RESUMO
OBJECTIVES: To describe the CT appearance of anal sac adenocarcinoma lesions in a population of dogs including the relations between primary tumour, and locoregional and distant metastasis. MATERIALS AND METHODS: Retrospective review of dogs with confirmed anal sac adenocarcinoma and available CT images of the thorax, abdomen and pelvis. RESULTS: A population of 70 dogs were included in the study. No association was found between anal sac mass size and presence or absence of iliosacral lymph node enlargement. The prevalence of local metastatic disease characterised by iliosacral lymphadenomegaly in this study was 71%, with pulmonary metastases identified in 11% of cases. There were no cases of distant pulmonary metastasis without concurrent locoregional lymphadenomegaly. CLINICAL SIGNIFICANCE: In our population of dogs local metastatic spread of anal sac adenocarcinoma was common, with a relatively low prevalence of pulmonary metastasis. The study demonstrates the importance of thorough rectal examination and/or imaging to assess the iliosacral lymph centre in this disease irrespective of the size of the anal sac mass.
Assuntos
Adenocarcinoma , Neoplasias das Glândulas Anais , Sacos Anais , Doenças do Cão , Adenocarcinoma/diagnóstico por imagem , Adenocarcinoma/patologia , Adenocarcinoma/veterinária , Neoplasias das Glândulas Anais/diagnóstico por imagem , Sacos Anais/diagnóstico por imagem , Sacos Anais/patologia , Animais , Doenças do Cão/diagnóstico , Cães , Estadiamento de Neoplasias , Estudos Retrospectivos , Tomografia Computadorizada por Raios X/veterináriaRESUMO
The hyperparathyroidism-jaw tumour (HPT-JT) syndrome is an autosomal dominant disorder characterized by occurrence of parathyroid tumours, often atypical adenomas and carcinomas, ossifying jaw fibromas, renal tumours and uterine benign and malignant neoplasms. HPT-JT is caused by mutations of the cell division cycle 73 (CDC73) gene, located on chromosome 1q31.2 and encodes a 531 amino acid protein, parafibromin. To facilitate in vivo studies of Cdc73 in tumourigenesis we generated conventional (Cdc73+/-) and conditional parathyroid-specific (Cdc73+/L/PTH-Cre and Cdc73L/L/PTH-Cre) mouse models. Mice were aged to 18-21 months and studied for survival, tumour development and proliferation, and serum biochemistry, and compared to age-matched wild-type (Cdc73+/+ and Cdc73+/+/PTH-Cre) littermates. Survival of Cdc73+/- mice, when compared to Cdc73+/+ mice was reduced (Cdc73+/-=80%; Cdc73+/+=90% at 18 months of age, P<0.05). Cdc73+/-, Cdc73+/L/PTH-Cre and Cdc73L/L/PTH-Cre mice developed parathyroid tumours, which had nuclear pleomorphism, fibrous septation and increased galectin-3 expression, consistent with atypical parathyroid adenomas, from 9 months of age. Parathyroid tumours in Cdc73+/-, Cdc73+/L/PTH-Cre and Cdc73L/L/PTH-Cre mice had significantly increased proliferation, with rates >fourfold higher than that in parathyroid glands of wild-type littermates (P<0.0001). Cdc73+/-, Cdc73+/L/PTH-Cre and Cdc73L/L/PTH-Cre mice had higher mean serum calcium concentrations than wild-type littermates, and Cdc73+/- mice also had increased mean serum parathyroid hormone (PTH) concentrations. Parathyroid tumour development, and elevations in serum calcium and PTH, were similar in males and females. Cdc73+/- mice did not develop bone or renal tumours but female Cdc73+/- mice, at 18 months of age, had uterine neoplasms comprising squamous metaplasia, adenofibroma and adenomyoma. Uterine neoplasms, myometria and jaw bones of Cdc73+/- mice had increased proliferation rates that were 2-fold higher than in Cdc73+/+ mice (P<0.05). Thus, our studies, which have established mouse models for parathyroid tumours and uterine neoplasms that develop in the HPT-JT syndrome, provide in vivo models for future studies of these tumours.
Assuntos
Adenoma/genética , Carcinoma/genética , Fibroma/genética , Hiperparatireoidismo/genética , Neoplasias Maxilomandibulares/genética , Neoplasias das Paratireoides/genética , Proteínas Supressoras de Tumor/genética , Neoplasias Uterinas/genética , Adenoma/complicações , Animais , Carcinoma/complicações , Feminino , Fibroma/complicações , Deleção de Genes , Hiperparatireoidismo/complicações , Neoplasias Maxilomandibulares/complicações , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neoplasias das Paratireoides/complicações , Neoplasias Uterinas/complicaçõesRESUMO
Parafibromin is a nuclear protein with a tumour suppressor role in the development of non-hereditary and hereditary parathyroid carcinomas, and the hyperparathyroidism-jaw tumour (HPT-JT) syndrome, which is associated with renal and uterine tumours. Nuclear localization signal(s), (NLS(s)), of the 61 kDa parafibromin remain to be defined. Utilization of computer-prediction programmes, identified five NLSs (three bipartite (BP) and two monopartite (MP)). To investigate their functionality, wild-type (WT) and mutant parafibromin constructs tagged with enhanced green fluorescent protein or cMyc were transiently expressed in COS-7 cells, or human embryonic kidney 293 (HEK293) cells, and their subcellular locations determined by confocal fluorescence microscopy. Western blot analyses of nuclear and cytoplasmic fractions from the transfected cells were also performed. WT parafibromin localized to the nucleus and deletions or mutations of the three predicted BP and one of the predicted MP NLSs did not affect this localization. In contrast, deletions or mutations of a MP NLS, at residues 136-139, resulted in loss of nuclear localization. Furthermore, the critical basic residues, KKXR, of this MP NLS were found to be evolutionarily conserved, and over 60% of all parafibromin mutations lead to a loss of this NLS. Thus, an important functional domain of parafibromin, consisting of an evolutionarily conserved MP NLS, has been identified.
Assuntos
Sinais de Localização Nuclear , Proteínas Nucleares/química , Proteínas Supressoras de Tumor/química , Sequência de Aminoácidos , Animais , Western Blotting , Células COS , Núcleo Celular/química , Chlorocebus aethiops , Sequência Conservada , Evolução Molecular , Proteínas de Fluorescência Verde/análise , Proteínas de Fluorescência Verde/genética , Humanos , Dados de Sequência Molecular , Mutação , Proteínas Nucleares/análise , Proteínas Nucleares/genética , Estrutura Terciária de Proteína , Alinhamento de Sequência , Proteínas Supressoras de Tumor/análise , Proteínas Supressoras de Tumor/genéticaRESUMO
OBJECTIVE: To investigate two patients with the hyperparathyroidism-jaw tumour (HPT-JT) syndrome and three patients with familial isolated hyperparathyroidism (FIHP), together with 31 parathyroid tumours (2 HPT-JT, 2 FIHP and 27 sporadic) for HRPT2 mutations. The HPT-JT syndrome and FIHP are autosomal dominant disorders that may be caused by abnormalities of the HRPT2 gene, located on chromosome 1q31.2. HRPT2 encodes a 531 amino acid protein, parafibromin, which interacts with human homologues of the yeast Paf1 complex. DESIGN: Leukocyte and tumor DNA was used with HRPT2-specific primers for polymerase chain reaction amplification of the 17 exons and their splice junctions, and the DNA sequences of the polymerase chain reaction products determined. RESULTS: Three heterozygous germline HRPT2 mutations, two in HPT-JT and one in FIHP patients, were identified. These consisted of one 1-bp duplication (745dup1bp), 1 nonsense (Arg234Stop) and 1 missense (Asp379Asn) mutation. One parathyroid tumour from an FIHP patient was demonstrated to harbour a germline deletion of 1 bp together with a somatic missense (Leu95Pro) mutation, consistent with a 'two-hit' model for hereditary cancer. The 27 sporadic benign parathyroid tumours did not harbour any HRPT2 somatic mutations. Six HRPT2 polymorphisms with allele frequencies ranging from 2% to 15% were detected. CONCLUSIONS: Our results have identified three novel HRPT2 mutations (two germline and one somatic). The Asp379Asn mutation is likely to disrupt interaction with the human homologue of the yeast Paf1 complex, and the demonstration of combined germline and somatic HRPT2 mutations in a parathyroid tumour provide further evidence for the tumour suppressor role of the HRPT2 gene.
Assuntos
Hiperparatireoidismo/genética , Neoplasias Maxilomandibulares/genética , Neoplasias das Paratireoides/genética , Proteínas Supressoras de Tumor/genética , Adulto , Criança , DNA de Neoplasias/genética , Saúde da Família , Feminino , Frequência do Gene , Humanos , Perda de Heterozigosidade/genética , Masculino , Pessoa de Meia-Idade , Mutação/genética , Linhagem , Polimorfismo Genético/genética , SíndromeRESUMO
More than 99% of all splice sites conform to consensus sequences that usually include the invariant dinucleotides gt and ag at the 5' and 3' ends of the introns, respectively. We report on the utilisation of a non-consensus (non-canonical) donor splice site within exon 1 of the HRPT2 gene in familial isolated primary hyperparathyroidism (FIHP). HRPT2 mutations are more frequently associated with the hyperparathyroidism-jaw tumour syndrome (HPT-JT). Patients with FIHP were identified to have a donor splice site mutation, IVS1+1 g-->a, and the consequences of this for RNA processing were investigated. The mutant mRNA lacked 30 bp and DNA sequence analysis revealed this to result from utilisation of an alternative cryptic non-canonical donor splice site (gaatgt) in exon 1 together with the normally occurring acceptor splice site in intron 1. Translation of this mutant mRNA predicted the in-frame loss of 10 amino acids in the encoded protein, termed PARAFIBROMIN. Thus, these FIHP patients are utilising a ga-ag splice site pair, which until recently was considered to be incompatible with splicing but is now known to occur as a rare (<0.02%) normal splicing variant.
Assuntos
Processamento Alternativo , Hiperparatireoidismo Primário/genética , Sítios de Splice de RNA , Proteínas Supressoras de Tumor/genética , Adulto , Análise Mutacional de DNA , Feminino , Humanos , Mutação , Proteínas Proto-Oncogênicas/genética , RNA Mensageiro/metabolismoRESUMO
The hyperparathyroidism-jaw tumour (HPT-JT) syndrome is an autosomal dominant disorder characterized by parathyroid tumours, which are frequently carcinomas, and ossifying jaw fibromas. In addition, some patients may develop renal tumours and cysts. The gene causing HPT-JT, which is referred to as HRPT2 and is located on chromosome 1q31.2, encodes a 531 amino acid protein called PARAFIBROMIN. To date 42 mutations, of which 22 are germline, have been reported and 97% of these are inactivating and consistent with a tumour suppressor role for HRPT2. We have investigated another four HPT-JT families for germline mutations, searched for additional clinical phenotypes, and examined for a genotype-phenotype correlation. Mutations were found in two families. One family had a novel deletional-insertion at codon 669, and the other had a 2 bp insertion at codon 679, which has been reported in four other unrelated patients. These five unrelated patients and their families with the same mutation were not found to develop the same tumours, thereby indicating an absence of a genotype-phenotype correlation. An analysis of 33 HPT-JT kindreds revealed that affected women in 13 HPT-JT families suffered from menorrhagia in their second to fourth decades. This often required hysterectomy, which revealed the presence of uterine tumours. This resulted in a significantly reduced maternal transmission of the disease. Thus, the results of our analysis expand the spectrum of HPT-JT-associated tumours to include uterine tumours, and these may account for the decreased reproductive fitness in females from HPT-JT families.
Assuntos
Hiperparatireoidismo/genética , Neoplasias Maxilomandibulares/genética , Neoplasias Primárias Múltiplas/genética , Neoplasias Uterinas/genética , Adulto , Saúde da Família , Feminino , Genótipo , Humanos , Hiperparatireoidismo/patologia , Neoplasias Maxilomandibulares/patologia , Masculino , Menorragia/complicações , Menorragia/patologia , Pessoa de Meia-Idade , Mutação , Neoplasias Primárias Múltiplas/patologia , Fenótipo , Proteínas/genética , Síndrome , Proteínas Supressoras de Tumor , Neoplasias Uterinas/patologiaRESUMO
OBJECTIVES: Anecdotal reports have suggested that silent corticotroph tumours behave in an aggressive fashion; however, clear comparative data with other non-functioning adenomas (NFAs) are lacking. The aims of the study were, first, to review the natural history of those non-functioning pituitary adenomas with positive immunoreactivity for ACTH and secondly, to determine whether this subgroup behave more aggressively than ACTH immunonegative NFAs by means of comparison with existing departmental data. METHODS AND PATIENTS: Twenty-eight patients (16 men, mean age 51.3 years) who underwent transsphenoidal surgery in Oxford between 1975 and 2001 for clinically non-functioning adenomas where the subsequent immunostaining was positive for ACTH were identified from the patient database. All patients with silent corticotroph tumours who presented during this time period have been included in the analysis; three of the patients have subsequently died but none have been lost to follow-up. The mean follow-up period was 7.4 years (range 0.5-26.9 years) and the results were compared with departmental data for NFAs which were immunonegative for ACTH. None of the patients had clinical evidence of Cushing's syndrome. Tumour invasiveness was classified according to the modified Hardy criteria (Grade 1 = microadenoma (< 1 cm), Grade 2 = macroadenoma (> 1 cm) +/- suprasellar extension, Grade 3 = local invasion with bony destruction and tumour in sphenoid/cavernous sinus and Grade 4 = central nervous system (CNS) spread or extracranial spread, i.e. metastatic). Tumour recurrence was defined as an increase in tumour size compared with the first postoperative scan which was used as a baseline. Visual field defects were documented in 79% of the 28 patients at presentation compared to 69% in the non-functioning population as a whole (P = 0.3). The preoperative imaging in the silent corticotroph group (13 CT, 14 MRI and one air encephalogram) revealed 68% Grade 2 and 32% Grade 3 adenomas. RESULTS: The recurrence rate in the ACTH immunopositive tumours was 32% at a mean of 5.8 years (range 1-16 years) which was not significantly different from the 33% recurrence rate previously recorded in the ACTH immunonegative tumours (P = 0.9). Two of the patients with silent corticotroph adenomas have suffered multiple recurrences; one patient has had three operations and two courses of radiotherapy for two episodes of recurrence and one patient has had four operations, two courses of radiotherapy and gamma knife therapy after three recurrences in total. In contrast, no patient with an ACTH immunonegative tumour has required more than one course of treatment for tumour regrowth. CONCLUSIONS: This is the first single-centre comparative study of a series of clinically silent ACTH immunopositive tumours and has demonstrated that although they do not recur more often than ACTH immunonegative tumours, when they do regrow they show a more aggressive course. The practical implication of this is that there is no evidence for different postoperative imaging and radiotherapy protocols for ACTH immunopositive and immunonegative NFAs at initial presentation. However, if regrowth of a silent corticotroph tumour does occur then very careful monitoring is essential, after further treatment.
Assuntos
Adenoma/química , Hormônio Adrenocorticotrópico/análise , Biomarcadores Tumorais/análise , Neoplasias Hipofisárias/química , Adenoma/patologia , Adenoma/terapia , Hormônio Adrenocorticotrópico/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/imunologia , Terapia Combinada , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Recidiva Local de Neoplasia , Neoplasias Hipofisárias/patologia , Neoplasias Hipofisárias/terapiaRESUMO
The superficial supramammary lymph nodes of 54 lactating dairy cows were examined ultrasonographically with a 7.5 MHz linear transducer; each node was measured in two planes within 24 hours of recording the milk somatic cell count. In most cows, the nodes were well demarcated from the surrounding tissue. The parenchyma of the nodes ranged from hypoechoic to anechoic, with a central bright hyperechoic area, and a thin hyperechoic line surrounded the nodes. The size of the nodes varied, but their internal architecture remained relatively consistent. Their mean length was 7.4 cm (range 3.5 to 15 cm) and their mean depth was 2.5 cm (range 1.2 to 5.7 cm). They were significantly larger in cows with more lactations (P<0.05), but there were no correlations between their size and either the time calved or the milk somatic cell count. The lymph nodes on sides which were positive in a California milk test were significantly larger than those on sides which were negative (P<0.05).