RESUMO
A single strand conformation polymorphism analysis on polymerase chain reaction (PCR)-amplified genomic DNA was used for the detection of DNA mutations in acid formalin-fixed, paraffin-embedded tissues. Fifty non-small cell lung cancers were screened for mutations in the exon 5 of the p53 tumor suppressor gene. Structural abnormalities of the gene were found in nine (18%) of the tumors examined. The results show that paraffin-embedded tissues fixed in unbuffered formalin can be a good source of DNA for a mobility shift analysis by neutral polyacrylamide gel electrophoresis. The described procedure allows retrospective genetic studies on paraffin blocks available in all pathology departments.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , DNA/análise , Neoplasias Pulmonares/genética , Mutação/genética , Inclusão em Parafina/métodos , Reação em Cadeia da Polimerase/métodos , Polimorfismo Conformacional de Fita Simples , Sequência de Bases , DNA/efeitos dos fármacos , Fixadores , Formaldeído , Genes p53/genética , Humanos , Dados de Sequência Molecular , Estudos Retrospectivos , Fixação de Tecidos/métodosAssuntos
Neoplasias Mamárias Animais/etiologia , Neoplasias Mamárias Experimentais/patologia , Animais , Feminino , Neoplasias Mamárias Animais/patologia , Neoplasias Mamárias Experimentais/induzido quimicamente , Vírus do Tumor Mamário do Camundongo , Camundongos , Camundongos Endogâmicos/genética , Lesões Pré-Cancerosas/patologiaAssuntos
Neoplasias da Mama/patologia , Mama/citologia , Transformação Celular Neoplásica , Citocinas/biossíntese , Oncogenes , Mama/metabolismo , Neoplasias da Mama/metabolismo , Linhagem Celular , Linhagem Celular Transformada , Citocinas/análise , Citocinas/farmacologia , Células Epiteliais , Epitélio/efeitos dos fármacos , Epitélio/metabolismo , Feminino , Humanos , Interleucina-6/farmacologia , Interleucina-8/farmacologia , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/farmacologiaAssuntos
Neoplasias da Mama/patologia , Mama/patologia , Carcinoma Ductal de Mama/patologia , Receptores de Progesterona/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/metabolismo , Neoplasias da Mama/cirurgia , Carcinoma Ductal de Mama/metabolismo , Carcinoma Ductal de Mama/cirurgia , Células Cultivadas , Epitélio/metabolismo , Epitélio/patologia , Feminino , Fibrose , Humanos , Metástase Linfática , Mastectomia Radical , Pessoa de Meia-Idade , Pós-Menopausa , Pré-Menopausa , Receptores de Estrogênio/análise , Receptores de Estrogênio/biossíntese , Receptores de Progesterona/análiseAssuntos
Biomarcadores Tumorais/análise , Neoplasias da Mama/patologia , Carcinoma Ductal de Mama/patologia , Receptores ErbB/análise , Proteínas Proto-Oncogênicas/análise , Receptores de Estrogênio/análise , Receptores de Progesterona/análise , Ciclo Celular , Divisão Celular , Núcleo Celular/patologia , Feminino , Humanos , Imuno-Histoquímica/métodos , Antígeno Ki-67 , Metástase Linfática , Proteínas de Neoplasias/análise , Estadiamento de Neoplasias , Proteínas Nucleares/análise , Prognóstico , Receptor ErbB-2Assuntos
Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Carcinoma/genética , Carcinoma/patologia , Genes p53 , Mutação , Proteína Supressora de Tumor p53/biossíntese , Sequência de Bases , Neoplasias da Mama/classificação , Carcinoma/classificação , Aberrações Cromossômicas , Primers do DNA , Éxons , Feminino , Humanos , Imuno-Histoquímica , Dados de Sequência Molecular , Oligonucleotídeos Antissenso , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição , Proteína Supressora de Tumor p53/análiseRESUMO
p53 is a tumor suppressor gene, located in the short arm of chromosome 17, which encodes for a nuclear protein involved in the control of cellular growth. Mutations in p53 gene are the most common genetic alterations in a several human cancers, including Non Small Cell Lung Cancer (NSCLC). However, up to now, the role of p53 in the tumour's behaviour and its progression has not been completely clear. We performed immunohistochemical staining for mutated p53 using two monoclonal antibodies, PAb1801 and PAb240, in fresh tumour specimens from 103 consecutive patients who underwent surgery for resectable NSCLC. PAb1801 detects both the normal and mutant form of p53, while PAb240 is specific only for the mutant form and recognizes a denaturation-resistant epitope located between aminoacids 156-335. Both antibodies showed a mainly nuclear staining in neoplastic cells but not in surrounding uninvolved lung tissues. 68 out of 100 (68%) and 37 out of 103 (35.9%) of the cases were positive with PAb1801 and with PAb240, respectively. Tumours from patients with hilar-mediastinal lymph node involvement showed a higher p53 expression, detected by PAb1801, than those without nodal metastases (p = 0.04). Moreover, tumours expressing more than 60% of positive cells with both antibodies showed a significant increase of nodal involvement (p = 0.1; p = 0.03). Furthermore, p53 expression was significantly related to post-surgical stage (p Tumor Stage) (p = 0.04). In addition, we did not find any correlation between p53 expression and proliferating activity evaluated by PCNA, Ki-67 and DNA flow cytometric cell cycle. In conclusion, the evaluation of p53 oncogene expression may identify individuals whose resectable NSCLCs have a more aggressive tumour behaviour.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/química , Neoplasias Pulmonares/química , Proteína Supressora de Tumor p53/análise , Antígenos de Neoplasias/análise , Carcinoma Pulmonar de Células não Pequenas/genética , Núcleo Celular/química , Citometria de Fluxo , Genes p53 , Humanos , Antígeno Ki-67 , Neoplasias Pulmonares/genética , Pessoa de Meia-Idade , Mutação/genética , Proteínas de Neoplasias/análise , Proteínas Nucleares/análise , Ploidias , Antígeno Nuclear de Célula em ProliferaçãoRESUMO
Two small series (nine cases each) of human female breasts were collected to compare the morphological changes of mammary glandular trees contralateral to primary breast cancer and those collateral to symptomatic benign lump. Each whole mammary gland was analysed by a submacroscopic scrutiny method using a stereomicroscope. Interesting and suspicious samples were removed for routine histology. Benign subclinical lesions were indifferently present in both series: Spheric cysts (5:5), sclerosing adenosis (3:3), intraductal papillomas (1:1), fibroadenomas (3:1). On the contrary proliferative epithelial lobular lesions with various degree of atypia i.e. atypical lobules (Grades IV-V according to Wellings), were detected only in the first series (p < 0.01). These data agree completely with the hypothesis of a systemic nature of breast cancer and support indirectly the possible predictive value of atypical lobules in bioptic specimens for the subsequent development of cancer in collateral and/or contralateral breasts.
Assuntos
Neoplasias da Mama/epidemiologia , Mama/patologia , Lesões Pré-Cancerosas/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/patologia , Feminino , Doença da Mama Fibrocística/epidemiologia , Doença da Mama Fibrocística/patologia , Humanos , Pessoa de Meia-Idade , Lesões Pré-Cancerosas/patologia , PrevalênciaRESUMO
A new monoclonal antibody (MoAb), MM 1-80, recognizing a tumor associated epitope of a breast high molecular weight mucin molecule was tested, using the avidin biotin immunoperoxidase method on normal and pathological mammary tissues. The normal mammary ducts and lobules were negative. Fibroadenomas showed a strong intracytoplasmic staining. In apocrine metaplasia, adenosis, and papillomatosis, scattered cells showed intracytoplasmic, luminal border or secretion reactivity. In lobular and ductal hyperplasia the cells showed intracytoplasmic immunoreactivity which, however, became more intense and homogeneous in atypical lesions, i.e. lobular and ductal in-situ carcinomas. The infiltrating carcinomas of different histotype expressed positivity on 98% of the cases (113/115) and axillary metastatic lymph nodes were always positive (20/20). The MoAb was tested on 175 human neoplasias of different origin which were in the majority of the cases negative with the exception of adenocarcinomas of the lung, ovary and bladder. MM1-80 appears to react preferentially with mammary cells undergoing hyperplastic, metaplastic and neoplastic processes. The 1-80 epitope distribution is different in these lesions starting with a predominant luminal expression in benign lesions and becoming strong and cytoplasmic in the malignant breast cell.
Assuntos
Anticorpos Monoclonais , Antígenos de Neoplasias/análise , Antígenos Glicosídicos Associados a Tumores , Neoplasias da Mama/química , Carcinoma/química , Imunoglobulina M , Adulto , Feminino , Humanos , Metástase Linfática , Pessoa de Meia-IdadeRESUMO
Low levels of anti-viral activity, mainly interferon alpha/beta (IFN-alpha/beta), are regularly found in lymphoid tissues of BALB/c mice infected with the C3H strain of mammary tumor virus. At the time of tumor development, significant amounts of anti-viral activity were detected in homogenates of spleen and mammary tumors, but not in blood and normal mammary glands. This activity is pH2-resistant and neutralized by antibody to IFN/alpha-beta. The pathogenetic role of IFN in mammary carcinogenesis was investigated in 2 ways: (a) by treating virus-infected newborn mice with antibody to IFN-alpha/beta, and (b) by giving either the latter antibody or IFN-alpha/beta to virus-free animals transplanted with pre-neoplastic lesions. Mice were treated only for 2 months, starting either 1 week after birth or immediately after tumor transplant. In case (a), treatment with antibody to IFN-alpha/beta shortened the incubation period of mammary carcinomas and decreased the mean survival time. In case (b), anti-IFN antibody did not significantly affect the development of mammary tumors. However, exogenous IFN-alpha/beta markedly reduced both tumor incidence and mortality rate. These results indicate that endogenous IFN-alpha/beta plays a crucial role in the in vivo restriction of the early infectious phase of spontaneous carcinogenesis and that relatively high doses of IFN-alpha/beta may inhibit the progression of pre-neoplastic lesions.
Assuntos
Imunoglobulina G/uso terapêutico , Interferon Tipo I/imunologia , Interferon Tipo I/uso terapêutico , Neoplasias Pulmonares/secundário , Neoplasias Mamárias Experimentais/imunologia , Vírus do Tumor Mamário do Camundongo/imunologia , Envelhecimento/imunologia , Animais , Feminino , Interferon Tipo I/análise , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/terapia , Neoplasias Mamárias Experimentais/microbiologia , Neoplasias Mamárias Experimentais/patologia , Neoplasias Mamárias Experimentais/terapia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Metástase Neoplásica , Baço/imunologiaRESUMO
The growth of a tumour beyond a certain size requires angiogenesis. We assessed whether intensity of angiogenesis correlates with metastasis of non-small-cell lung cancer by counting microvessels and grading their density within the initial carcinomas in 87 T1N0M0 patients. After radical surgery, metastases developed in 22. Both microvessel count and density grades correlated significantly with metastatic disease as well as tumour size and proliferative activity. The likelihood of metastasis increased as the vessel count increased. On multivariate analysis, the microvessel density count was the only independent predictor of metastasis.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/irrigação sanguínea , Neoplasias Pulmonares/irrigação sanguínea , Neovascularização Patológica/patologia , Carcinoma Pulmonar de Células não Pequenas/patologia , Feminino , Humanos , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica/patologiaRESUMO
Spontaneously immortalized human mammary epithelial cells MCF-10A were transfected with an activated c-Ha-ras oncogene. Transfected cells (MCF-10T) acquire a malignant phenotype, as already reported. Studies of 125I-2'-deoxyuridine incorporation in cultures given graded doses of hydrocortisone (HC), cholera toxin (CT), epidermal growth factor (EGF), and transforming growth factor alpha (TGF-alpha) showed that though MCF-10T had become almost independent on exogenous EGF and TGF-alpha, they continued to respond to the synergistic effect of HC and CT plus EGF. Both lines were phenotypically characterized with an immunoradiometric assay in live cells. Expression of MHC class-I molecules, human milk-fat-globule-I antigen, and EGF receptor was reduced in ras-transfected cells, although other differentiation markers were unchanged. Exogenous EGF down-regulated the expression of functional EGF-R, selectively in transformed cells. TGF-alpha failed to modulate EGF-R. In contrast, HC strongly stimulated the expression of EGF-R while depressing MHC class-I molecules. Thus, it appears that in vivo HC may co-operate with TGF-alpha and EGF in promoting the growth of transformed mammary cells. This hormone might also favor the escape from immune surveillance by reducing the expression of surface differentiation markers.
Assuntos
Antígenos de Diferenciação/análise , Linhagem Celular Transformada/efeitos dos fármacos , Fator de Crescimento Epidérmico/farmacologia , Genes ras , Hidrocortisona/farmacologia , Fator de Crescimento Transformador alfa/farmacologia , Mama , Relação Dose-Resposta a Droga , Receptores ErbB/análise , Antígenos HLA/análise , Antígenos de Histocompatibilidade Classe I/análise , Humanos , Proto-Oncogene Mas , TransfecçãoRESUMO
The frequency with which int-1 and int-2 are rearranged in mouse mammary tumors by mouse mammary tumor virus (MMTV)-induced insertional mutagenesis is a consequence of the host genetic background. In 75% of C3H mammary tumors, int-1 is rearranged by MMTV insertion, whereas only 30% of BALB/cfC3H tumors contain a virus-induced rearrangement of int-1. This difference is significant (P less than 0.005) and could not be accounted for by the potentially additive effect of the genetically transmitted Mtv-1-encoded virus in C3H mice. Similarly, MMTV-induced rearrangement of the int-2 gene in mammary tumors of the R111 mouse strain (59%) occurred at a significantly (P less than 0.025) higher frequency than in BALB/cfR111 (25%) mammary tumors. Moreover, in BALB/cfR111 mammary tumors, there is evidence that rearrangement of int-1 and int-2 does not occur independently (P less than 0.025). These results suggest that the long history of inbreeding for high tumor incidence of C3H and R111 mouse strains has selected for the fixation of host mutations which either complement the action of the particular int gene or affect the sensitivity of specific subpopulations of mammary epithelium to infection by particular strains of MMTV.
Assuntos
Rearranjo Gênico , Neoplasias Mamárias Experimentais/microbiologia , Vírus do Tumor Mamário do Camundongo/genética , Animais , Sequência de Bases , Northern Blotting , Southern Blotting , Sondas de DNA , DNA Viral/química , Feminino , Endogamia , Neoplasias Mamárias Experimentais/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Dados de Sequência Molecular , Mutagênese Insercional , Poli A/análise , RNA/análise , RNA Mensageiro , Sequências Repetitivas de Ácido Nucleico , Transcrição GênicaRESUMO
The correlation of the most important prognostic indicators was evaluated in 75 breast cancer cases. Estrogen-progesterone receptors and proliferating activity were analyzed by immunocytochemical methods (ER-ICA, PR-ICA, Ki-67). Both steroid receptors were inversely correlated with the proliferating activity (ER-ICA vs Ki-67, p less than 0.003; PR-ICA vs. Ki-67, p less than 0.0001). No correlation was found between steroid receptors or cell kinetics and tumor size or lymph node status. These findings confirm the relevance of biochemical and kinetic parameters as independent markers in breast cancer and suggest a routine use of the simple immunocytochemical methods in assessing the biological behavior of tumors.
Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/metabolismo , Receptores de Estrogênio/metabolismo , Receptores de Progesterona/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/patologia , Divisão Celular , Feminino , Humanos , Imuno-Histoquímica , Antígeno Ki-67 , Cinética , Pessoa de Meia-Idade , Proteínas Nucleares/metabolismo , PrognósticoRESUMO
The production of monoclonal antibodies against estrogen receptor (ER) and progesterone receptor (PR) has permitted the development of the enzyme immunoassay (EIA) and immunocytochemical assay (ICA) for steroid receptor determination. The results obtained with these two techniques, using the same monoclonal antibodies, were compared in a large series of breast carcinomas (187 for ER and 100 for PR). The correlation between these methods was significant for ER (rs = 0.54) and PR (rs = 0.55) (P less than 0.001) but was lost when the receptor concentrations determined by EIA were less than or equal to 15 and less than or equal to 30 fmol/mg protein for ER and PR, respectively. When these values are considered as cutoffs, the concordance between the two methods was 84.5% for ER and 73% for PR. An analysis of discordant results revealed that low epithelial cellularity generally was present in ICA-positive, EIA-negative specimens, whereas only focal positivity with ICA, or positivity of only normal peripheral mammary ducts and lobules, frequently was found in ICA-negative, EIA-positive tumors. In conclusion, there is good correlation between the results obtained by EIA and ICA methods for detection of ER and PR. The authors suggest that biochemical and histochemical methods for steroid receptors could be considered complementary and used together for the analysis of breast cancer.
Assuntos
Anticorpos Monoclonais , Neoplasias da Mama/metabolismo , Técnicas Imunoenzimáticas , Imuno-Histoquímica/métodos , Receptores de Estrogênio/metabolismo , Receptores de Progesterona/metabolismo , Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
Twenty-eight patients with small cell lung cancer were treated with high dose 4'-epidoxorubicin (EDX). Fifteen patients underwent EDX monotherapy (cumulative dose: 800 mg/m2) while 13 were treated with EDX (cumulative dose: 660 mg/m2) associated with cyclophosphamide, etoposide and cisplatin. A 2D-echo was performed in basal condition, after the third and sixth dose and 2 months after the end of the therapy. In the patients without cardiovascular disease (n = 19) left ventricular end-diastolic volume (EDV), end systolic volume (ESV) and ejection fraction (EF) were unchanged. The patients with coronary artery disease (n = 5) showed a statistically significant decrease in EF with an increase of ESV. All patients with systemic hypertension (n = 4) showed a significant reduction of EF and a significant increase of ESV and EDV.
Assuntos
Epirubicina/efeitos adversos , Coração/efeitos dos fármacos , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biópsia , Carcinoma de Células Pequenas/complicações , Carcinoma de Células Pequenas/tratamento farmacológico , Cisplatino/administração & dosagem , Ciclofosfamida/administração & dosagem , Relação Dose-Resposta a Droga , Ecocardiografia , Epirubicina/administração & dosagem , Etoposídeo/administração & dosagem , Feminino , Humanos , Neoplasias Pulmonares/complicações , Neoplasias Pulmonares/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Miocárdio/patologiaRESUMO
A series of postmenopausal women who had died without noticing any clinical breast disease in their anamnesis (100 cases, age range 46-90 years, average age 62 years) were submitted to bilateral subcutaneous mastectomy during autopsy in order to evaluate the morphologic profile of asymptomatic mammary glands, at different ages. Submacroscopic changes were found and removed to be processed for histology. Results were as follows: a) 46% of cases did not show any change; b) 54% of cases showed benign changes, namely a fibrocystic condition; c) 14% of cases had in addition epithelial lobular hyperplasia with low grade atypia and d) 3% of cases showed atypical borderline lobules (ABL), i.e., terminal ductal-lobular units characterized by severe epithelial atypia. Such lesions cannot be easily distinguished from "in situ" carcinoma, and are currently considered at morphologic risk for subsequent cancer when found in breast biopsies. Our data show that: 1) ABL do not represent a common finding in women who never complained of breast pathology during life; 2) ABL are not related to older age; 3) Fibrocystic condition is quite frequent at subclinical levels also in asymptomatic aging women. The latter statement confirms the opinion that fibrocystic condition should be considered as a common "functional" change. On the contrary, the rarity of ABL gives us a further indirect evidence of their possible precancerous significance. The risk of subsequent development of cancer from the collateral mammary gland could be theoretically higher when ABL are found in breast biopsies of fertile and premenopausal women, who have a longer period of life expectation.
Assuntos
Mama/patologia , Doença da Mama Fibrocística/patologia , Menopausa , Lesões Pré-Cancerosas/patologia , Idoso , Idoso de 80 Anos ou mais , Autopsia , Feminino , Humanos , Pessoa de Meia-Idade , Fatores de RiscoRESUMO
Here we report that the slow-transforming helper component of Friend murine leukaemia virus (Fr-MLV), which produces lymphoid leukaemias in normal mice, induces erythroleukaemia in mice given syngeneic pituitary grafts (SPG). Newborn mice were infected with Fr-MLV and, at one month of age, were transplanted with two pituitary glands under the kidney capsule. Sham-operated infected mice and uninfected transplanted mice served as controls. SPG selectively reduced the mean survival times of infected mice. Histopathology showed that, while most infected non-transplanted mice developed lymphoid leukaemias, virtually all Fr-MLF-infected mice given SPG developed erythroleukaemias. Experiments in vitro showed that Fr-MLV infection markedly depressed concanavalin A induced DNA synthesis in cells from spleen, thymus and lymph nodes. Addition of prolactin or growth hormone further suppressed lectin-induced mitogenesis of lymphoid cells from infected mice, but failed to influence the response of uninfected controls. These experiments indicate that, in mice, pituitary hormones modulate the development and the histological features of Fr-MLV induced leukaemias, and suggest that endocrine-immunological interactions play a role in retrovirus induced tumorigenesis.
Assuntos
Leucemia Eritroblástica Aguda/etiologia , Hipófise/transplante , Animais , Feminino , Vírus da Leucemia Murina de Friend , Hormônio do Crescimento/farmacologia , Leucemia Eritroblástica Aguda/mortalidade , Leucemia Eritroblástica Aguda/patologia , Leucemia Linfoide/etiologia , Camundongos , Camundongos Endogâmicos BALB C , Prolactina/farmacologiaRESUMO
Eleven cases of thyroid lymphoma were studied by the immunoperoxidase avidin-biotin technique with calcitonin and somatostatin rabbit antisera. In 6 cases of non-Hodgkin lymphoma, in thyroid tissue residual to the lymphomatous infiltration, the C cell density was markedly increased and clustering was often observed; the C cells often took part in the follicular lining, frequently with polar distribution; these elements displayed a strong positivity for calcitonin, while the number of somatostatin-containing cells was lower and the staining less intense. In the only case of Hodgkin's lymphoma of the thyroid gland the staining was negative; in other 4 cases of non-Hodgkin lymphoma no residual thyroid tissue was found and the staining was also negative. As Hashimoto's thyroiditis is often associated with thyroid lymphoma, 13 cases of Hashimoto's thyroiditis were also studied; no C cells were observed and both stainings were negative. These data show that an increase in the C cell number may be a hallmark of thyroid lymphoma and that hyperplastic C cells show an intensive positivity for calcitonin. On the other hand, C cell hyperplasia is not present in Hashimoto's thyroiditis, in spite of the close association with thyroid lymphoma. Furthermore, we provide evidence that somatostatin-containing cells are present both in normal thyroid glands and in thyroid lymphoma.
Assuntos
Calcitonina/análise , Linfoma/análise , Somatostatina/análise , Glândula Tireoide/análise , Neoplasias da Glândula Tireoide/análise , Tireoidite Autoimune/patologia , Doença de Hodgkin/análise , Doença de Hodgkin/patologia , Humanos , Hiperplasia , Imuno-Histoquímica , Linfoma/patologia , Linfoma não Hodgkin/análise , Linfoma não Hodgkin/patologia , Glândula Tireoide/citologia , Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/patologiaRESUMO
ER-D5 is a recently identified protein related to estrogen receptors (ER). Generally ER measurement requires fresh frozen tissue and for ER-D5 assay ethanol (E) fixation of the specimen is recommended. We evaluated the possibility of immunocytochemical detection of ER-D5 in routine formalin-fixed (F) sections in 51 breast cancers comparing the results with those obtained in the same specimens using E as fixative. The results of ER-D5 assay were expressed by the staining index (SI) taking values greater than or equal to 5 as positive. In all tumors ER was also assayed by a biochemical method (DCCA). The sensitivity of ER-D5 detection in F was only 33.3%, while the specificity was 94.4%. A lower cut-off value of SI for F sections (greater than or equal to 2) increased the sensitivity to 66.6%, leaving the specificity unchanged. A strong correlation was found between the SI of ER-D5 in E and F (p less than 0.001). The SI of ER-D5 in F sections was also well correlated with ER concentrations (p less than 0.001). These results suggest that immunocytochemical determination of ER-D5 in routine sections may be useful in retrospective studies of hormone dependence in breast cancer.