RESUMO
Hashimoto's thyroiditis (HT) is the most common of all thyroid diseases and is characterized by abundant lymphocyte infiltrate and thyroid impairment, caused by various cell- and antibody-mediated immune processes. Viral infections have been suggested as possible environmental triggers, but conclusive data are not available. We analyzed the presence and transcriptional state of human herpesvirus 6 (HHV-6) in thyroid fine needle aspirates (FNA) and peripheral blood mononuclear cells (PBMCs) from 34 HT patients and 28 controls, showing that HHV-6 DNA prevalence (82% vs. 10%, p≤0.001) and viral load were significantly increased in FNA from HT patients, and thyrocytes from HT FNA displayed a 100-fold higher HHV-6 DNA load compared to infiltrating lymphocytes. In addition, while HHV-6 was strictly latent in positive samples from controls, a low grade acute infection was detected in HT samples. HHV-6 variant characterization was carried out in 10 HT FNA samples, determining that all specimens harbored HHV-6 Variant A.The tropism of HHV-6 for thyroid cells was verified by infection of Nthy-ori3-1, a thyroid follicular epithelial cell line, showing that thyrocytes are permissive to HHV-6 replication, which induces de novo expression of HLA class II antigens. Furthermore, HHV-6-infected Nthy-ori3-1 cells become targets for NK-mediated killing, NK cells from HT patients show a significantly more efficient killing of HHV-6 infected thyroid cells than healthy controls, and HT patients have increased T-cell responses to HHV-6 U94 protein, associated to viral latency. These observations suggest a potential role for HHV-6 (possibly variant A) in the development or triggering of HT.
Assuntos
Doença de Hashimoto/etiologia , Doença de Hashimoto/virologia , Herpesvirus Humano 6/patogenicidade , Infecções por Roseolovirus/virologia , Glândula Tireoide/patologia , Biópsia por Agulha Fina , Linhagem Celular , DNA Viral , Células Epiteliais/virologia , Doença de Hashimoto/imunologia , Herpesvirus Humano 6/genética , Herpesvirus Humano 6/imunologia , Herpesvirus Humano 6/isolamento & purificação , Antígenos de Histocompatibilidade Classe II/biossíntese , Humanos , Células Matadoras Naturais/imunologia , Leucócitos Mononucleares/virologia , Glândula Tireoide/virologia , Carga ViralRESUMO
Multiple endocrine neoplasia (MEN) syndromes are characterized by tumors involving two or more endocrine glands. Two MEN syndromes have long been known: MEN1 and MEN2,caused by germline mutations in MEN1 or RET, respectively. Recently, mutations in CDKN1B,encoding the cyclin-dependent kinase (Cdk) inhibitor p27, were identified in patients having a MEN1-like phenotype but no MEN1 gene mutations. Currently, the molecular mechanisms mediating the role of p27 in tumor predisposition are ill defined. We here report a novel germline missense variant in CDKN1B (c.678C>T, p.P69L) found in a patient with multiple endocrine tumors. We previously reported a nonsense p27 mutation (c.692G>A, p.W76X) in two patients with MEN1-like phenotype. Functional assays were used to characterize p27P69L and p27W76X in vitro. We show that p27P69L is expressed at reduced level and is impaired in both binding toCdk2 and inhibiting cell growth. p27W76X, which is mislocalized to the cytoplasm, can no longer efficiently bind Cyclins-Cdks, nor inhibit cell growth or induce apoptosis. In the patient's tumor tissues, p27P69L associates with reduced/absent p27 expression and in one tumor with loss-of heterozygosity.Our results extend previous findings of CDKN1B mutations in patients with MEN1-related states and support the hypothesis of a tumor suppressor role for p27 in neuroendocrine cells.
Assuntos
Mutação em Linhagem Germinativa , Peptídeos e Proteínas de Sinalização Intracelular/genética , Neoplasia Endócrina Múltipla/genética , Idoso , Linhagem Celular , Inibidor de Quinase Dependente de Ciclina p27 , Análise Mutacional de DNA , DNA de Neoplasias/genética , Feminino , Técnicas de Silenciamento de Genes , Células HeLa , Humanos , Técnicas In Vitro , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Masculino , Neoplasia Endócrina Múltipla/classificação , Neoplasia Endócrina Múltipla Tipo 1/genética , Mutagênese Sítio-Dirigida , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Fenótipo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
CONTEXT: Approximately one-third of patients with acromegaly have concomitant hypertension. The outcome of hypertension after treatment of acromegaly is unknown. OBJECTIVE: To evaluate the role of GH and IGF-I control on systolic (SBP) and diastolic blood pressure (DBP) levels. PATIENTS: One hundred and five hypertensive patients (60 women, 45 men) with active disease receiving treatment for hypertension at their diagnosis of acromegaly. DESIGN: Observational, retrospective, multicentre. MEASUREMENTS: At diagnosis and after 24 months (median) of treatment we measured serum GH and IGF-I levels, blood pressure levels, left ventricular (LV) mass index (LVMi), early-to-late mitral flow velocity (E/A, as a measure of diastolic function) and LV ejection fraction (LVEF, as a measure of systolic function). RESULTS: At the diagnosis of acromegaly, hypertension was mild in 41.1% and severe in 58.9%. Serum GH and IGF-I levels did not differ in patients with mild or severe hypertension. After 24 months of treatment, all patients had a notable decrease in both GH and IGF-I levels, and achieved significantly lower levels of DBP, heart rate and LVMi; 76 patients (71%) had achieved control of GH and IGF-I levels. Only the patients with controlled acromegaly achieved significantly lower SBP levels and significantly improved cardiac systolic and diastolic function. A higher dose of antihypertensive drugs and/or an increased number of drugs to control hypertension were significantly greater in patients with uncontrolled (32.3%) than in those with controlled acromegaly (7.8%; P = 0.004). CONCLUSION: Hypertensive patients with controlled acromegaly achieved improved control of hypertension and of cardiac diastolic and systolic function. The use of antihypertensive drugs was significantly less in patients achieving control of acromegaly.
Assuntos
Acromegalia/fisiopatologia , Anti-Hipertensivos/uso terapêutico , Pressão Sanguínea/efeitos dos fármacos , Hormônio do Crescimento Humano/sangue , Hipertensão/tratamento farmacológico , Fator de Crescimento Insulin-Like I/metabolismo , Acromegalia/sangue , Acromegalia/complicações , Acromegalia/terapia , Adulto , Idoso , Idoso de 80 Anos ou mais , Inibidores da Enzima Conversora de Angiotensina/administração & dosagem , Inibidores da Enzima Conversora de Angiotensina/uso terapêutico , Anti-Hipertensivos/administração & dosagem , Diuréticos/administração & dosagem , Feminino , Hormônio do Crescimento Humano/metabolismo , Humanos , Hipertensão/complicações , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto JovemRESUMO
Breast cancer cells are usually sensitive to several chemotherapeutic regimens, but they can develop chemoresistance after prolonged exposure to cytotoxic drugs, acquiring a more aggressive phenotype. Drug resistance might involve the multi-drug resistance (MDR) 1 gene, encoding a transmembrane glycoprotein p-170 (P-gp), which antagonizes intracellular accumulation of cytotoxic agents, such as doxorubicin. We previously demonstrated that type 2 cyclooxygenase (COX-2) inhibitors can reverse the chemoresistance phenotype of a medullary thyroid carcinoma cell line by inhibiting P-gp expression and function. The aim of our study was to investigate the role of COX-2 inhibitors in modulating chemoresistance in a human breast cancer cell line, MCF7. MCF7 cells, expressing COX-2 but not MDR1, were treated with increasing doses of doxorubicin, and they became chemoresistant after 10 days of treatment, in association with MDR1 expression induction. This effect was reversed by doxorubicin withdrawal and prevented by co-incubation with N-[2-(cyclohexyloxy)4-nitrophenyl]-methanesulfonamide (NS-398), a selective COX-2 inhibitor. Treatment with NS-398 alone did not influence cell viability of a resistant MCF7 cell clone (rMCF7), but sensitized rMCF7 cells to the cytotoxic effects of doxorubicin. Moreover, treatment with NS-398 significantly reduced MDR1 expression in rMCF7 cells. Doxorubicin-induced membrane P-gp expression and function was also greatly impaired. Our data therefore support the hypothesis that COX-2 inhibitors can prevent or reduce the development of the chemoresistance phenotype in breast cancer cells by inhibiting P-gp expression and function.
Assuntos
Neoplasias da Mama/genética , Inibidores de Ciclo-Oxigenase 2/farmacologia , Ciclo-Oxigenase 2/genética , Resistencia a Medicamentos Antineoplásicos/fisiologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glicoproteínas/genética , Nitrobenzenos/farmacologia , Sulfonamidas/farmacologia , Subfamília B de Transportador de Cassetes de Ligação de ATP , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase/farmacologia , Doxorrubicina/farmacocinética , Doxorrubicina/toxicidade , Feminino , Humanos , Fenótipo , RNA Neoplásico/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Elevated circulating chromogranin A (CgA) levels are found in neuroendocrine tumors (NETs), but the diagnostic usefulness of this marker is still debatable. To assess the role of CgA for the diagnosis of gastroenteropancreatic (GEP) NETs and the identification of metastatic patients, an Italian multicenter observational study has been performed. CgA was evaluated in 202 GEP NET patients by IRMA and ELISA. The cutoffs for diagnosis and presence of metastases were identified by receiver-operating characteristic (ROC) curve. We found good correlation between IRMA and ELISA. The ROC analysis identified a cutoff of 53 ng/ml for IRMA and 16 U/l for ELISA as discriminating between controls and patients with active disease (sensitivity 71.3 and 84%; specificity 71 and 85% respectively). Metastases were present in 123 patients, having significantly higher CgA levels than patients without metastases. ROC analysis identified a cutoff of 146 ng/ml for IRMA and 67.3 U/l for ELISA as discriminating between patients with and without metastases (sensitivity 57 and 63.3%; specificity 55.6 and 71.4% respectively). For pancreatic NETs positive and negative predictive values were 84 and 78% respectively (90% specificity and 68% sensitivity). We found lower CgA levels in patients with extensive metastatic spread than in those with liver metastases only. These data assess the role of CgA evaluation in GEP NETs, and demonstrate that higher CgA levels associate with metastatic disease, confirming that CgA levels can provide a helpful practical biochemical marker for the clinical management of NETs, but with low sensitivity and specificity.
Assuntos
Biomarcadores Tumorais/sangue , Cromogranina A/sangue , Neoplasias Gastrointestinais/diagnóstico , Tumores Neuroendócrinos/diagnóstico , Neoplasias Pancreáticas/diagnóstico , Adulto , Ensaio de Imunoadsorção Enzimática , Feminino , Neoplasias Gastrointestinais/patologia , Humanos , Itália , Masculino , Pessoa de Meia-Idade , Tumores Neuroendócrinos/secundário , Neoplasias Pancreáticas/patologiaRESUMO
Somatostatin (SRIF) analogs have been employed in medical therapy of non-functioning pituitary adenomas (NFA), with contrasting results. Previous evidence showed that SRIF can exert its antiproliferative effects by reducing vascular endothelial growth factor (VEGF) secretion and action, and that VEGF expression may be related to pituitary tumor growth. The aim of our study was to clarify the possible effects of a multireceptor SRIF ligand on VEGF secretion and cell proliferation in human NFA primary cultures. We assessed the expression of SRIF receptors (SSTR1-5), the in vitro effects on VEGF secretion, and on cell viability of SRIF and of the stable SRIF analog pasireotide (SOM230), which activates SSTR1, 2, 3, and 5. Twenty-five NFA were examined by RT-PCR for expression of alpha-subunit, SSTR, VEGF, and VEGF receptors 1 (VEGF-R1) and 2 (VEGF-R2). Primary cultures were tested with SRIF and with pasireotide. All NFA samples expressed alpha-sub, VEGF and VEGFR-1 and 2, while SSTR expression pattern was highly variable. Two different groups were identified according to VEGF secretion inhibition by SRIF. VEGF secretion and cell viability were reduced by SRIF and pasireotide in the 'responder' group, but not in the 'non-responder' group, including NFA expressing SSTR5. SRIF and pasireotide completely blocked forskolin-induced VEGF secretion. In addition, SRIF and pasireotide completely abrogated the promoting effects of VEGF on NFA cell viability. Our data demonstrate that pasireotide can inhibit NFA cell viability by inhibiting VEGF secretion, and suggest that the multireceptor-SSTR agonist pasireotide might be useful in medical therapy of selected NFA.
Assuntos
Adenoma/metabolismo , Oligopeptídeos/farmacologia , Neoplasias Hipofisárias/metabolismo , Somatostatina/farmacologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto , Idoso , Sobrevivência Celular/efeitos dos fármacos , Feminino , Hormônios/farmacologia , Humanos , Ligantes , Masculino , RNA Mensageiro/metabolismo , Receptores de Somatostatina/genética , Receptores de Somatostatina/metabolismoRESUMO
Corticotropin-releasing hormone (CRH) is a key mediator of endocrine, autonomic, behavioral, and immune responses to stress. The ability of CRH to induce hormonal stress responses has been used to investigate the functionality of the hypothalamus-pituitary-adrenal axis, and consequently the activity of hypothalamic CRH neuronal systems. Indeed, CRH administration to humans causes prompt release of ACTH, followed by secretion of cortisol, aldosterone and other adrenal steroids. CRH hypersecretion/hyperactivity has been associated to major depression, anxiety-related disorders, anorexia nervosa, Alzheimer's and Parkinson's diseases and progressive supranuclear palsy. During pregnancy the human placenta and its accessory membranes are the major sites of CRH synthesis and secretion. Placental CRH secretion is autonomous, but increasing evidence indicates that maternal or fetal conditions may influence such secretion. Therefore, the emerging concept is that in the event of acute or chronic metabolic, physical or infectious stress, the placenta takes part in a stress syndrome by releasing CRH, which may contribute to restore local blood flow and to influence the timing of delivery. The CRH released by the placenta is measurable in maternal plasma and other biological fluids and may be used to diagnose subclinical processes anteceding pregnancy complications such as pre-eclampsia and preterm delivery.
Assuntos
Hormônio Liberador da Corticotropina/análise , Transtornos Mentais/diagnóstico , Trabalho de Parto Prematuro/diagnóstico , Pré-Eclâmpsia/diagnóstico , Hormônio Adrenocorticotrópico/sangue , Biomarcadores/análise , Biomarcadores/sangue , Hormônio Liberador da Corticotropina/sangue , Hormônio Liberador da Corticotropina/metabolismo , Feminino , Humanos , Doenças Neurodegenerativas/diagnóstico , GravidezRESUMO
Growth Hormone may influence neoplastic development of endometrial epithelium towards endometrial adenocarcinoma, which is one of the most occurring tumors in acromegalic patients. Since chemoresistance often develops in advanced endometrial adenocarcinoma, we investigated whether Growth Hormone might influence the development of chemoresistance to drugs routinely employed in endometrial adenocarcinoma treatment, such as Doxorubicin, Cisplatin, and Paclitaxel. Growth Hormone and Growth Hormone receptor expression was assessed by immunofluorescence in two endometrial adenocarcinoma cell lines, AN3 CA and HEC-1-A cells. Growth Hormone effects were assessed investigating cell viability, caspase3/7 activation, ERK1/2, and protein kinase C delta protein expression. AN3 CA and HEC-1-A cells display Growth Hormone and Growth Hormone receptor. Growth Hormone does not influence cell viability in both cells lines, but significantly reduces caspase 3/7 activation in AN3 CA cells, an effect blocked by a Growth Hormone receptor antagonist. Growth Hormone rescues AN3 CA cells from the inhibitory effects of Doxorubicin and Cisplatin on cell viability, while it has no effect on Paclitaxel. Growth Hormone does not influence the pro-apoptotic effects of Doxorubicin, but is capable of rescuing AN3 CA cells from the pro-apoptotic effects of Cisplatin. On the other hand, Growth Hormone did not influence the effects of Doxorubicin and Paclitaxel on HEC-1A cell viability. The protective action of Growth Hormone towards the effects of Doxorubicin may be mediated by ERK1/2 activation, while the pro-apoptotic effects of Cisplatin may be mediated by protein kinase C delta inhibition. All together our results indicate that Growth Hormone may differentially contribute to endometrial adenocarcinoma chemoresistance. This may provide new insights on novel therapies against endometrial adenocarcinoma chemoresistant aggressive tumors.
Assuntos
Adenocarcinoma/tratamento farmacológico , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cisplatino/farmacologia , Doxorrubicina/farmacologia , Neoplasias do Endométrio/tratamento farmacológico , Hormônio do Crescimento Humano/análogos & derivados , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Antineoplásicos/uso terapêutico , Antineoplásicos Hormonais/farmacologia , Antineoplásicos Hormonais/uso terapêutico , Caspase 3/metabolismo , Caspase 7/metabolismo , Linhagem Celular Tumoral , Cisplatino/uso terapêutico , Doxorrubicina/uso terapêutico , Resistencia a Medicamentos Antineoplásicos , Neoplasias do Endométrio/metabolismo , Neoplasias do Endométrio/patologia , Feminino , Hormônio do Crescimento Humano/metabolismo , Hormônio do Crescimento Humano/farmacologia , Humanos , Sinais de Exportação Nuclear/efeitos dos fármacos , Paclitaxel/farmacologia , Paclitaxel/uso terapêutico , Receptores da Somatotropina/metabolismoRESUMO
Somatostatin (SRIH) inhibits cell proliferation by interacting with five distinct SRIH receptor subtypes (SSTRs) activating several pathways in many tissues. We previously demonstrated that SRIH, by activating Src homology-2-containing protein, inhibits cell proliferation of the human medullary thyroid carcinoma cell line, TT, which expresses all SSTRs. However, the effects of SRIH on cell cycle proteins have not been investigated so far. We therefore evaluated the effects of SRIH and a selective SSTR2 agonist on cell cycle protein expression, mainly focusing on cyclin D1 and its associated kinases. Our data show that SRIH and the selective SSTR2 agonist, BIM-23120, reduce cell proliferation and DNA synthesis as well as induce a delay of the cell cycle in G(2)/M phase. Moreover, treatment with both SRIH and BIM-23120 decreases cyclin D1 levels, with a parallel increase in phosphocyclin D1 levels, suggesting protein degradation. Moreover, our data show an increase in glycogen synthase kinase-3beta activity, which triggers phosphorylation-dependent cyclin D1 degradation. Indeed, we observed a reduction in cyclin D1 protein half-life under treatment with SRIH or the SSTR2 selective agonist. A reduction in cdk4 protein levels is also observed with a parallel reduction in Rb phosphorylation levels at Ser-780. Our data indicate that the subtype 2 receptor-mediated antiproliferative effect of SRIH on TT cell proliferation may be exerted through a decrease in cyclin D1 levels.
Assuntos
Ciclina D1/metabolismo , Quinase 4 Dependente de Ciclina/metabolismo , Receptores de Somatostatina/metabolismo , Somatostatina/metabolismo , Neoplasias da Glândula Tireoide/metabolismo , Divisão Celular , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , DNA/metabolismo , Fase G2 , Quinase 3 da Glicogênio Sintase/metabolismo , Glicogênio Sintase Quinase 3 beta , Humanos , Técnicas In Vitro , Proteína do Retinoblastoma/metabolismoRESUMO
CONTEXT: The recognition of thyroid microcarcinoma has increased due to the widespread use of ultrasound-guided fine-needle aspiration biopsies. OBJECTIVE: The objective of this study was to describe histological and clinical characteristics of papillary thyroid microcarcinoma (PTMC) less than or equal to 1 cm. DESIGN: This study was a retrospective cohort. SETTING: This study was conducted at a university hospital endocrine clinic. PATIENTS: Over a 9-yr period, 243 consecutive patients with PTMC were studied. RESULTS: PTMC was an incidental finding at surgery in 21.4% of the PTMC cases. There were no differences in the clinical characteristics between those with incidental PTMC and those with suspected thyroid carcinoma. None of the patients with a cancer less than 8 mm had distant metastases, whereas distant metastases were observed in patients with cancers >/= 8 mm (P = 0.05). Disease-related mortality was not observed. CONCLUSIONS: PTMC is prevalent in the population. Among patients with PTMC, tumor size more than 8 mm is associated with more aggressive disease.
Assuntos
Carcinoma Papilar/patologia , Neoplasias da Glândula Tireoide/patologia , Biópsia por Agulha , Carcinoma Papilar/terapia , Humanos , Radioisótopos do Iodo/uso terapêutico , Metástase Linfática , Estudos Retrospectivos , Neoplasias da Glândula Tireoide/terapia , TireoidectomiaRESUMO
CONTEXT: The pathogenesis of increased blood pressure (BP) in acromegaly is unclear, and the role of IGF-I levels and the renin-angiotensin-aldosterone system (RAAS) in this disease remains controversial. OBJECTIVE AND DESIGN: The aim of this study was to investigate the role of gene polymorphisms of the RAAS and involved in sodium handling on BP in acromegaly. SETTING AND PATIENTS: We conducted a multicentric retrospective study that included 100 consecutive patients with acromegaly referred during the period 2000-2003. INTERVENTION: All patients were genotyped for ACE I/D, AGT M235T, CYP11B2 -344T/C, B2R -58T/C, and alpha-adducin G460W polymorphisms. MAIN OUTCOME MEASURE: We assessed the prevalence of hypertension and BP according to the genotype. RESULTS: Patients with the CYP11B2 -344CC genotype displayed a significant increase in the risk of hypertension compared with patients with CT/TT genotypes (odds ratio = 4.0; 95% confidence interval = 1.4-11.6; P = 0.01). Consistently, a significant proportion of patients with the CYP11B2 -344CC genotypes were under antihypertensive treatment (73.1%) compared with patients with the TT/TC genotypes (38.2%; P = 0.003). Patients with the -344CC genotype displayed a significant increase in systolic BP (10.2 +/- 4.3 mm Hg; P = 0.02) but not a significant increase in diastolic BP (2.6 +/- 2.6 mm Hg; P = 0.32) compared with patients with the CT/TT genotype. CONCLUSIONS: We have shown an association of the -344T/C CYP11B2 gene polymorphism with BP in patients affected by acromegaly. These findings suggest that the RAAS is implicated in the pathogenesis of hypertension in acromegaly.
Assuntos
Acromegalia/genética , Acromegalia/fisiopatologia , Pressão Sanguínea/fisiologia , Citocromo P-450 CYP11B2/genética , Polimorfismo Genético , Adulto , Idoso , Estudos Transversais , Feminino , Frequência do Gene , Genótipo , Humanos , Hipertensão/epidemiologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de RiscoRESUMO
CONTEXT: Cerebrovascular disease is highly prevalent in the general population, frequently leading to permanent invalidity and reduced quality of life. IGF-I is recognized as an important neuroprotective factor against cerebral hypoxic insult. OBJECTIVE: The objective of the study was to evaluate pituitary function, in particular GH-IGF-I axis, in adult patients receiving rehabilitation after an ischemic stroke. SUBJECTS AND METHODS: We studied 42 patients (12 females; age range, 50-88 yr) during rehabilitation after stroke, evaluating the relationship between the GH-IGF-I axis and the severity (National Institutes of Health stroke scale) and outcome [Rancho Los Amigos Scale of Cognitive Functioning (LCFS); Functional Independence Measure (FIM); modified Ranking Scale] from stroke. RESULTS: GH deficiency was demonstrated in five patients (11.9%). Peak GH after GHRH + arginine test and IGF-I levels did not correlate with severity of stroke. IGF-I was positively correlated with LCFS (r = 0.305, P < 0.05) and the difference between FIM on admission and at discharge from rehabilitation (DeltaFIM; r = 0.361, P < 0.02). Outcome indexes (LCFS, FIM at discharge, DeltaFIM) and occurrence of favorable outcome (modified Ranking Scale 0-1) were significantly (P < 0.05) higher in patients with IGF-I levels 161.8 mug/dl or greater (50th percentile of the patient distribution). LH-FSH deficiency (three cases), ACTH deficiency (one case), and hyperprolactinemia (two cases) were detected. One patient had primary hypogonadism, and six males had low testosterone with normal LH and FSH levels. By multivariate analysis, IGF-I level was the main significant predictor of DeltaFIM and LCFS. CONCLUSIONS: Ischemic stroke may be associated with pituitary dysfunction, particularly GH and gonadotropin deficiencies. The higher IGF-I levels observed in patients with better outcome suggest a possible neuroprotective role of IGF-I. Circulating IGF-I may predict functional performance during rehabilitation and ischemic stroke outcome.
Assuntos
Isquemia Encefálica/sangue , Fator de Crescimento Insulin-Like I/análise , Acidente Vascular Cerebral/sangue , Hormônio Adrenocorticotrópico/sangue , Idoso , Idoso de 80 Anos ou mais , Feminino , Hormônio do Crescimento Humano/deficiência , Humanos , Masculino , Pessoa de Meia-Idade , Hipófise/fisiopatologia , Valor Preditivo dos TestesRESUMO
CONTEXT: Medullary thyroid carcinoma (MTC) is a rare tumor originating from thyroid parafollicular C cells. We previously demonstrated that somatostatin (SRIH) reduces cell growth in the human MTC cell line, TT, which expresses all SRIH receptor (SSTR) subtypes and responds differently to selective SSTR agonists. OBJECTIVE: To clarify the possible effects of SRIH analogs on hormone secretion and proliferation in MTC primary cultures, we evaluated SSTR expression and assessed the in vitro effects on calcitonin (CT) and chromogranin A secretion as well as cell viability of SRIH analogs interacting with SSTR1, SSTR2, and SSTR5. DESIGN: Thirty-five patients affected by MTC were recruited from 2003 to 2005. After total thyroidectomy, the samples were examined for CT, chromogranin A, and SSTR expression by RT-PCR. Primary cultures were developed and tested with SRIH analogs interacting with SSTR1, SSTR2, and SSTR5. RESULTS: We selected 18 MTC tumor samples, expressing SSTR1, SSTR2, and SSTR5. Two different groups were identified according to CT secretion inhibition by the clinically available SRIH analog, lanreotide. In the responder group, CT secretion was reduced by compounds interacting with SSTR1, SSTR2, and SSTR5, whereas cell viability was not affected. On the other hand, in the nonresponder group, CT secretion was reduced by the SSTR1 selective agonist, whereas cell viability was inhibited by SSTR2 selective agonists. CONCLUSIONS: Our data suggest that SRIH analogs might be useful in medical therapy of MTC because they could have antiproliferative effects despite the lack of antisecretory activity and vice versa.
Assuntos
Calcitonina/metabolismo , Carcinoma Medular/tratamento farmacológico , Cromograninas/metabolismo , Receptores de Somatostatina/agonistas , Neoplasias da Glândula Tireoide/tratamento farmacológico , Adulto , Idoso , Carcinoma Medular/metabolismo , Carcinoma Medular/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Criança , Cromogranina A , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Receptores de Somatostatina/análise , Receptores de Somatostatina/classificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias da Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/patologiaRESUMO
Protein Kinase C Delta (PRKCD) has been highlighted among disrupted pathways in corticotroph adenomas. PRKCD is expressed at low level in human corticotroph adenomas and controls cell cycle in vitro. Therefore, PRKCD may play an important role in the development/progression of corticotroph adenomas, warranting further studies to understand the role of PRKCD and related pathways in restraining pituitary cell growth. We evaluated PRKCD role in influencing cell behavior in terms of cell viability, hormone expression and protein expression profile, by silencing PRKCD in AtT-20/D16v-F2 cells. PRKCD silencing increases cell viability, enhances hormone expression and induces morphological changes associated with deregulation of adhesion molecules. PRKCD silencing is associated with an increase in Epithelial Growth Factor Receptor (EGFR) expression, a marker of tumor aggressive behavior, and sensitivity to anti-EGFR molecules. PRKCD might restrain corticotroph adenoma cells from acquiring an aggressive behavior, candidating PRKCD as a possible molecular target for the treatment of corticotroph adenomas.
Assuntos
Hormônio Adrenocorticotrópico/metabolismo , Neoplasias Hipofisárias/metabolismo , Proteína Quinase C-delta/metabolismo , Animais , Linhagem Celular Tumoral , Sobrevivência Celular , Receptores ErbB/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Camundongos , Pró-Opiomelanocortina/metabolismoRESUMO
Medullary thyroid carcinoma (MTC) is a rare tumor originating from thyroid parafollicular C cells, where, in the inherited form, constitutive activation of the RET protooncogene is responsible for unrestrained cell proliferation. We previously demonstrated that somatostatin (SRIF) reduces cell growth in the human MTC cell line TT, which expresses all SRIF receptor (SSTR) subtypes and responds differently to selective SSTR agonists. The antiproliferative mechanism of SRIF and its analogs in MTC is still unclear. Src homology-2-containing protein tyrosine phosphatase-1 (SHP-1), a cytoplasmic protein tyrosine phosphatase (PTP), is activated by somatotropin release-inhibiting factor and reduces mutated RET autophosphorylation in a heterologous system. In this study, we explore the role of PTP activation, in particular of SHP-1, in TT cells, where RET is constitutively activated. In TT cells, SRIF stimulated the PTP activity of SHP-1, which was associated with proliferation inhibition and with reduction in the MAPK pathway activation. Blockade of PTP activity with sodium orthovanadate induced cell proliferation and MAPK phosphorylation and blunted the inhibitory effects of SRIF. Moreover, SHP-1 associates with SSTR2 depending on its activation. By using a MAPK kinase inhibitor, we demonstrated that TT cell growth depends on MAPK pathway activation. Furthermore, in TT cells overexpressing SHP-1, cell proliferation and MAPK signaling were strongly down-regulated, whereas in TT cells transfected with a dominant negative form of SHP-1, cell proliferation and MAPK signaling were markedly induced. Our data demonstrate that SRIF inhibitory effects on TT cell proliferation are mediated, at least in part, by SHP-1, which acts through a MAPK-dependent mechanism.
Assuntos
Carcinoma Medular/metabolismo , Carcinoma Medular/patologia , Proteínas Tirosina Fosfatases/metabolismo , Neoplasias da Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/patologia , Divisão Celular/fisiologia , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Sistema de Sinalização das MAP Quinases/fisiologia , Fosforilação , Proteína Fosfatase 1 , Proteína Tirosina Fosfatase não Receptora Tipo 6 , Proteínas Tirosina Fosfatases/genética , Receptores de Somatostatina/metabolismoRESUMO
A 29-yr-old woman presented with acromegaly, pituitary gland enlargement, and an isolated pulmonary mass of 3.3 cm in diameter, which displayed a very high tracer uptake after OctreoScan. Plasma GHRH levels were markedly elevated. The patient underwent left lung upper lobectomy, and histopathology disclosed a bronchial atypical carcinoid. The tissue was examined for somatostatin (SRIH) receptor subtypes (SSTRs) 1-5 expression by RT-PCR. Cultured tumor cells were treated with SRIH, lanreotide (BIM-23014), or SRIH analogs selective for SSTR2 (BIM-23120), SSTR5 (BIM-23206), or SSTR1 (BIM-23926). GHRH was measured in the medium after 6 h, and cell viability was assessed after 48 h. RT-PCR analysis showed expression of SSTR1, -2, and -5. GHRH secretion was significantly reduced by SRIH (-50%), Lan (-35%), as well as by the SSTR2, SSTR5, and SSTR1 selective agonists (-55, -75, and -20%, respectively), whereas cell viability was not affected. Our data show SSTR expression in a GHRH-secreting bronchial carcinoid and provide evidence that, in vitro, selective SSTR activation differently inhibit ectopic GHRH secretion. These findings suggest that SSTR-specific SRIH analogs may be useful in the medical therapy of GHRH-secreting bronchial carcinoids.
Assuntos
Neoplasias Brônquicas/tratamento farmacológico , Tumor Carcinoide/tratamento farmacológico , Hormônio Liberador de Hormônio do Crescimento/metabolismo , Receptores de Somatostatina/agonistas , Adulto , Neoplasias Brônquicas/metabolismo , Neoplasias Brônquicas/patologia , Tumor Carcinoide/metabolismo , Tumor Carcinoide/patologia , Sobrevivência Celular/efeitos dos fármacos , Feminino , Humanos , Receptores de Somatostatina/classificaçãoRESUMO
OBJECTIVE: Medullary thyroid carcinoma (MTC) is a highly chemoresistant malignant neoplasia deriving from parafollicular C cells. Chemotherapy failure has been ascribed, at least in part, to the overexpression by MTC of the multidrug resistance 1 (MDR1) gene, encoding a transmembrane glycoprotein [permeability glycoprotein (P-gp)] that antagonizes intracellular accumulation of cytotoxic agents. P-gp expression and function in a rat model have been demonstrated to depend on cyclooxygenase (COX)-2 isoform levels, which are found elevated in many human cancers. The aim of our study was to investigate the role of the COX-2 pathway in modulating chemoresistance. DESIGN AND RESULTS: We investigated P-gp and COX-2 expression and then evaluated the sensitizing effects of COX-2 inhibitors on the cytotoxic effects of doxorubicin in the presence or in the absence of prostaglandin E2 in primary cultures and in a human MTC cell line, TT. Moreover, P-gp function has been studied. Our data show that TT cells express both MDR1 and COX-2 and that rofecoxib, a selective COX-2 inhibitor, sensitizes TT cells to the cytotoxic effects of doxorubicin, reducing P-gp expression and function. CONCLUSIONS: Our data suggest that these effects are mediated by a mechanism not involving the generation of prostaglandin E2, possibly implicating the synthesis of other COX-2 products.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/fisiologia , Carcinoma Medular/tratamento farmacológico , Carcinoma Medular/metabolismo , Inibidores de Ciclo-Oxigenase/farmacologia , Resistencia a Medicamentos Antineoplásicos , Prostaglandina-Endoperóxido Sintases , Neoplasias da Glândula Tireoide/tratamento farmacológico , Neoplasias da Glândula Tireoide/metabolismo , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/biossíntese , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/efeitos dos fármacos , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Antineoplásicos/farmacologia , Western Blotting , Bloqueadores dos Canais de Cálcio/farmacologia , Contagem de Células , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ciclo-Oxigenase 2 , Inibidores de Ciclo-Oxigenase 2 , Dinoprostona/farmacologia , Doxorrubicina/farmacologia , Humanos , Lactonas/farmacologia , Proteínas de Membrana , Permeabilidade , Fenótipo , RNA Neoplásico/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sulfonas/farmacologia , Verapamil/farmacologiaRESUMO
Traumatic brain injury (TBI) is one of the main causes of death and disability in young adults, with consequences ranging from physical disabilities to long-term cognitive, behavioural, psychological and social defects. Post-traumatic hypopituitarism (PTHP) was recognized more than 80 years ago, but it was thought to be a rare occurrence. Recently, clinical evidence has demonstrated that TBI may frequently cause hypothalamic-pituitary dysfunction, probably contributing to a delayed or hampered recovery from TBI. Changes in pituitary hormone secretion may be observed during the acute phase post-TBI, representing part of the acute adaptive response to the injury. Moreover, diminished pituitary hormone secretion, caused by damage to the pituitary and/or hypothalamus, may occur at any time after TBI. PTHP is observed in about 40% of patients with a history of TBI, presenting as an isolated deficiency in most cases, and more rarely as complete pituitary failure. The most common alterations appear to be gonadotropin and somatotropin deficiency, followed by corticotropin and thyrotropin deficiency. Hyper- or hypoprolactinemia may also be present. Diabetes insipidus may be frequent in the early, acute phase post-TBI, but it is rarely permanent. Severity of TBI seems to be an important risk factor for developing PTHP; however, PTHP can also manifest after mild TBI. Accurate evaluation and long-term follow-up of all TBI patients are necessary in order to detect the occurrence of PTHP, regardless of clinical evidence for pituitary dysfunction. In order to improve outcome and quality of life of TBI patients, an adequate replacement therapy is of paramount importance.
Assuntos
Lesões Encefálicas/complicações , Lesões Encefálicas/fisiopatologia , Hipopituitarismo/etiologia , Hipopituitarismo/fisiopatologia , HumanosRESUMO
Chronic growth hormone (GH)/insulin-like growth factor I (IGF-I) excess is associated with considerable mortality in acromegaly, but no data are available in pituitary gigantism. The aim of the study was to evaluate the long-term effects of early exposure to GH and IGF-I excess on cardiovascular and metabolic parameters in adult patients with pituitary gigantism. Six adult male patients with newly diagnosed gigantism due to GH secreting pituitary adenoma were studied and compared with 6 age- and sex-matched patients with acromegaly and 10 healthy subjects. Morphologic and functional cardiac parameters were evaluated by Doppler echocardiography. Glucose metabolism was assessed by evaluating glucose tolerance and homeostasis model assessment index. Disease duration was significantly longer (P<.05) in patients with gigantism than in patients with acromegaly, whereas GH and IGF-I concentrations were comparable. Left ventricular mass was increased both in patients with gigantism and in patients with acromegaly, as compared with controls. Left ventricular hypertrophy was detected in 2 of 6 of both patients with gigantism and patients with acromegaly, and isolated intraventricular septum thickening in 1 patient with gigantism. Inadequate diastolic filling (ratio between early and late transmitral flow velocity<1) was detected in 2 of 6 patients with gigantism and 1 of 6 patients with acromegaly. Impaired glucose metabolism occurrence was higher in patients with acromegaly (66%) compared with patients with gigantism (16%). Concentrations of IGF-I were significantly (P<.05) higher in patients with gigantism who have cardiac abnormalities than in those without cardiac abnormalities. In conclusion, our data suggest that GH/IGF-I excess in young adult patients is associated with morphologic and functional cardiac abnormalities that are similar in patients with gigantism and in patients with acromegaly, whereas occurrence of impaired glucose metabolism appears to be higher in patients with acromegaly, although patients with gigantism are exposed to GH excess for a longer period.
Assuntos
Gigantismo/complicações , Gigantismo/metabolismo , Hormônio do Crescimento Humano/sangue , Hipertrofia Ventricular Esquerda/etiologia , Hipertrofia Ventricular Esquerda/metabolismo , Fator de Crescimento Insulin-Like I/metabolismo , Acromegalia/complicações , Acromegalia/metabolismo , Adulto , Pressão Sanguínea , Ecocardiografia Doppler , Eletrocardiografia , Glucose/metabolismo , Intolerância à Glucose/etiologia , Intolerância à Glucose/metabolismo , Humanos , Hipertrofia Ventricular Esquerda/diagnóstico por imagem , Masculino , Disfunção Ventricular Esquerda/diagnóstico por imagem , Disfunção Ventricular Esquerda/etiologia , Disfunção Ventricular Esquerda/metabolismoRESUMO
In the present study we examined the clinical value of a differential response of thyroglobulin (Tg) concentration after recombinant human thyrotropin (rhTSH) stimulation (rhTSH Tg testing) and its correlation with (131)I uptake and whole-body scanning (rhTSH-WBS) in 104 patients who had previously undergone near-total thyroidectomy and (131)I ablation for differentiated thyroid carcinoma (DTC). rhTSH Tg testing was considered negative for rhTSH-Tg less than 0.9 ng/mL, low positive for rhTSH-Tg of 1-5 ng/mL and high positive for rhTSHTg greater than 5 ng/mL. rhTSH Tg testing was negative in 70 patients, 1 of whom had a lymph-node metastasis, but no (131)I uptake. Seven patients had low positive rhTSH Tg testing and no (131)I uptake, but 2 of these patients had cervical lymph node metastases. Twenty-seven patients had high positive rhTSH Tg testing and (131)I uptake was detected in lung, bone, or mediastinum in 11. Imaging techniques (computed tomography [CT], magnetic resonance imaging [MRI], fluorine-18 2-fluoro-2-deoxy-D-glucose-positron emission tomography [FDGPET]) documented metastatic disease in 22. In conclusion, our results suggest that any rise in rhTSH-Tg, even at low level, should raise the suspicion of persistent or recurrent DTC. Patients with rhTSH-Tg at high level should be carefully evaluated, because DTC persistence is highly probable. TSH-WBS provides little adjunctive information.