RESUMEN
Ocular albinism type 1 (OA1) is an X-linked disorder characterized by severe impairment of visual acuity, retinal hypopigmentation and the presence of macromelanosomes. We isolated a novel transcript from the OA1 critical region in Xp22.3-22.2 which is expressed at high levels in RNA samples from retina, including the retinal pigment epithelium, and from melanoma. This gene encodes a protein of 424 amino acids displaying several putative transmembrane domains and sharing no similarities with previously identified molecules. Five intragenic deletions and a 2 bp insertion resulting in a premature stop codon were identified from DNA analysis of patients with OA1, indicating that we have identified the OA1 gene.
Asunto(s)
Albinismo Ocular/genética , Proteínas del Ojo/genética , Glicoproteínas de Membrana , Proteínas de la Membrana/genética , Cromosoma X , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Codón de Terminación , ADN Complementario , Femenino , Mutación del Sistema de Lectura , Humanos , Masculino , Melanoma/metabolismo , Datos de Secuencia Molecular , ARN Mensajero/análisis , Mapeo Restrictivo , Retina/metabolismo , Eliminación de SecuenciaRESUMEN
OBJECTIVE: During pancreatic surgery for malignancies, hepatic revascularization is needed in case of en bloc resection with hepatic artery involvement. In these cases, the use of the splenic artery is described in the literature, including transposition and interposition techniques. PATIENTS AND METHODS: We report the case of pancreatic cancer resection with involvement of the right hepatic artery, anomalous arising from the superior mesenteric artery, and hepatic revascularization with splenic artery reconstruction. A literature review to analyze the use of splenic artery in hepatic revascularization during pancreatic cancer surgery was performed. RESULTS: A 61-year-old man with a 55-mm hypovascular tumor in the pancreatic head, in wide contact with the right hepatic artery, underwent total pancreatectomy and splenectomy. Right hepatic artery was resected, and the distal part of the splenic artery was transposed to the right hepatic artery with a termino-terminal anastomosis. Histopathological examination revealed R0 resection. CONCLUSIONS: Hepatic revascularization with splenic artery should be considered in patients suitable to extend resectability in pancreatic cancer surgery. A multidisciplinary approach and careful pre-operative planning are essential.
Asunto(s)
Arteria Hepática/cirugía , Hígado/irrigación sanguínea , Hígado/cirugía , Neoplasias Pancreáticas/irrigación sanguínea , Neoplasias Pancreáticas/cirugía , Arteria Esplénica/cirugía , Humanos , Masculino , Persona de Mediana Edad , Procedimientos de Cirugía Plástica , Procedimientos Quirúrgicos VascularesRESUMEN
OBJECTIVE: This study aims to analyze the early and late outcomes of our 30-year experience with mycotic aneurysms of the abdominal aorta and iliac arteries. PATIENTS AND METHODS: This retrospective cohort study compared the outcomes of all the patients with mycotic aneurysm, by analyzing prospectively collected data between September 1989 and October 2019 from the Unit of Vascular Surgery of Fondazione Policlinico Universitario Gemelli - IRCCS in Rome, Italy. RESULTS: Twenty-three patients with mycotic aneurysm were included. Twenty-two patients underwent surgery; one patient arrived at the emergency room with unstable clinical conditions and died before being treated. Fourteen cases (60.9%) were located at the infrarenal aorta, while three cases (13.0%) were pararenal aortic aneurysms. Six cases (26.1%) had an iliac arteries localization. Seventeen patients (77.3%) underwent open surgical repair aneurysmectomy with in situ reconstruction, while three cases (13.6%) underwent extra-anatomic revascularization. Three patients (13.6%) underwent the placement of an endoprosthesis, of whom two underwent hybrid procedures, and one EVAR. The latter underwent an early conversion to open repair due to a type I endoleak. The mean length of hospital stay was 35 ± 18.7 days. Five patients (22.7%) died in the immediate postoperative period. In the follow-up of 45.5 ± 41.3 months (range 2-156), we documented six deaths (35.3%), of whom two (11.8%) were aortic-related for a 34.8% overall aortic-related mortality. Eleven patients were alive, with an overall survival of 47.8%. CONCLUSIONS: Mycotic aneurysm is an extremely rare and varied pathology. Open surgical repair showed to be a safe approach because of a complete and aggressive debridement of local infected tissues, with an acceptable long-term mortality rate.
Asunto(s)
Aneurisma Infectado/cirugía , Aneurisma de la Aorta Abdominal/cirugía , Aneurisma Ilíaco/cirugía , Arteria Ilíaca/cirugía , Anciano , Anciano de 80 o más Años , Aneurisma Infectado/diagnóstico , Aneurisma de la Aorta Abdominal/diagnóstico , Femenino , Estudios de Seguimiento , Humanos , Aneurisma Ilíaco/diagnóstico , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Tomografía Computarizada por Rayos XRESUMEN
We show that human osteosarcoma cells (Saos-2) have downregulation of alpha3beta1-integrin compared to normal bone cells; this was further described in human osteosarcomas and in a primary murine sarcoma. The alpha3 gene was silenced in Saos-2 cells causing a low expression of alpha3beta1-integrin and reduction in collagen attachment with increasing migratory capacity. Chromatin immunoprecipitation assay performed on alpha3 promoter established that Myc and Yin Yang protein (YY1) cooperate in tandem to downregulate the alpha3 gene. This silencing mechanism involves the binding of Myc and YY1 to DNA and formation of complexes among Myc/Max, YY1, CREB-binding protein and deacetylation activity. The promoter containing deletions of E-boxes or YY1 cassettes failed to downregulate the transcription of a reporter gene as well as the inhibition of deacetylation activity. Overexpression of both Myc and YY1 was necessary to determine the alpha3-integrin promoter downregulation in normal osteoblasts. This downregulation of alpha3beta1-integrin can contribute to the acquisition of a more aggressive phenotype. YY1 regulated negatively the Myc activity through a direct interaction with the Myc/Max and deacetylase complexes. This represents a novel silencing mechanism with broad implications in the transcription machinery of tumours.
Asunto(s)
Silenciador del Gen , Integrina alfa3/genética , Integrina alfa3beta1/metabolismo , Osteosarcoma/metabolismo , Proteínas Proto-Oncogénicas c-myc/metabolismo , Factor de Transcripción YY1/metabolismo , Animales , Secuencia de Bases , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Western Blotting , Neoplasias Óseas/metabolismo , Neoplasias Óseas/patología , Inmunoprecipitación de Cromatina , Colágeno/metabolismo , Regulación hacia Abajo , Ensayo de Cambio de Movilidad Electroforética , Femenino , Técnica del Anticuerpo Fluorescente , Humanos , Laminina/metabolismo , Ratones , Ratones Desnudos , Datos de Secuencia Molecular , Osteoblastos/citología , Osteoblastos/metabolismo , Osteosarcoma/genética , Osteosarcoma/patología , Regiones Promotoras Genéticas/genética , ARN Interferente Pequeño/farmacología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transcripción Genética , Transfección , Células Tumorales Cultivadas , Factor de Transcripción YY1/antagonistas & inhibidores , Factor de Transcripción YY1/genéticaRESUMEN
OBJECTIVE: To evaluate the role of quantitative digital subtraction angiography (Q-DSA) with parametric color coding (PCC) in assessing patients with type B chronic thoracic aortic dissection (TBCAD) during thoracic endovascular aortic repair (TEVAR) procedures. PATIENTS AND METHODS: A total of 11 patients electively treated in our Department for a TBCAD were retrospectively enrolled. All cases were treated with TEVAR for false lumen aneurysm of the thoracic descending aorta. For digital subtraction angiography (DSA) series post-processing, a newly implemented PCC algorithm was used to turn consecutive two-dimensional images into a single color-coded picture (syngo iFLOW, Siemens AG, Forchheim, Germany). In consensus reading, two clinicians experienced in vascular imaging evaluated the DSA series in blinded assessment and compared them to the color-coded images. PCC was assessed for its accuracy in identifying the true and false lumen as well as whether it could provide improved visualization in pre-deployment stent grafting and the final evaluation of treatment. RESULTS: PCC facilitated the visualization of the aortic dissection angioarchitecture in terms of contemporary true and false lumen vision in 81.8% of the cases. In 72.7% of the procedures, Q-DSA was estimated to improve aorta information assessment in terms of false lumen viewing, and it was possible to identify the proximal entry tear position in 45.4% of the cases. After stent graft deployment, in 72.7% of the cases (all 8 patients in which the aortic arch false lumen was visible in pre-treatment), Q-DSA confirmed the absence of early false lumen reperfusion. CONCLUSIONS: Our results indicate that Q-DSA could be useful in the intraprocedural evaluation of patients with aortic dissection during TEVAR procedures without additional x-ray costs and contrast exposure.
Asunto(s)
Angiografía de Substracción Digital , Aneurisma de la Aorta Torácica/patología , Disección Aórtica/patología , Anciano , Algoritmos , Disección Aórtica/diagnóstico por imagen , Disección Aórtica/cirugía , Aorta/diagnóstico por imagen , Aneurisma de la Aorta Torácica/diagnóstico por imagen , Aneurisma de la Aorta Torácica/cirugía , Procedimientos Endovasculares , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador , Masculino , Estudios Retrospectivos , Stents , Tomografía Computarizada por Rayos X , Resultado del TratamientoRESUMEN
OBJECTIVE: Maternal hypercholesterolaemia during pregnancy increases lipid peroxidation in mothers and fetuses and programs increased susceptibility to atherosclerosis later in life. The objective of this study was to elucidate the role of the placenta in mediating oxidative stress from mother to offspring. DESIGN: Comparison between normo- and hypercholesterolaemic mothers (n = 36 each) and their children. SETTING: Obstetric wards, hospitals of the University of Naples and Regione Campania. POPULATION: Healthy primiparas delivering by caesarean section. METHODS: Biochemical measurements of oxidative stress and serum leptin in cord plasma and placenta, immunochemistry of placenta microvessels, and vasoreactivity studies were performed. MAIN OUTCOME MEASURES: Oxidative status (i.e. lipid composition and content of oxidised fatty acids, activity of pro- and antioxidant enzymes, immunohistochemical presence of oxidation-specific epitopes) in maternal and cord blood and in placental tissue, as well as vascular reactivity in omental arteries. RESULTS: Hypercholesterolaemia during pregnancy was associated with extensive changes in fatty acid composition of both maternal and cord blood lipids, sufficient to alter vasoreactivity of omental vessels. Results also indicated that the placenta is not only subject to substantial oxidative stress, but that it may further increase fetal oxidative stress through changes of pro- and antioxidant enzyme activities. CONCLUSIONS: The placenta plays an important role in both transmitting and enhancing pathogenic effects of gestational hypercholesterolaemia.
Asunto(s)
Ácidos Grasos/química , Hipercolesterolemia/metabolismo , Epiplón/irrigación sanguínea , Placenta/enzimología , Complicaciones del Embarazo/metabolismo , Adulto , Arterias/fisiología , Ácidos Grasos/administración & dosificación , Femenino , Sangre Fetal/química , Edad Gestacional , Humanos , Inmunohistoquímica , Leptina/metabolismo , Peroxidación de Lípido/fisiología , Lípidos/sangre , Lípidos/química , Contracción Muscular/efectos de los fármacos , Músculo Liso Vascular/efectos de los fármacos , Oxidación-Reducción , Estrés Oxidativo/fisiología , Embarazo , Vasoconstrictores/farmacología , Sistema Vasomotor/metabolismoRESUMEN
Different rehabilitation models for persons diagnosed with disorders of consciousness have been proposed in Europe during the last decade. In Italy, the Ministry of Health has defined a national healthcare model, although, to date, there is a lack of information on how this has been implemented at regional level. The INCARICO project collected information on different regional regulations, analysing ethical aspects and mapping care facilities (numbers of beds and medical units) in eleven regional territories. The researchers found a total of 106 laws; differences emerged both between regions and versus the national model, showing that patients with the same diagnosis may follow different pathways of care. An ongoing cultural shift from a treatment-oriented medical approach towards a care-oriented integrated biopsychosocial approach was found in all the welfare and healthcare systems analysed. Future studies are needed to explore the relationship between healthcare systems and the quality of services provided.
Asunto(s)
Necesidades y Demandas de Servicios de Salud , Estado Vegetativo Persistente/rehabilitación , Política de Salud , Capacidad de Camas en Hospitales , Humanos , Italia , Programas Nacionales de Salud , RegionalizaciónRESUMEN
Maternal hypercholesterolemia during pregnancy is associated with a marked increase in aortic fatty streak formation in human fetuses and faster progression of atherosclerosis during normocholesterolemic childhood. However, the mechanisms responsible are unknown, and the contribution of genetic differences is difficult to assess in humans. The goal of this study was to determine whether maternal hypercholesterolemia per se may cause enhanced fatty streak formation in offspring and whether interventions during pregnancy can reduce it. During pregnancy, 1 group of New Zealand White rabbits was fed control chow and 8 groups were fed hypercholesterolemic diets Chol 1 (yielding plasma cholesterol of 153 mg/dL) or Chol 2 (yielding 359 mg/dL) without or with cholestyramine, vitamin E, or both. Offspring (n=15 to 25 per group) were killed at birth. Maternal hypercholesterolemia enhanced mean lesion size in the aorta of their offspring at birth from 44+/-18x10(3) micrometer(2) per section in controls to 85+/-26x10(3) in Chol 1 and 156+/-49x10(3) in Chol 2 groups (P<0.0001 for both). Cholestyramine or vitamin E treatment of mothers significantly reduced atherosclerosis at birth by up to 39% compared with controls on the same diet. Oxidized fatty acids and malondialdehyde in aortic atherosclerotic lesions and plasma were similarly affected by diets and treatment as atherosclerosis. Our results establish the causal role of hypercholesterolemia and peroxidation in fetal atherogenesis and demonstrate that both lipid-lowering and antioxidant interventions during pregnancy can reduce it. If it can be established that interventions in mothers also affect progression of lesions after birth, this may indicate a novel approach for the prevention of atherosclerosis.
Asunto(s)
Arteriosclerosis/congénito , Arteriosclerosis/prevención & control , Resina de Colestiramina/administración & dosificación , Hipercolesterolemia/tratamiento farmacológico , Complicaciones del Embarazo/tratamiento farmacológico , Efectos Tardíos de la Exposición Prenatal , Vitamina E/administración & dosificación , Animales , Antioxidantes/administración & dosificación , Aorta/efectos de los fármacos , Aorta/patología , Arteriosclerosis/sangre , Arteriosclerosis/patología , Colesterol/sangre , Colesterol en la Dieta/farmacología , Dieta Aterogénica , Ácidos Grasos/metabolismo , Femenino , Feto/efectos de los fármacos , Feto/metabolismo , Feto/patología , Hipercolesterolemia/sangre , Peroxidación de Lípido/efectos de los fármacos , Embarazo , Complicaciones del Embarazo/sangre , Conejos , Inducción de Remisión , Triglicéridos/sangreRESUMEN
Maternal hypercholesterolemia during pregnancy is associated with enhanced fatty streak formation in human fetuses and faster progression of atherosclerosis during childhood even under normocholesterolemic conditions. A causal role of maternal hypercholesterolemia in lesion formation during fetal development has previously been established in rabbits. The same experimental model is now used to establish that maternal hypercholesterolemia or ensuing pathogenic events in fetal arteries enhance atherogenesis later in life. Five groups of rabbit mothers were fed chow, cholesterol-enriched chow, or cholesterol-enriched chow plus 1000 IU vitamin E, 3% cholestyramine, or both during pregnancy. Offspring of all groups (n=136) were fed a mildly hypercholesterolemic diet for up to a year and had similar cholesterol levels. Aortic lesion sizes and lipid peroxidation products in plasma and lesions in offspring were determined at birth, 6 months, or 12 months. Lesion progression in offspring of hypercholesterolemic mothers was greater than in all other groups. At each time point, offspring of hypercholesterolemic mothers had 1.5- to 3-fold larger lesions than offspring of normocholesterolemic mothers (P<0.01), with the greatest absolute differences at 12 months. Maternal treatment reduced lesions by 19% to 53%, compared with offspring of untreated hypercholesterolemic mothers (P<0.01), with the greatest effect in the vitamin E groups. At 12 months, lesions in offspring of all vitamin E and cholestyramine-treated mothers were similar to those of normocholesterolemic mothers. Lipid peroxidation end-products in lesions and plasma showed analogous differences between groups as lesions (P<0.01). Thus, pathogenic programming in utero increases the susceptibility to atherogenic risk factors later in life and maternal intervention with cholesterol-lowering drugs or antioxidants reduce postnatal lipid peroxidation and atherosclerosis in their offspring.
Asunto(s)
Arteriosclerosis/etiología , Hipercolesterolemia/tratamiento farmacológico , Animales , Anticolesterolemiantes/uso terapéutico , Antioxidantes/uso terapéutico , Aorta/patología , Enfermedades de la Aorta/sangre , Enfermedades de la Aorta/etiología , Enfermedades de la Aorta/patología , Arteriosclerosis/sangre , Arteriosclerosis/patología , Resina de Colestiramina/uso terapéutico , Progresión de la Enfermedad , Femenino , Ácido Linoleico/sangre , Peroxidación de Lípido , Malondialdehído/sangre , Embarazo , Conejos , Vitamina E/uso terapéuticoRESUMEN
A subtractive library screening was performed to identify changes in gene expression that occur during the process of neoplastic transformation of thyroid cells. A cDNA library was constructed from a human thyroid papillary carcinoma cell line (NPA) subtracted with cDNAs from normal thyroid cells (HTC 2). The differential screening of this library lead to the isolation of 39 cDNA clones; six of them showed homology with a recently isolated gene, named HIP, that codes for a protein belonging to a novel class of heparin/heparan sulfate-binding proteins. Northern blot analysis revealed HIP gene overexpression in all of the human thyroid carcinoma cell lines analyzed, as compared to the HTC 2 cells. HIP expression was particularly abundant in the anaplastic carcinoma-derived cell lines. The analysis of surgically removed thyroid tumors showed overexpression of HIP gene in all of the carcinomas, independent of the histotype, although the largest increase in HIP expression was observed in the undifferentiated forms. In contrast, none of the benign adenomas or normal thyroid tissues showed HIP overexpression. To establish the role of HIP overexpression in cell transformation, the NPA cell line was transfected with an eukaryotic expression vector carrying the HIP gene in the antisense orientation. Stable transfectants expressed reduced HIP mRNA levels and showed morphological changes, such as becoming spindle-shaped and growing scattered. The growth rate of the antisense clones was greatly reduced compared to the NPA cells transfected with the backbone vector. Taken together, these results indicate that HIP gene overexpression is associated with thyroid carcinogenesis and strongly suggest its involvement in thyroid cell growth regulation.
Asunto(s)
Carcinoma Papilar/metabolismo , Proteínas Portadoras/genética , Regulación Neoplásica de la Expresión Génica , Heparina/metabolismo , Heparitina Sulfato/metabolismo , Neoplasias de la Tiroides/metabolismo , Humanos , Glándula Tiroides/metabolismo , Células Tumorales Cultivadas , Regulación hacia ArribaRESUMEN
The proteins of the Ets family are transcription factors involved in signal transduction, cell cycle progression, and differentiation. In this study, we report that thyroid cell neoplastic transformation is associated with a dramatic increase in ETS transcriptional activity, which is dependent on the accumulation of Ets-1, Ets-2, and other Ets-related proteins. Inhibition of ETS transactivation activity by the Ets-dominant negative construct (Ets-Z) induced programmed cell death in human thyroid carcinoma cell lines but not in normal thyroid cells. Apoptotic cell death induced by Ets-Z was dependent on the reduction of c-MYC protein levels, because it was prevented by overexpression of c-myc. Taken together, these data indicate that the induction of Ets-1 and Ets-2 transcription factors plays a pivotal role in thyroid cell neoplastic transformation.
Asunto(s)
Transformación Celular Neoplásica/metabolismo , Proteínas de Unión al ADN , Proteínas Proto-Oncogénicas/biosíntesis , Proteínas Represoras , Glándula Tiroides/metabolismo , Neoplasias de la Tiroides/metabolismo , Transactivadores/biosíntesis , Factores de Transcripción/biosíntesis , Apoptosis/fisiología , Secuencia de Consenso , ADN/metabolismo , Genes myc/genética , Humanos , Proteína Proto-Oncogénica c-ets-1 , Proteína Proto-Oncogénica c-ets-2 , Proteínas Proto-Oncogénicas/fisiología , Proteínas Proto-Oncogénicas c-ets , Glándula Tiroides/patología , Neoplasias de la Tiroides/patología , Transactivadores/fisiología , Factores de Transcripción/fisiología , Activación Transcripcional/fisiología , Células Tumorales CultivadasRESUMEN
Expression of mutated versions of the p53 gene deranged the differentiation program of thyroid cells and resulted in deregulated growth. Specifically, p53 mutants in several residues of the DNA-binding region induced thyrotropin (TSH) -independent growth and inhibition of the expression of thyroid-specific genes. The loss of the differentiated phenotype invariably correlated with the blockage of the expression of the genes coding for the thyroid transcriptional factors PAX-8 and TTF2. Conversely, thyroid cells transfected with a p53 gene mutated at codon 392, located outside the DNA-binding region, stimulated the expression of differentiation genes in the absence of the TSH, and induced TSH-independent growth. cAMP intracellular levels were higher in thyroid cells transfected with the p53 gene mutated at the 392 site than in the untransfected thyroid cells, but lower in the cells transfected with the other mutated p53 genes. Fra-1 and c-jun were induced by p53, resulting in increased AP-1 levels. The results of this study suggest that p53 exerts effects on cAMP transduction pathway in thyroid cells, which are exquisitely sensitive to cAMP.
Asunto(s)
Diferenciación Celular/genética , Genes p53/fisiología , Glándula Tiroides/citología , Animales , Sitios de Unión , División Celular/genética , Células Cultivadas , AMP Cíclico/metabolismo , Genes p53/genética , Mutación , Peroxidasas/genética , Peroxidasas/metabolismo , Fenotipo , Ratas , Receptores de Tirotropina/genética , Receptores de Tirotropina/metabolismo , Tiroglobulina/genética , Tiroglobulina/metabolismo , Factor de Transcripción AP-1/metabolismo , Transfección , Proteína p53 Supresora de Tumor/química , Proteína p53 Supresora de Tumor/genéticaRESUMEN
Hyperplastic or neoplastic proliferative lesions of thyroid follicular epithelium consist of a spectrum, ranging from nodular hyperplasia to undifferentiated (anaplastic) carcinoma, and usually present as palpable thyroid nodules. Thyroid nodules are a common occurrence in the general population, but only a small proportion of them are eventually diagnosed as carcinoma. The difficulty in objectively identifying those thyroid nodules that are malignant to avoid unnecessary surgery, combined with the range and effectiveness of the available therapeutic options in those patients who do, indeed, have thyroid carcinoma, has prompted the search for tumor markers and prognostic indicators. The high mobility group I (HMGI) proteins represent a class of nuclear proteins involved in the regulation of chromatin structure and function. HMGI(Y), one of the members of this class, is expressed at high levels during embryogenesis and in malignant tumors but at generally low levels in normal adult human tissues. Previous work on a limited number of thyroid samples suggested that the detection of the HMGI(Y) proteins may provide a clinically useful diagnostic tool. To verify this assumption, we analyzed HMGI(Y) expression by a combination of immunohistochemistry and reverse transcription-PCR in 358 thyroid tissue samples that were representative of the spectrum of thyroid tumor pathology. HMGI(Y) was detectable in 18 of 19 follicular carcinomas, 92 of 96 papillary carcinomas, and 11 of 11 undifferentiated (anaplastic) carcinomas but in only 1 of 20 hyperplastic nodules, 44 of 200 follicular adenomas, and 0 of 12 normal tissue samples. The correlation between HMGI(Y) expression and a diagnosis of carcinoma was highly significant (P < 0.0001). We also prospectively collected and analyzed for HMGI(Y) expression by immunohistochemistry and reverse transcription-PCR in 12 fine needle aspiration biopsies from 10 patients who subsequently underwent surgical removal of a solitary thyroid nodule. HMGI(Y) was detectable only in the four fine needle aspiration biopsies, corresponding to the thyroid nodules that were definitively diagnosed as carcinomas after surgery (two follicular carcinomas and two papillary carcinomas). The remaining eight samples (six follicular adenomas and two samples consisting of normal follicular cells) were negative. The findings of this study confirm the differential expression of HMGI(Y) in thyroid neoplasia and indicate the HMGI(Y) protein as a potential marker for thyroid carcinoma.
Asunto(s)
Adenocarcinoma Folicular/química , Adenoma/química , Carcinoma/química , Proteínas del Grupo de Alta Movilidad/análisis , Proteínas de Neoplasias/análisis , Neoplasias de la Tiroides/química , Adenocarcinoma Folicular/diagnóstico , Adenoma/diagnóstico , Adulto , Biopsia con Aguja , Carcinoma/diagnóstico , Carcinoma Papilar/química , Carcinoma Papilar/diagnóstico , Regulación de la Expresión Génica , Humanos , Inmunohistoquímica , Reacción en Cadena de la Polimerasa , Estudios Prospectivos , Neoplasias de la Tiroides/diagnósticoRESUMEN
We have investigated the role of the NFkappaB complex in the process of thyroid carcinogenesis by analysing thyroid carcinoma cell lines. A significant increase in p65 NFkappaB mRNA and protein expression, compared to normal thyroid cultures or tissue, was found in all of the cancer cell lines. Conversely, only a modest increase in the p50 NFkappaB mRNA and protein was found in most, but not all carcinoma cell lines. The block of p65 protein synthesis with specific antisense oligonucleotides greatly reduced the ability of two undifferentiated carcinoma cell lines to form colonies in agar and reduced their growth rate. On the other hand, no effect was observed in the same cell lines when treated with p50 specific antisense oligonucleotides. These inhibitory effects seem to be mediated by the suppression of c-myc gene expression, since treatment with antisense oligonucleotides for p65 gene interfered negatively with c-myc gene expression. Our results indicate that activation of the NFkappaB complex by overexpression of p65 plays a critical role in the process of thyroid cell transformation.
Asunto(s)
FN-kappa B/genética , Neoplasias de la Tiroides/genética , Regulación hacia Abajo/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica , Genes myc , Humanos , FN-kappa B/biosíntesis , Oligonucleótidos Antisentido/farmacología , Fenotipo , Inhibidores de la Síntesis de la Proteína/farmacología , Neoplasias de la Tiroides/metabolismo , Células Tumorales CultivadasRESUMEN
We have recently reported that neoplastic transformation of two rat thyroid epithelial cell lines by retroviruses carrying the v-mos and v-ras Ki oncogenes is associated with a drastic increase of AP-1 activity. The most important effects were represented by the dramatic junB and fra-1 gene induction, which was abolished by the block of the transformation-induced HMGI-C protein synthesis. Here, we have further characterized the transformation-dependent AP-1 activity, by analysing the expression of different jun- and fos-related components, in rat thyroid cell lines transformed by several oncogenes, in human thyroid carcinoma cell lines, and in naturally occurring human thyroid tumours. A significant increase of Fra-1 and JunB protein levels was detected in all oncogene transformed rat thyroid cell lines. Fra-1 gene induction was demonstrated to occur also in human thyroid carcinoma cell lines and tissues. Conversely, c-Jun and JunD proteins, rather than JunB, accumulated in human thyroid carcinoma cell lines. An induction of AP-1 target genes was also detected both in rat and human thyroid transformed cell lines. Therefore, in vivo and in vitro thyroid cell transformation is associated with important compositional changes in the AP-1 complex and an increased transcriptional activity.
Asunto(s)
Transformación Celular Neoplásica , Oncogenes , Glándula Tiroides/metabolismo , Neoplasias de la Tiroides/genética , Factor de Transcripción AP-1/metabolismo , Animales , Células Epiteliales/metabolismo , Células Epiteliales/patología , Genes jun , Proteína HMGA2 , Proteínas del Grupo de Alta Movilidad/biosíntesis , Humanos , Proteínas de Neoplasias/biosíntesis , Reacción en Cadena de la Polimerasa , Proteínas Proto-Oncogénicas c-fos/biosíntesis , Proteínas Proto-Oncogénicas c-jun/biosíntesis , Ratas , Tiroglobulina/biosíntesis , Glándula Tiroides/patología , Tirotropina/farmacologíaRESUMEN
BACKGROUND: Atherosclerotic lesions in intracranial arteries occur later and are less extensive than in extracranial arteries. To investigate potential mechanisms responsible for this difference, in particular the atherogenic response to hypercholesterolemia and LDL oxidation, we compared the extent of fatty streak formation and the composition of these very early lesions in intracranial arteries of human fetuses from normocholesterolemic and hypercholesterolemic mothers with those in extracranial arteries. METHODS AND RESULTS: Lesions were quantified by computer-assisted image analysis of 30 oil red O-stained sections, each from the middle cerebral, basilar, and common carotid arteries and the abdominal aorta of human fetuses (spontaneous abortions and premature newborns who died within 12 hours of birth; both of fetal age 6.2+/-1.3 months) from 43 hypercholesterolemic mothers and 34 normocholesterolemic mothers. Macrophages, apolipoprotein B, and 2 epitopes of oxidized LDL in lesions were determined immunocytochemically. Activities of superoxide dismutase, catalase, and glutathione peroxidase in the arterial wall were also determined. Lesion numbers and sizes were dramatically greater in the abdominal aorta (area of the largest lesion per section: 66.5+/-10.9 x10(3) microm2) and the carotid (11. 6+/-5.3 x10(3) microm2) than in the basilar and middle cerebral artery (0.4+/-0.1 and 0.8+/-0.2 x10(3) microm2, respectively; P<0. 0001). Hypercholesterolemia resulted in a significant increase of lesion size in extracranial arteries but only a marginal increase in intracranial arteries. In analogy, hypercholesterolemia induced a much greater increase in the intimal presence of macrophages, apolipoprotein B, and oxidized LDL (oxidation-specific epitopes) in extracranial than in intracranial arteries. Immunocytochemistry did not indicate that lesions of intracranial arteries contain relatively less oxidized LDL than similar-size lesions of extracranial arteries. Activities of Mn-superoxide dismutase but not of Zn-superoxide dismutase, catalase, or glutathione peroxidase were significantly higher in both intracranial arteries. CONCLUSIONS: Exposure to hypercholesterolemia during fetal development results in extensive formation of fatty streaks in extracranial but not intracranial arteries. The fact that such a difference in lesion formation occurs in the absence of many other atherogenic risk factors found later in life suggests that differences in the atherogenic response to hypercholesterolemia are an important contributor to the slower onset of the disease in intracranial vessels in adults. Fetal arteries may allow elucidation of the mechanisms responsible, for example, better protection of intracranial arteries against free radical-mediated atherogenic processes.
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Aorta Abdominal/embriología , Arteriosclerosis/etiología , Arteria Carótida Común/embriología , Arterias Cerebrales/embriología , Enfermedades Fetales/etiología , Hipercolesterolemia/fisiopatología , Complicaciones del Embarazo/fisiopatología , Aborto Espontáneo , Adulto , Aorta Abdominal/enzimología , Aorta Abdominal/patología , Arteriosclerosis/embriología , Arteriosclerosis/patología , Arteria Carótida Común/enzimología , Arteria Carótida Común/patología , Catalasa/análisis , Arterias Cerebrales/enzimología , Arterias Cerebrales/patología , Femenino , Enfermedades Fetales/patología , Radicales Libres , Edad Gestacional , Glutatión Peroxidasa/análisis , Humanos , Inmunidad Innata , Recién Nacido , Recien Nacido Prematuro , Peroxidación de Lípido , Lípidos/análisis , Masculino , Especificidad de Órganos , Embarazo , Complicaciones del Embarazo/sangre , Superóxido Dismutasa/análisisRESUMEN
BACKGROUND: Oxidized LDL (oxLDL) promotes atherogenesis, and antioxidants reduce lesions in experimental models. OxLDL-mediated effects on c-Myc are poorly characterized, and those on c-Myc nuclear pathways are completely unknown. c-Myc stimulates smooth muscle cell (SMC) proliferation and could be involved in atherosclerosis. We investigated the early effects of oxLDL and alpha-tocopherol on c-Myc, its binding partner Max, and the carboxy-terminal domain-binding factors activator protein-2 and elongation 2 factor in human coronary SMCs. We also investigated whether 9-week treatment of Watanabe heritable hyperlipidemic (WHHL) rabbits with diet-enriched alpha-tocopherol reduces c-Myc expression and oxLDL in the left coronary artery. METHODS AND RESULTS: OxLDL enhanced c-Myc/Max expression and transcription by cotransfection assay and the nuclear activities of E2F and activator protein-2 by binding shift and supershift in coronary SMCs. alpha-Tocopherol significantly reduced these molecular events. Furthermore, alpha-tocopherol reduced early lesions, SMC density, and the immunohistochemical presence of c-Myc, which colocalized with oxLDL/foam cells in the coronaries of WHHL rabbits. CONCLUSIONS: We provide the first evidence that oxLDL and alpha-tocopherol may influence c-Myc activation and several c-Myc-dependent signaling pathways in human coronary SMCs. The observation that in vivo, an antioxidant reduces both c-Myc and oxLDL in early coronary lesions of rabbits is consistent with, but does not prove, the hypothesis that c-Myc-dependent factors activated by oxidative processes contribute to atherogenesis and coronary heart disease.
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Proteínas Portadoras , Proteínas de Ciclo Celular , Hiperlipidemias/metabolismo , Lipoproteínas LDL/farmacología , Músculo Liso Vascular/metabolismo , Proteínas Proto-Oncogénicas c-myc/metabolismo , Transducción de Señal , Vitamina E/farmacología , Animales , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico , Células Cultivadas , Proteínas de Unión al ADN/metabolismo , Factores de Transcripción E2F , Humanos , Hiperlipidemias/tratamiento farmacológico , Factores de Transcripción de Tipo Kruppel , Músculo Liso Vascular/efectos de los fármacos , Oxidación-Reducción , Proteínas Proto-Oncogénicas c-myc/fisiología , Conejos , Proteína 1 de Unión a Retinoblastoma , Factor de Transcripción DP1 , Factores de Transcripción/metabolismo , Vitamina E/uso terapéuticoRESUMEN
New approaches to atherosclerosis-related diseases include novel uses of proven treatments and development of innovative agents. Several commonly used cardiovascular drugs such as dihydropyridine calcium antagonists, ACE inhibitors containing the sulphydryl group, or highly lipophilic beta-blockers have some anti-atherosclerotic activities. Moreover, new clinical trials suggesting that additional reduction of low-density lipoprotein cholesterol levels with statin therapy results in additional benefit in coronary heart disease prevention. Notably, new cholesterol transport or bile acid transport inhibitors have been found to produce significant reductions in intestinal cholesterol absorption and experimental atherosclerosis. Inhibitors of acyl coenzyme A:cholesterol acyltransferase, which can reduce cholesterol storage in macrophages and in arterial lesions, have also been developed. Finally, newer therapeutical strategies against atherogenesis may include the use of antioxidants and cholestyramine during pregnancy or the development of metalloproteinase inhibitors.
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Arteriosclerosis/tratamiento farmacológico , Fármacos Cardiovasculares/uso terapéutico , Fármacos Cardiovasculares/clasificación , Femenino , Humanos , Embarazo , Complicaciones del Embarazo/tratamiento farmacológicoRESUMEN
fra-1 gene overexpression has been shown to represent a general event in thyroid cell transformation in vitro and in vivo. Moreover, inhibition of FRA-1 protein synthesis by stable transfection with a fra-1 antisense construct significantly reduces the malignant phenotype of the transformed thyroid cells, indicating a pivotal role of the fra-1 gene product in the process of cellular transformation. In the attempt to define the potential use of FRA-1 protein detection in the diagnosis of thyroid diseases, we analyzed Fra-1 expression by a combination of immunohistochemistry and reverse transcription-PCR (RT-PCR) assay in 174 samples of thyroid nodules (22 nodular hyperplasias, 102 follicular adenomas, 34 papillary carcinomas, 12 follicular carcinomas, and 4 anaplastic carcinomas) representative of the spectrum of thyroid tumor pathology. FRA-1 protein was abundant in all of the carcinoma samples (50/50, 100%), with an intense staining in the nucleus and the cytoplasm. Positive staining was also found in most of the adenomas (90 of 102; 88%), but in this case, the staining was restricted to the nucleus. Similar results were obtained from the analysis of thyroid goiters; however, the number of positive cases is lower than adenomas (8 of 22; 36%); moreover, the staining was not observed in all of the cells. Conversely, no FRA-1 protein was detectable in 12 normal thyroid tissue samples used as controls. RT-PCR analysis confirmed a higher fra-1 expression in papillary and follicular carcinomas compared with goiters and adenomas. fra-1 expression was also analyzed on 10 fine needle aspiration biopsy (FNAB) samples by RT-PCR. fra-1-specific mRNA was detected in seven of the eight FNABs corresponding to thyroid nodules that were eventually diagnosed as adenomas (three of four) and carcinomas (four of four) after surgery. Conversely, no fra-1 gene expression was observed in two FNABs derived from normal thyroid. Further studies are required before suggesting FRA-1 protein detection as a useful tool for the diagnosis of hyperplastic and neoplastic disorders of the thyroid gland.
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Proteínas Proto-Oncogénicas c-fos/análisis , Proteínas Proto-Oncogénicas c-fos/genética , Glándula Tiroides/patología , Neoplasias de la Tiroides/metabolismo , Nódulo Tiroideo/metabolismo , Biopsia con Aguja , Proteína HMGA1a , Proteínas del Grupo de Alta Movilidad/análisis , Proteínas del Grupo de Alta Movilidad/genética , Humanos , Hiperplasia , Inmunohistoquímica , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias de la Tiroides/patología , Nódulo Tiroideo/patología , Factores de Transcripción/análisis , Factores de Transcripción/genética , Células Tumorales CultivadasRESUMEN
BACKGROUND AND PURPOSE: Atherosclerosis occurs later and is less extensive in intracranial arteries than in extracranial arteries. However, the mechanisms responsible are poorly understood. A previous study has suggested a better antioxidant protection of intracranial arteries. METHODS: To assess the influence of age on arterial activity of antioxidant enzymes and atherogenesis, we compared intracranial and extracranial arteries of humans of different ages who retrospectively lacked confounding classic risk factors (48 premature fetuses aged 6.4+/-0.8 months [mean+/-SD], 58 children aged 7.9+/-3.8 years, 42 adults aged 42.5+/-5.1 years, and 40 elderly subjects aged 71.8+/-3.4 years; all males). Lesions were quantified by computer-assisted imaging analysis of sections of the middle cerebral and basilar arteries, the left anterior descending coronary artery, the common carotid artery, and the abdominal aorta. Macrophages, apolipoprotein B, oxidized LDL, and matrix metalloproteinase-9 in lesions were determined by immunocytochemistry. The effect of aging on atherogenesis was then compared with that on the activity of 4 antioxidant enzymes in the arterial wall. RESULTS: Atherosclerosis was 6- to 19-fold greater (P<0.01) in extracranial arteries than in intracranial arteries, and it increased linearly with age. Intracranial arteries showed significantly greater antioxidant enzyme activities than did extracranial arteries. However, the antioxidant protection of intracranial arteries decreased significantly in older age, coinciding with a marked acceleration of atherogenesis. An increase in matrix metalloproteinase-9 protein expression and in gelatinolytic activity consistent with the degree of intracranial atherosclerosis was also observed. CONCLUSIONS: These results suggest that a greater activity of antioxidant enzymes in intracranial arteries may contribute to their greater resistance to atherogenesis and that with increasing age intracranial arteries respond with accelerated atherogenesis when their antioxidant protection decreases relatively more than that of extracranial arteries.