RESUMO
OBJECTIVE: Our aim was to characterize the molecular and genotypic profile of eight thyroid carcinoma-derived cell lines-TPC1, FB2, B-CPAP, K1, XTC-1, C643, 8505C, and Hth74-in order to use them as in vitro models of thyroid carcinogenesis. DESIGN: We evaluated the expression of five thyroid-specific genes (Tg, TSHr, TPO, PAX8, and TTF-1) to establish the cell lineage and to assess the differentiation status of each of the cell lines. We screened for mutations in the most relevant oncogenes/tumor suppressor genes affected in thyroid carcinogenesis: RAS, BRAF, CTNNB1, and TP53 along with RET/PTC rearrangements. Considering the putative relevance in general carcinogenesis, we have also studied other molecules such as EGFR, PI3K, RAF-1, and THRB. To determine the genetic identity of the cell lines, we performed genotypic analysis. MAIN OUTCOME: The panel of cell lines we have studied displayed activation of several oncogenes (BRAF, RAS, RET/PTC) and inactivation of tumor suppressor genes (TP53) known to be important for thyroid carcinogenesis. Two of the cell lines-TPC1 and FB2-shared the same genotypic profile, probably representing clones of an ancestor cell line (TPC1). CONCLUSION: Due to their different molecular alterations, these cell lines represent a valuable tool to study the molecular mechanisms underlying thyroid carcinogenesis. We suggest that genotypic analyses should be included as a routine procedure to guarantee the uniqueness of each cell line used in research.
Assuntos
Adenocarcinoma Folicular/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Neoplasias da Glândula Tireoide/genética , Adenocarcinoma Folicular/patologia , Adenocarcinoma Papilar/genética , Adenocarcinoma Papilar/patologia , Diferenciação Celular/genética , Linhagem Celular Tumoral , Análise Mutacional de DNA , Genótipo , Humanos , Repetições de Microssatélites , Neoplasias da Glândula Tireoide/patologiaRESUMO
A high prevalence of the BRAF(V600E) somatic mutation was recently reported in several series of papillary thyroid carcinomas (PTC). This mutation appears to be particularly prevalent in PTC with a predominantly papillary architecture. Another BRAF mutation (K601E) was detected in a follicular adenoma and in some cases of the follicular variant of PTC. The few studies on record provided controversial data on the relationship between the occurrence of BRAF mutations and clinicopathologic parameters such as gender, age and tumour staging. In an attempt to clarify such controversies we decided to enlarge our previous series to 315 tumours or tumour-like lesions diagnosed in 280 patients, including a thorough analysis of several clinicopathologic features. The BRAF(V600E) mutation was exclusively detected in PTC with a papillary or mixed follicular/papillary architecture both of the conventional type (46%) and of other histotypes, such as microcarcinoma (43%), Warthin-like PTC (75%) and oncocytic variant of PTC (55%). The BRAF(K601E) mutation was detected in four of the 54 cases of the follicular variant of PTC (7%). The mean age of patients with conventional PTC harbouring BRAF(V600E) (46.7 years) was significantly higher (P<0.0001) than that of patients with conventional PTC without BRAF(V600E) (29.5 years). The BRAF (BRAF(V600E)) mutated PTC did not exhibit signs of higher aggressiveness (size, vascular invasion, extra-thyroid extension and nodal metastasis) and were in fact less often multicentric than PTC without the mutation.
Assuntos
Carcinoma Papilar/genética , Proteínas Proto-Oncogênicas B-raf/genética , Neoplasias da Glândula Tireoide/genética , Adenocarcinoma/genética , Adulto , Fatores Etários , Carcinoma Papilar, Variante Folicular/genética , Análise Mutacional de DNA , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples , Fatores SexuaisRESUMO
Members of a family of thyroid cell lines (KAT) were analyzed because they expressed a higher molecular weight (135 kd) form of E-cadherin at their surface. We found that this aberrant E-cadherin is the result of a point mutation in the exon 9 donor splice site causing a skipping of exon 9 with consequent deletion of the corresponding aminoacids on E-cadherin protein. As a spin-off, we report that the various members of the KAT family share this mutation as well as the genetic background. Furthermore we found that this mutated protein leads to disturbed cell-cell adhesion although E-cadherin is still able to mediate the formation of the cadherin/ catenin complex. We also demonstrate the presence of another cell-cell adhesion complex, formed by Pcadherin and the catenins. The latter is also not able to mediate cell-cell adhesion. Although these cells lack cell-cell adhesion they are not invasive without exogenous stimulus.
Assuntos
Caderinas/genética , Caderinas/metabolismo , Carcinoma/fisiopatologia , Genoma Humano , Mutação , Neoplasias da Glândula Tireoide/fisiopatologia , Sequência de Bases , Carcinoma/genética , Carcinoma/metabolismo , Carcinoma/patologia , Adesão Celular , Linhagem Celular Tumoral , Proteínas do Citoesqueleto/metabolismo , Éxons , Humanos , Invasividade Neoplásica , Mutação Puntual , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/patologia , Transativadores/metabolismo , alfa Catenina , beta CateninaRESUMO
BACKGROUND: Diabetes, a highly prevalent, chronic disease, is associated with increasing frailty and functional decline in older people, with concomitant personal, social, and public health implications. We describe the rationale and methods of the multi-modal intervention in diabetes in frailty (MID-Frail) study. METHODS/DESIGN: The MID-Frail study is an open, randomised, multicentre study, with random allocation by clusters (each trial site) to a usual care group or an intervention group. A total of 1,718 subjects will be randomised with each site enrolling on average 14 or 15 subjects. The primary objective of the study is to evaluate, in comparison with usual clinical practice, the effectiveness of a multi-modal intervention (specific clinical targets, education, diet, and resistance training exercise) in frail and pre-frail subjects aged ≥70 years with type 2 diabetes in terms of the difference in function 2 years post-randomisation. Difference in function will be measured by changes in a summary ordinal score on the short physical performance battery (SPPB) of at least one point. Secondary outcomes include daily activities, economic evaluation, and quality of life. DISCUSSION: The MID-Frail study will provide evidence on the clinical, functional, social, and economic impact of a multi-modal approach in frail and pre-frail older people with type 2 diabetes. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01654341.
Assuntos
Diabetes Mellitus Tipo 2/terapia , Dieta , Conhecimentos, Atitudes e Prática em Saúde , Educação de Pacientes como Assunto , Projetos de Pesquisa , Treinamento Resistido , Atividades Cotidianas , Fatores Etários , Idoso , Protocolos Clínicos , Terapia Combinada , Análise Custo-Benefício , Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Mellitus Tipo 2/economia , Dieta/efeitos adversos , Dieta/economia , Europa (Continente) , Idoso Fragilizado , Custos de Cuidados de Saúde , Humanos , Educação de Pacientes como Assunto/economia , Qualidade de Vida , Treinamento Resistido/economia , Fatores de Tempo , Resultado do TratamentoRESUMO
Cancer development involves multiple genetic changes, which can occur in tumor suppressor genes and lead to loss of function in a recessive manner. Recent findings have identified a novel tumor suppressor gene named GRIM-19. Similar to what has been observed for other known tumor suppressor proteins such as p53, GRIM-19 gene mutations and loss of protein expression have been observed in several tumor types. In this review, we perform a detailed description on the current understanding of GRIM-19 function in carcinogenesis.
Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Transformação Celular Neoplásica/metabolismo , Proteínas Mitocondriais/metabolismo , NADH NADPH Oxirredutases/metabolismo , Neoplasias/metabolismo , Animais , Proteínas Reguladoras de Apoptose/genética , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Transporte de Elétrons , Expressão Gênica , Fator Estimulador de Colônias de Granulócitos/metabolismo , Humanos , Proteínas Mitocondriais/genética , NADH NADPH Oxirredutases/genética , Neoplasias/patologia , Fator de Transcrição STAT3/metabolismoRESUMO
The function of immune cells is critically dependent on their capacity to respond to a complex series of navigational cues that enable them to home to various organ sites in the body or to respond to inflammatory cues such as those released at sites of tissue damage. From early embryonic stages, immune cells are faced with a barrage of signals that will not all be directing the cell to do the same thing. Here we use the Drosophila embryo to investigate how hemocytes (Drosophila macrophages), are able to prioritize key guidance signals and ignore others so that they are not pulled every which way. We identify the immediate wound attractant signal as H(2)O(2) and investigate how Drosophila macrophages respond to competing guidance cues-those emanating from a wound-versus standard developmental guidance cues, as well as those signals drawing cells toward neighboring dying cells. We reveal a hierarchy of responsiveness to attractant cues that varies over time and we identify why there is a wound refractile period early in embryonic development when macrophages cannot be distracted from their developmental migratory pathway to a site of tissue damage.
Assuntos
Drosophila/embriologia , Drosophila/crescimento & desenvolvimento , Macrófagos/citologia , Macrófagos/fisiologia , Transdução de Sinais/fisiologia , Animais , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Hemócitos/citologia , Hemócitos/fisiologia , Peróxido de Hidrogênio/metabolismo , Ferimentos e LesõesRESUMO
Drosophila melanogaster macrophages are highly migratory cells that lend themselves beautifully to high resolution in vivo imaging experiments. By expressing fluorescent probes to reveal actin and microtubules, we can observe the dynamic interplay of these two cytoskeletal networks as macrophages migrate and interact with one another within a living organism. We show that before an episode of persistent motility, whether responding to developmental guidance or wound cues, macrophages assemble a polarized array of microtubules that bundle into a compass-like arm that appears to anticipate the direction of migration. Whenever cells collide with one another, their microtubule arms transiently align just before cell-cell repulsion, and we show that forcing depolymerization of microtubules by expression of Spastin leads to their defective polarity and failure to contact inhibit from one another. The same is true in orbit/clasp mutants, indicating a pivotal role for this microtubule-binding protein in the assembly and/or functioning of the microtubule arm during polarized migration and contact repulsion.