RESUMO
BACKGROUND: Mutations in the androgen receptor (AR) ligand-binding domain (LBD), such as F877L and T878A, have been associated with resistance to next-generation AR-directed therapies. ARN-509-001 was a phase I/II study that evaluated apalutamide activity in castration-resistant prostate cancer (CRPC). Here, we evaluated the type and frequency of 11 relevant AR-LBD mutations in apalutamide-treated CRPC patients. PATIENTS AND METHODS: Blood samples from men with nonmetastatic CRPC (nmCRPC) and metastatic CRPC (mCRPC) pre- or post-abiraterone acetate and prednisone (AAP) treatment (≥6 months' exposure) were evaluated at baseline and disease progression in trial ARN-509-001. Mutations were detected in circulating tumor DNA using a digital polymerase chain reaction-based method known as BEAMing (beads, emulsification, amplification and magnetics) (Sysmex Inostics' GmbH). RESULTS: Of the 97 total patients, 51 had nmCRPC, 25 had AAP-naïve mCRPC, and 21 had post-AAP mCRPC. Ninety-three were assessable for the mutation analysis at baseline and 82 of the 93 at progression. The overall frequency of detected AR mutations at baseline was 7/93 (7.5%) and at progression was 6/82 (7.3%). Three of the 82 (3.7%) mCRPC patients (2 AAP-naïve and 1 post-AAP) acquired AR F877L during apalutamide treatment. At baseline, 3 of the 93 (3.2%) post-AAP patients had detectable AR T878A, which was lost after apalutamide treatment in 1 patient who continued apalutamide treatment for 12 months. CONCLUSIONS: The overall frequency of detected mutations at baseline (7.5%) and progression (7.3%) using the sensitive BEAMing assay was low, suggesting that, based on this assay, AR-LBD mutations such as F877L and T878A are not common contributors to de novo or acquired resistance to apalutamide. CLINICALTRIALS.GOV IDENTIFIER: NCT01171898.
Assuntos
Antagonistas de Androgênios/uso terapêutico , Mutação Puntual , Neoplasias de Próstata Resistentes à Castração/tratamento farmacológico , Neoplasias de Próstata Resistentes à Castração/genética , Receptores Androgênicos/genética , Tioidantoínas/uso terapêutico , Idoso , Idoso de 80 Anos ou mais , DNA Tumoral Circulante/genética , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Intetumumab is a fully human mAb with antiangiogenic, antitumor properties which has shown potential therapeutic effect in castration-resistant prostate cancer (CRPC) patients. PATIENTS AND METHODS: In a phase 2, randomized, double-blind, multicenter study, men with metastatic CRPC without prior systemic nonhormonal therapy were randomly assigned to 75-mg/m(2) docetaxel (Taxotere) and 5-mg prednisone plus placebo (N = 65) or 10-mg/kg intetumumab (N = 66) q3w. Placebo patients with progressive disease (PD) could cross over to 10-mg/kg intetumumab alone or with docetaxel. The primary end-point was progression-free survival (PFS). The secondary end-points included tumor response (complete response + partial response, CR + PR), prostate-specific antigen (PSA) response, and overall survival (OS). RESULTS: All efficacy end-points favored placebo over intetumumab, including PFS (median 11.0 versus 7.6 months, P = 0.014), tumor response (20% versus 16%, P = 0.795), PSA response (68% versus 47%, P = 0.018), OS (median 20.6 versus 17.2 months, P = 0.163). Common all-grade adverse events (AEs) with placebo and intetumumab were alopecia (43% versus 26%); diarrhea, leukopenia (both 34% versus 27%); neutropenia (35% versus 23%). Grade ≥ 3 leukopenia (28% versus 17%) and neutropenia (26% versus 18%) occurred more often with placebo than with intetumumab. Intetumumab serum concentrations increased with repeated dosing and did not reach steady-state. Greater decreases in N-telopeptide of type I collagen (NTx), C-telopeptide (CTx) and CTCs occurred with intetumumab than with placebo. CONCLUSION: The addition of intetumumab to docetaxel resulted in shorter PFS without additional toxicity among CRPC patients.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Integrina alfaV/imunologia , Prednisona/uso terapêutico , Neoplasias da Próstata/tratamento farmacológico , Taxoides/uso terapêutico , Idoso , Inibidores da Angiogênese/uso terapêutico , Anticorpos Monoclonais/efeitos adversos , Anticorpos Monoclonais Humanizados , Antineoplásicos/efeitos adversos , Antineoplásicos/uso terapêutico , Antineoplásicos Hormonais/efeitos adversos , Antineoplásicos Hormonais/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Intervalo Livre de Doença , Docetaxel , Método Duplo-Cego , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica/tratamento farmacológico , Orquiectomia , Placebos/administração & dosagem , Prednisona/efeitos adversos , Neoplasias da Próstata/mortalidade , Sobrevida , Taxoides/efeitos adversosRESUMO
BACKGROUND: α(v) integrins are involved in angiogenesis and melanoma tumourigenesis. Intetumumab (CNTO 95) is a fully human anti-α(v)-integrin monoclonal antibody. METHODS: In a multicentre, randomised, phase II study, stage IV melanoma patients were randomised 1:1:1:1 to 1000 mg m(-2) dacarbazine+placebo (n=32), 1000 mg m(-2) dacarbazine+10 mg kg(-1) intetumumab (n=32), 10 mg kg(-1) intetumumab (n=33), or 5 mg kg(-1) intetumumab (n=32) q3w. The primary endpoint was progression-free survival (PFS). Secondary endpoints included overall survival (OS), objective response rate (ORR), adverse events, and pharmacokinetics. RESULTS: No statistically significant differences in efficacy were observed between groups. In the dacarbazine+placebo, dacarbazine+intetumumab, 10 mg kg(-1) intetumumab, and 5 mg kg(-1) intetumumab groups, median PFS was 1.8, 2.5, 1.4, and 1.4 months; median OS was 8, 11, 15, and 9.8 months; and ORR of complete+partial response was 10, 3, 6, and 0%. Nonlinear intetumumab pharmacokinetics and potential intetumumab-dacarbazine interactions were observed. Transient, asymptomatic, nonrecurring, grade 1-2, uveitic reactions that resolved spontaneously or with topical steroids were seen in 22-30% of intetumumab-treated patients. Low-grade infusion-reaction symptoms (headache, fatigue, nausea, vomiting, fever, chills) were observed, as expected, in 16-73% of dacarbazine-treated patients. No intetumumab-related myelosuppression, laboratory/electrocardiogram abnormalities, or deaths occurred. CONCLUSION: With its favourable safety profile and a nonsignificant trend towards improved OS, intetumumab merits further investigation in advanced melanoma.
Assuntos
Anticorpos Monoclonais/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Dacarbazina/administração & dosagem , Integrina alfaV/imunologia , Melanoma/tratamento farmacológico , Neoplasias Cutâneas/tratamento farmacológico , Idoso , Anticorpos Monoclonais/efeitos adversos , Anticorpos Monoclonais Humanizados , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Melanoma/mortalidade , Pessoa de Meia-Idade , Uveíte/induzido quimicamenteRESUMO
Centrocytic lymphoma (CC) and intermediately differentiated lymphocytic lymphoma (IDL) are B-cell non-Hodgkin's lymphomas composed of lymphocytes presumably derived from follicle mantle cells. In these lymphomas, a specific chromosomal translocation, t(11;14)(q13;q32), has been described. Previous studies suggested an association between t(11;14) chromosomal translocations and BCL-1 rearrangements. To evaluate the association between BCL-1 rearrangements and CC/IDL, Southern blot analysis was performed on a panel of 20 cases of CC/IDL, 22 cases of morphologically similar non-Hodgkin's lymphomas, 11 cases of chronic B-cell leukemias, and 2 cases of myelomas. We used various probes covering a considerable proportion of the 120-kilobase BCL-1 locus, and rearrangements in 50% of CC/IDL (10 of 20) were detected. In CC, all 4 breakpoints were located at the major translocation cluster (MTC). In contrast, in IDL, rearrangements were detected in 3 different cluster regions: 2 cases in the MTC, 2 cases with a breakpoint 24 kilobases outside the MTC, and 2 additional cases with breakpoints found 3 kilobases 5' of the first exon of the PRAD1/CCND1 gene, which is located 120 kilobases outside the MTC. In addition, one leukemia showed a breakpoint 63 kilobases outside the MTC. In all cases, there was comigration of the rearranged 11q13 fragment and the immunoglobulin heavy chain-joining gene complex, indicating a t(11;14)(q13;q32) chromosomal rearrangement. Our results show that Southern blot analysis is helpful to identify CC/IDL, but multiple breakpoints are present over a large region, and therefore, many probes are necessary to cover all breakpoints.
Assuntos
Ciclinas/genética , Rearranjo Gênico , Linfoma de Células B/genética , Proteínas Oncogênicas/genética , Oncogenes , Proteínas Proto-Oncogênicas/genética , Mapeamento Cromossômico , Cromossomos Humanos Par 11 , Ciclina D1 , Genes de Imunoglobulinas , Humanos , Cadeias Pesadas de Imunoglobulinas/genética , Região de Junção de Imunoglobulinas/genéticaRESUMO
In mantle cell lymphoma (MCL) a recurrent chromosomal rearrangement, t(11;14)(q13;q32), has been described. Most breakpoints have been detected within the 120 kb BCL-1 region, upstream of the cyclin D1 gene. To evaluate the association between BCL-1 rearrangement and expression of cyclin D1 in lymphoproliferative disorders, we analysed a series of 24 MCL, 56 other B-cell non-Hodgkin's lymphomas (NHL), 28 chronic B-cell leukemias, 18 hematopoietic cell lines and 10 normal lymphoid tissues at the RNA level. Hematopoietic cell lines with a known 11q13 translocation showed high expression of the 4.5 kb cyclin D1 transcript. Three B-cell lines without known 11q13 breakpoint showed low expression. We detected high expression in all (11/11) MCL with and in 11 out of 13 cases of MCL without detectable t(11;14) rearrangement. In three cases with a rearrangement at the 3' end of cyclin D1, two showed overexpression of the 1.5 kb transcript and one expression of an aberrant (3.0 kb) transcript. In other lymphoproliferative disorders, only 5/15 hairy cell leukemias, all without detectable t(11;14), and 5/8 B-cell leukemias suspected to be MCL in leukemic phase showed expression levels comparable to MCL, whereas no or only low expression were observed in 56 cases of other NHL, seven chronic B-cell leukemias and all (10/10) normal lymphoid tissues. Cell sorting experiments on fresh tonsils showed that this low expression was present in normal B-cells and not in T-cells. In contrast to other studies, our data indicate that cyclin D1 is expressed in many lymphoproliferative disorders and normal tissues, albeit at low levels. High levels of expression of cyclin D1 however is restricted to MCL and some hairy cell leukemias. We therefore propose that overexpression of cyclin D1 is a reliable marker for the classification of MCL.
Assuntos
Ciclinas/genética , Linfoma não Hodgkin/genética , Proteínas Oncogênicas/genética , RNA Neoplásico/genética , Biomarcadores Tumorais , Cromossomos Humanos Par 11 , Cromossomos Humanos Par 14 , Ciclina D1 , Expressão Gênica , Rearranjo Gênico , Humanos , Linfoma não Hodgkin/patologia , Proteínas Proto-Oncogênicas/genética , Proto-Oncogenes , RNA Mensageiro/genética , Translocação GenéticaRESUMO
In mantle cell lymphoma (MCL), in addition to the characteristic t(11;14)(q13;q32), rearrangements on the 3' side of the cyclin D1 gene have been described. In a series of 32 MCL we found three cases with 3'-rearrangements by Southern blot analysis with a probe representing the 3' untranslated region of the cyclin D1 gene. All three were characterized by the absence of the 4.5 kb transcript, and instead, two cases showed overexpression of the 1.7 kb and a third case (MCLp14) an aberrant 2.5 kb transcript. At the genomic level, fine mapping experiments showed in the 3' untranslated region a 2 kb deletion in MCLp14 and a breakpoint in the other two cases. Fiber FISH analysis showed that in all three cases the 3' aberration co-exists with a regular t(11;14) breakpoint 5' of cyclin D1 on a single allele. Fiber FISH analysis of 16 additional MCL revealed two other such cases with breakpoints 5' and 3' of cyclin D1. These mono-allelic aberrations affecting the cyclin D1 gene suggest that the t(11;14) as a first step switches on transcription, and then sequences in the untranslated region of cyclin D1 are affected to stabilize the message.
Assuntos
Aberrações Cromossômicas , Ciclina D1/genética , Linfoma não Hodgkin/genética , Sequência de Bases , Southern Blotting , Quebra Cromossômica , Cromossomos Humanos Par 11 , Cromossomos Humanos Par 14 , Humanos , Hibridização in Situ Fluorescente/métodos , Mapeamento por Restrição , Translocação GenéticaRESUMO
PURPOSE: From a population-based non-Hodgkin's lymphoma (NHL) registry, 41 patients with mantle cell lymphoma (MCL) -- a recently defined distinct B-cell NHL -- were selected and compared with patients with low- or intermediate-grade NHL from the same registry. PATIENTS AND METHODS: The incidence and behavior of MCL in the area of the Comprehensive Cancer Center West (CCCW) from 1981 to 1989 were analyzed. Age, performance, tumor bulk, extranodal localization, stage, response to therapy, and survival were registered. Expression of cyclin D1 protein and Ki-67 were measured in 29 patients. RESULTS: MCL made up 3.7% of NHLs. The median age was 68 years, and the male-to-female ratio was 1.6:1. Seventy-eight percent presented with stage IV, with the majority having bone marrow involvement. The complete response (CR) rate was 32% (13 of 41), with a median duration of 25 months. The median overall survival time was 31.5 months. The International Prognostic Index identified five patients with a low-risk score and a median survival time of 93+ months. In 23 of 29 patients, cyclin D1 overexpression was present, without any relation to overall or disease-free survival. In contrast, a proliferative index less than 10% was significantly related to a better overall survival time (50 v 24 months). CONCLUSION: MCL is a disease of the elderly, who present with widespread disease and with a poor response to therapy. Although it harbors features of an indolent NHL, it behaves clinically as an aggressive NHL with a short overall survival time.
Assuntos
Linfoma não Hodgkin/epidemiologia , Linfoma não Hodgkin/patologia , Idoso , Divisão Celular , Feminino , Humanos , Imuno-Histoquímica , Incidência , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Países Baixos/epidemiologia , Prognóstico , Sistema de Registros , Fatores de Risco , Análise de SobrevidaRESUMO
The glycoprotein recognized by the monoclonal antibody (mAb) 17-1A is present on most carcinomas, which makes it an attractive target for immunotherapy. Indeed, adjuvant treatment with mAb 17-1A did successfully reduce the 5 years mortality among colorectal cancer patients with minimal residual disease. Currently the antibody is approved for clinical use in Germany, and is on its way to approval in a number of other countries. New immunotherapeutic strategies targeting the 17-1A antigen are in development or even in early-phase clinical trials. Therefore, a better understanding of the biology of the 17-1A antigen may result in improved strategies for the treatment and diagnosis of human carcinomas. In this review the properties of the 17-1A antigen are discussed concerning tumor biology and the function of the molecule. This 40-kDa glycoprotein functions as an Epithelial Cell Adhesion Molecule, therefore the name Ep-CAM was suggested. Ep-CAM mediates Ca2+-independent homotypic cell-cell adhesions. Formation of Ep-CAM-mediated adhesions has a negative regulatory effect on adhesions mediated by classic cadherins, which may have strong effects on the differentiation and growth of epithelial cells. Indeed, in vivo expression of Ep-CAM is related to increased epithelial proliferation and negatively correlates with cell differentiation. A regulatory function of Ep-CAM in the morphogenesis of epithelial tissue has been demonstrated for a number of tissues, in particular pancreas and mammary gland. The function of Ep-CAM should be taken into consideration when developing new therapeutic approaches targeting this molecule.
Assuntos
Antígenos de Neoplasias/fisiologia , Carcinoma/imunologia , Moléculas de Adesão Celular/fisiologia , Adulto , Animais , Anticorpos Monoclonais/uso terapêutico , Antígenos de Neoplasias/análise , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/imunologia , Biomarcadores Tumorais/análise , Mama/química , Mama/embriologia , Caderinas/fisiologia , Carcinoma/química , Carcinoma/diagnóstico , Carcinoma/terapia , Adesão Celular/fisiologia , Moléculas de Adesão Celular/análise , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/imunologia , Diferenciação Celular , Transformação Celular Neoplásica , Cromossomos Humanos Par 4/genética , Molécula de Adesão da Célula Epitelial , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Evolução Molecular , Feminino , Proteínas Fetais/fisiologia , Regulação Neoplásica da Expressão Gênica , Genes , Humanos , Imuno-Histoquímica , Imunoterapia , Masculino , Camundongos , Camundongos Transgênicos , Morfogênese , Família Multigênica , Especificidade de Órgãos , Pâncreas/química , Pâncreas/embriologia , Conformação Proteica , Estrutura Terciária de ProteínaRESUMO
In order to identify helpful markers in the classification of mantle cell lymphoma, a morphological, immunohistochemical and molecular genetic analysis of 41 cases of NHL, originally referred to us as CC, ILL or IDL, was performed. We revised these lymphomas using the strict morphological criteria described in the updated Kiel classification and the more recently described criteria for MCL. The term MCL was used to designate the small lymphocytic B-cell NHL, previously referred to as CC or ILL/ IDL. This revision yielded 20 MCL, 8 CLL, 3 Cb/Cc, 1 CB, 6 IC and 3 MALT lymphomas. The presence of scattered histiocytes was seen in 90% of MCL and 5% of the other cases. No other morphological parameter, besides the used criteria, differentiated between MCL and similar small lymphocytic B cell lymphomas. Helpful immunohistochemical markers to distinguish MCL from similar small lymphocytic lymphomas were CD5+, CD10-, CD23- and Alkaline Phosphatase+. Large fields of dendritic reticulum cells, often in a loose and disrupted arrangement were seen in 82% of MCL and in 19% of the other lymphomas. Analysis with Southern blotting showed a rearrangement in the BCL-1 locus in 12/20 cases of MCL but not in the other 21 lymphomas. Although very specific for MCL, Southern blotting to detect BCL-1 rearrangements is, due to the large number of probes necessary, not of great help in daily practice for routine diagnostic purposes. We conclude that using strict morphological criteria, the diagnosis MCL can be made reliably and that immunophenotyping is helpful in supporting the diagnosis.
Assuntos
Imuno-Histoquímica , Linfoma de Células B/diagnóstico , Linfoma de Células B/genética , Linfoma não Hodgkin/imunologia , Linfoma não Hodgkin/patologia , Biologia Molecular , Humanos , Linfoma de Células B/classificação , Linfoma de Células B/imunologia , Linfoma de Células B/patologia , Linfoma não Hodgkin/diagnóstico , Linfoma não Hodgkin/genéticaAssuntos
Febre Suína Africana/imunologia , Doença Aguda , África , Febre Suína Africana/complicações , Animais , Portador Sadio/microbiologia , Portador Sadio/veterinária , Doença Crônica , Testes de Fixação de Complemento , Surtos de Doenças/veterinária , Hipergamaglobulinemia/complicações , Hipergamaglobulinemia/veterinária , Testes de Precipitina , Suínos , Vacinas Virais/administração & dosagemRESUMO
Mantle-cell lymphoma comprises 2%-10% of all non-Hodgkin's lymphomas (NHLs). Patients present with generalized disease, and have a poor prognosis. Three different histologic patterns (mantle zone, nodular, and diffuse) and three different cytological variants (classical, blastic, and pleomorphic) have been described. The phenotype (strong surface IgM, CD5+, CD10-, CD23-, cyclin D1+ and B-cell markers+) is remarkably constant. Dependent on the methods used (PCR, Southern blot analysis, and cytogenetics) a t(11;14) can be detected in approximately 35%-66% of cases. Using FISH analysis, possibly almost all cyclin D1-expressing MCLs carry this translocation, indicating that a substantial part of these translocations are missed by conventional methods. This has been confirmed by DNA fiber FISH analysis by which the breakpoints could be accurately mapped over a 220 kb region centromeric of the cyclin D1 gene. Additional genetic abnormalities involve breakpoints and deletion at the 3' end of the cyclin D1 gene, numerical chromosomal aberrations, mutations in p53, and deletions of p16. These may be associated with tumor progression. Owing to the translocation t(11;14), the cyclin D1 gene is activated. At the RNA level, approximately 90% of MCLs show overexpression. This corroborates immunohistochemistry on paraffin tissue sections. Since expression of cyclin D1 in normal lymphoid cells is very low to undetectable, and only hairy-cell leukemia and very few other B-cell lymphomas show expression, immunohistochemistry for cyclin D1 provides an excellent marker for MCL. In hairy-cell leukemia, expression is moderate and cannot be explained by chromosomal translocation.
Assuntos
Biomarcadores Tumorais/análise , Ciclinas/análise , Linfoma não Hodgkin/química , Proteínas Oncogênicas/análise , Cromossomos Humanos Par 11 , Cromossomos Humanos Par 14 , Ciclina D1 , Demografia , Humanos , Imunofenotipagem , Linfoma não Hodgkin/genética , Linfoma não Hodgkin/patologia , Translocação GenéticaRESUMO
Thirty-three pigs in three groups of nineteen, ten, and four pigs were infected with three different African swine fever (ASF) virus isolates, respectively. All virus isolates were attenuated to varying degrees by passaging in cell cultures, and they retained sufficiently low virulence to produce subacute and chronic infections in pigs. Sera collected at various intervals were tested for antibody activity by the immunoelectroosmophoresis, agar gel diffusion precipitin, and complement-fixation tests using a modified Kolmer technique. Results clearly indicated that the immunoelectroosmophoresis test is a rapid (30 minute) and accurate method with extreme sensitivity and superior to the complement-fixation and agar gel diffusion precipitin tests in detecting antibody against ASF virus. Possible use of this method in detecting ASF virus infection is suggested.
Assuntos
Febre Suína Africana/imunologia , Anticorpos/análise , Eletroforese das Proteínas Sanguíneas , Febre Suína Africana/diagnóstico , Animais , Antígenos Virais , Células Cultivadas , Testes de Fixação de Complemento , Epitopos , Haplorrinos/imunologia , Soros Imunes , Imunodifusão , Rim , Métodos , SuínosRESUMO
Ep-CAM is a homophilic, Ca2+-independent cell-cell adhesion molecule that is expressed in many human epithelial tissues. Its increased expression is closely associated with active cell proliferation. Furthermore, in epithelial cell types that in adults lack Ep-CAM (i. e. squamous epithelia), up-regulation of Ep-CAM coincides with the early stages of neoplastic change. This study has analysed the expression of Ep-CAM in liver, in the hepatocytes and cells of the biliary duct system, in relation to proliferative diseases and carcinogenesis. Adult hepatocytes are Ep-CAM negative, with only bile duct epithelium being positive in the liver tissue. However, in the 8-week embryonic liver, the majority of hepatocytes express Ep-CAM. During regeneration and repair of liver tissues associated with focal nodular hyperplasia and (biliary) cirrhosis, activation of Ep-CAM expression was observed, with high expression levels in so-called 'ductular proliferations'-regenerating stem cells. During precursor cell differentiation into mature hepatocytes, several intermediate morphological stages could be observed, all Ep-CAM positive, including cells morphologically close to mature hepatocytes. Full maturation of the precursor resulted in the disappearance of Ep-CAM expression. The results suggest that expression of Ep-CAM is a prerequisite of the proliferative phenotype during differentiation of hepatocyte precursors. In liver neoplasia, Ep-CAM was expressed in almost all cholangiocarcinomas (10/11), whereas the majority of hepatocellular carcinomas (8/10) were negative, suggesting that malignant proliferation of hepatocellular carcinoma cells is not related to expression of Ep-CAM and that hepatocellular carcinoma originates from a highly differentiated precursor. The results indicate that Ep-CAM can be used as an additional immunohistochemical marker to distinguish cholangiocarcinoma from hepatocellular carcinoma due to the differential expression of Ep-CAM in these tumours.
Assuntos
Antígenos de Neoplasias/análise , Biomarcadores Tumorais/análise , Carcinoma Hepatocelular/química , Moléculas de Adesão Celular/análise , Neoplasias Hepáticas/química , Fígado/química , Adulto , Neoplasias dos Ductos Biliares/química , Ductos Biliares Intra-Hepáticos , Colangiocarcinoma/química , Molécula de Adesão da Célula Epitelial , Humanos , Imuno-Histoquímica , Fígado/embriologia , Fígado/patologia , Regeneração Hepática , MetaplasiaRESUMO
Mantle cell lymphoma (MCL) is a clinicopathologic entity that is difficult to diagnose on histopathologic criteria. Approximately 50% to 70% of MCL contain a t(11;14)(q13;q32) translocation involving the cyclin D1 gene. Irrespective of this rearrangement, almost all MCL show overexpression of the cyclin D1 gene at the mRNA level. Other B-cell non-Hodgkin's lymphomas (NHL) do not show this rearrangement or overexpression of cyclin D1. We developed an immunohistochemical assay to detect overexpression of the cyclin D1 protein on conventional formalin-fixed, paraffin-embedded biopsies using the well-defined monoclonal antibody DCS-6. Expression in tumor cells was compared with expression of cyclin D1 in endothelial cells and fibroblasts. An exclusively nuclear staining pattern was observed. Moreover, expression was directly compared with the expression observed by immunoblot analysis with the same antibody, as well as with mRNA expression and with the occurrence of genomic rearrangements within the BCL-1 locus. Of 13 MCL that were analyzed by immunohistochemistry and immunoblot, 12 showed overexpression with both techniques, whereas no overexpression was observed in 39 other NHL. Of 13 additional MCL studied either by immunohistochemistry or immunoblot, 11 also showed overexpression. Two lymphomas morphologically indistinguishable from MCL but with an aberrant immunophenotype (CD5 negative, CD10 positive) both lacked overexpression of cyclin D1. These results underscore the significance of overexpression of the cyclin D1 protein as a specific marker for MCL. Detection of cyclin D1 overexpression on formalin-fixed, paraffin-embedded tissues using the DCS-6 monoclonal antibody can be applied for routine diagnostic purposes.
Assuntos
Biomarcadores Tumorais/análise , Ciclinas/análise , Linfoma não Hodgkin/diagnóstico , Proteínas Oncogênicas/análise , Anticorpos Monoclonais , Biópsia , Northern Blotting , Núcleo Celular/química , Ciclina D1 , Ciclinas/genética , Expressão Gênica , Humanos , Immunoblotting , Imuno-Histoquímica , Linfoma não Hodgkin/metabolismo , Proteínas Oncogênicas/genética , Proteínas Proto-Oncogênicas/genética , RNA Mensageiro/metabolismo , Inclusão do Tecido , Translocação GenéticaRESUMO
BACKGROUND: Previous results suggested increased mRNA expression of CCND1 in hairy cell leukemia (HCL). The CCND1 gene is involved in the t(11;14)(q13;q32) chromosomal rearrangement, a characteristic abnormality in mantle cell lymphoma (MCL). We and others reported that, in contrast to other B-cell lymphomas, almost all MCL have over-expression of the CCND1 gene with a good correlation between RNA and protein analysis. Recent studies showed that overexpression of the cyclin D1 protein can be easily detected by immunohistochemistry (IHC) on formalin-fixed, paraffin embedded tissues. PATIENTS AND METHODS: To investigate whether the CCND1 gene is involved in HCL, we performed IHC on a series of 22 cases using formalin-fixed paraffin embedded splenectomy specimens. For IHC the sections were boiled in citrate buffer. The presence of rearrangements within the BCL-1 locus and the CCND1 gene was analyzed in 13 of 22 cases by Southern blot analysis using all available break-point probes. Expression of CCND1 was analyzed at the mRNA level (Northern blot) and protein level (IHC). RESULTS: Overexpression of the cyclin D1 protein using IHC was observed in all cases, with strong expression in 5 cases. Pre-existing B- and T-cell areas of the spleen did not express significant levels of the cyclin D1 protein. Seven of 9 cases analyzed by both IHC and Northern blotting showed overexpression of the CCND1 gene with both methods. No genomic abnormalities were observed in any of the 13 cases studied by Southern blot analysis. Additionally, no 11q13 abnormalities were detected by banding analysis of 19 of 22 cases. CONCLUSIONS: The elevated levels of CCND1 mRNA and protein in conjunction with the absence of overt rearrangements within the BCL-1 locus distinguish HCL from MCL and other B-cell malignancies. This suggests that activation of the CCND1 gene in HCL is due to mechanisms other than chromosomal rearrangement.
Assuntos
Ciclinas/análise , Regulação Leucêmica da Expressão Gênica/fisiologia , Rearranjo Gênico , Leucemia de Células Pilosas/genética , Proteínas Oncogênicas/análise , Translocação Genética , Adulto , Idoso , Ciclina D1 , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-IdadeRESUMO
Uterine cervix represents a convenient model for the study of the gradual transformation of normal squamous epithelium via low- to high-grade squamous intraepithelial lesions (SILs). Because SIL, on the basis of the cytokeratins expressed, are thought to originate from the reserve cells, we analyzed whether SILs also show a reserve cell phenotype with respect to intercellular interactions. The changes in expression and subcellular localization of the components of the adherens junction and desmosomal complexes were investigated in normal, metaplastic, and premalignant cervical epithelium, as well as in cell cultures derived from these tissues. The results suggest that 1) during progression of SILs, E-cadherin is suppressed, with its role in cell-cell connections diminishing; 2) P-cadherin, in contrast, becomes the predominant cadherin in high-grade SILs; 3) the level of cellular alpha-catenin is dramatically decreased in high-grade SILs; 4) the level of beta-catenin is decreased during progression of SILs, with plakoglobin suggestively becoming the predominant catenin mediating connection of cadherins to the cytoskeleton; 5) the assembly of desmosomes is affected during progression of SILs and is accompanied by a dramatically decreased expression for desmogleins and desmoplakins (I, II); and 6) expression of differentiation markers (involucrin, CK13) in high-grade SILs seems to be controlled by P-cadherin as opposed to E-cadherin in the normal tissue counterpart. We conclude that during development of cervical lesions substantial (both quantitative and qualitative) changes occur in cell-cell junctions, making the interactions of cells in lesions dissimilar from those of reserve cells, basal cells, or cells of immature squamous metaplasia, despite existing morphological similarity between all of these cell types and cells of high-grade lesions.
Assuntos
Caderinas/fisiologia , Proteínas do Citoesqueleto/fisiologia , Transativadores , Displasia do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/metabolismo , Biomarcadores , Caderinas/análise , Moléculas de Adesão Celular/análise , Moléculas de Adesão Celular/fisiologia , Células Cultivadas , Proteínas do Citoesqueleto/análise , Desmogleínas , Desmoplaquinas , Desmossomos/metabolismo , Progressão da Doença , Feminino , Quinase 1 de Adesão Focal , Proteína-Tirosina Quinases de Adesão Focal , Humanos , Imuno-Histoquímica , Proteínas Tirosina Quinases/análise , Proteínas Tirosina Quinases/fisiologia , Fatores de Tempo , Neoplasias do Colo do Útero/diagnóstico , alfa Catenina , beta Catenina , gama CateninaRESUMO
Several hematologic malignancies are associated with specific chromosomal translocations. Because of the dispersed distribution, chromosomal breakpoints may be difficult to detect using molecular techniques. We present a new application of a recently developed method, DNA fiber fluorescence in situ hybridization (fiber FISH), which allows direct visualization and mapping of chromosomal breakpoints. We tested this method for detection of the t(11;14)(q13;q32) translocation in mantle cell lymphoma. In DNA fiber FISH, a series of fluorochrome-labeled DNA probes covering several hundreds of kilobasepairs is hybridized to linear DNA molecules (or fibers) prepared from frozen tissue or intact cells. By using alternate fluorescent colors, a potential breakpoint region is stained in a color barcode pattern. Breaks in this region will split the barcode in two complementary parts, from which the breakpoint position can be derived. We used a 250-kb barcode covering the BCL-1 locus to detect 11q13 breakpoints in 20 well-characterized mantle cell lymphomas. A t(11;14) was shown by cohybridization of these probes with probes for the Ig heavy chain locus at 14q32. In 18 of 20 mantle cell lymphomas, a breakpoint within the 11q13/BCL-1 barcode was shown by the presence of multiple, complementary translocation products. Fusion of 11q13 and 14q32 sequences on single fibers indicating t(11;14)(q13;q32) was found in all 18 breakpoint-positive mantle cell lymphomas. In one additional case, fusion of an intact 11q13 barcode with 14q32 sequences indicated a breakpoint 100 kb centromeric of the major translocation cluster of BCL-1. Within the 120-kb region of BCL-1, breakpoints were widely scattered. This explains why, so far, a BCL-1 breakpoint had been detected by Southern blot analysis in only 10 of 19 cases. DNA fiber FISH analysis showed a t(11;14) in 95% of mantle cell lymphoma. The results indicate that DNA fiber FISH is a rapid, simple, and equally powerful method for detection of clustered and dispersed translocation breakpoints.