancer of Portuguese origin. Thirty-three cases of breast cancer and 3 of ovarian cancer were selected according to early age of onset and family history. Mutation screening of this gene was done by Protein Truncation Test (PTT) and Fragment analysis. Two de novo mutations were identified: 1) A frameshift mutation localised in exon 11 of BRCA1, was identify by the two techniques in an ovarian cancer case. Direct sequential revealed a deletion of 4 nucleotides (3444delAAAT); 2) An alteration in intron 18 (IVS 18 + 80delT) was observed in a breast cancer case. This is probably the first description of mutations in a series of patients with breast and/or cancer in the Portuguese population. The mutations identified in this study have not been previously described in other populations according to the Breast Cancer Information Core web site.
Asunto(s)
Neoplasias de la Mama/genética , Genes BRCA1/genética , Neoplasias Ováricas/genética , Eliminación de Secuencia/genética , Femenino , HumanosRESUMEN
We reviewed 25 patients submitted to transthoracic fine-needle aspiration (TFNA) who had a final diagnosis of pulmonary tuberculosis. In all cases, bacteriological stains and cultures of sputum and bronchial washing had been performed before admission and were negative. According to the material obtained from the procedure, the aspirates were divided in three groups: diagnostic (Ziehl-Neelsen and/or culture positive, n = 8, 32%), suggestive (granulomatous inflammatory changes, n = 10, 40%) and inconclusive (nonspecific inflammatory changes, isolated giant cells and/or blood, n = 7, 28%). On chest X-ray, 12 patients had opacities with the greatest diameter not exceeding 4 cm. All aspirates in the diagnostic group were from patients with this type of lesion, while all the inconclusive aspirates belonged to patients with larger lesions. As complications, 1 patient needed thoracic drainage for pneumothorax and 3 patients had haemoptyses. Thus TFNA has a place in the diagnosis of suspected pulmonary tuberculosis when more simple methods have failed, and its effectiveness seems to be increased when the lesions do not exceed 4 cm in diameter.