RESUMO
Although mitochondrial and serotonergic dysfunctions have been implicated in the etiology of bipolar disorder (BD), the relationship between these unrelated pathways has not been elucidated. A family of BD and chronic progressive external ophthalmoplegia (CPEO) caused by a mutation of the mitochondrial adenine nucleotide translocator 1 (ANT1, SLC25A4) implicated that ANT1 mutations confer a risk of BD. Here, we sequenced ANT1 in 324 probands of NIMH bipolar disorder pedigrees and identified two BD patients carrying heterozygous loss-of-function mutations. Behavioral analysis of brain specific Ant1 heterozygous conditional knockout (cKO) mice using lntelliCage showed a selective diminution in delay discounting. Delay discounting is the choice of smaller but immediate reward than larger but delayed reward and an index of impulsivity. Diminution of delay discounting suggests an increase in serotonergic activity. This finding was replicated by a 5-choice serial reaction time test. An anatomical screen showed accumulation of COX (cytochrome c oxidase) negative cells in dorsal raphe. Dorsal raphe neurons in the heterozygous cKO showed hyperexcitability, along with enhanced serotonin turnover in the nucleus accumbens and upregulation of Maob in dorsal raphe. These findings altogether suggest that mitochondrial dysfunction as the genetic risk of BD may cause vulnerability to BD by altering serotonergic neurotransmission.
Assuntos
Translocador 1 do Nucleotídeo Adenina/genética , Translocador 1 do Nucleotídeo Adenina/metabolismo , Transtorno Bipolar/genética , Animais , Transtorno Bipolar/metabolismo , Desvalorização pelo Atraso/fisiologia , Núcleo Dorsal da Rafe/metabolismo , Feminino , Humanos , Comportamento Impulsivo , Masculino , Camundongos , Camundongos Knockout , Mitocôndrias/genética , Mitocôndrias/metabolismo , Oftalmoplegia Externa Progressiva Crônica/metabolismo , Recompensa , Neurônios Serotoninérgicos/metabolismo , Neurônios Serotoninérgicos/fisiologiaRESUMO
Transgenic mouse models of Alzheimer's disease (AD) with nonphysiologic overexpression of amyloid precursor protein (APP) exhibit various unnatural symptoms/dysfunctions. To overcome this issue, mice with single humanized App knock-in (KI) carrying Swedish (NL), Beyreuther/Iberian (F), and Arctic (G) mutations in different combinations were recently developed. The validity of these mouse models of AD from a behavioral viewpoint, however, has not been extensively evaluated. Thus, using an automated behavior monitoring system, we analyzed various behavioral domains, including executive function, and learning and memory. The App-KI mice carrying NL-G-F mutations showed clear deficits in spatial memory and flexible learning, enhanced compulsive behavior, and reduced attention performance. Mice carrying NL-F mutations exhibited modest abnormalities. The NL-G-F mice had a greater and more rapid accumulation of Aß deposits and glial responses. These findings reveal that single pathologic App-KI is sufficient to produce deficits in broad cognitive domains and that App-KI mouse lines with different levels of pathophysiology are useful models of AD.
Assuntos
Doença de Alzheimer/fisiopatologia , Precursor de Proteína beta-Amiloide/genética , Comportamento Animal/fisiologia , Disfunção Cognitiva/fisiopatologia , Função Executiva/fisiologia , Aprendizagem/fisiologia , Doença de Alzheimer/genética , Animais , Modelos Animais de Doenças , Feminino , Masculino , Camundongos , Camundongos Transgênicos , Memória Espacial/fisiologiaRESUMO
Super-channel transmission is a promising solution to increase the capacity of a channel beyond 100 Gb/s in next-generation optical networks. The performance of a super-channel comprising multiple subcarriers, however, degrades if optical filtering distortions occur in particular subcarriers. In this paper, we propose a method that improves super-channel performance by dispersing the distortions over all subcarriers. We also numerically demonstrate that the method effectively mitigates the filtering-induced penalty suffered by super-channels.
RESUMO
Alzheimer's disease (AD) is a worldwide burden. Diagnosis is complicated by the fact that AD is asymptomatic at an early stage. Studies using AD-modeled animals offer important and useful insights. Here, we classified mice with a high risk of AD at a preclinical stage by using only their behaviors. Wild-type and knock-in AD-modeled (App NL-G-F/NL-G-F ) mice were raised, and their cognitive behaviors were assessed in an automated monitoring system. The classification utilized a machine learning method, i.e., a deep neural network, together with optimized stepwise feature selection and cross-validation. The AD risk could be identified on the basis of compulsive and learning behaviors (89.3% ± 9.8% accuracy) shown by AD-modeled mice in the early age (i.e., 8-12 months old) when the AD symptomatic cognitions were relatively underdeveloped. This finding reveals the advantage of machine learning in unveiling the importance of compulsive and learning behaviors for early AD diagnosis in mice.
RESUMO
In the current study, we established a novel murine ischemic brain damage model using a photochemical reaction to evaluate the recovery of neurological dysfunction and brain repair reactions. In this model, reproducible damage was induced in the frontal lobe of the cortex, which was accompanied by neurological dysfunction. Sequential changes in damage size, microglial accumulation, astrocyte activation, and neurological dysfunction were studied in C57BL/6J and BALB/c mouse strains. Although the initial size of damage was comparable in both strains, the extent of damage was later reduced to a greater extent in C57BL/6J mice than that in BALB/c mice. In addition, C57BL/6J mice showed later edema clearance until day 7, less microglial accumulation, and relatively more astrocyte activation on day 7. Neurologic dysfunction was evaluated by three behavioral tests: the von Frey test, the balance beam test, and the tail suspension test. The behavioral abnormalities evaluated by these tests were remarkable following the induction of damage and recovered by day 21 in both strains. However, the abnormalities were more prominent and the recovery was later in C57BL/6J mice. These findings demonstrate that our novel ischemic stroke model is useful for evaluating brain repair reactions and the recovery of neurological dysfunction in mice with different genetic backgrounds. In addition, we found that both the brain repair reactions and the recovery of neurological dysfunction after comparable ischemic brain damage varied between strains; in that, they both occurred later in C57BL/6J mice.
Assuntos
Isquemia Encefálica/fisiopatologia , AVC Isquêmico/fisiopatologia , Animais , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Especificidade da EspécieRESUMO
The hippocampus, a region critical for memory and spatial navigation, has been implicated in delay discounting, the decline in subjective reward value when a delay is imposed. However, how delay information is encoded in the hippocampus is poorly understood. Here, we recorded from CA1 of mice performing a delay-discounting decision-making task, where delay lengths, delay positions, and reward amounts were changed across sessions, and identified subpopulations of CA1 neurons that increased or decreased their firing rate during long delays. The activity of both delay-active and -suppressed cells reflected delay length, delay position, and reward amount; but manipulating reward amount differentially impacted the two populations, suggesting distinct roles in the valuation process. Further, genetic deletion of the N-methyl-D-aspartate (NMDA) receptor in hippocampal pyramidal cells impaired delay-discount behavior and diminished delay-dependent activity in CA1. Our results suggest that distinct subclasses of hippocampal neurons concertedly support delay-discounting decisions in a manner that is dependent on NMDA receptor function.
Assuntos
Comportamento Animal , Região CA1 Hipocampal/fisiologia , Desvalorização pelo Atraso , Animais , Região CA1 Hipocampal/química , Aprendizagem em Labirinto , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neurônios/fisiologia , Análise e Desempenho de TarefasRESUMO
Multiple factors-such as aging and genes-are frequently associated with cognitive decline. Genetically modified mouse models of cognitive decline, such as Alzheimer's disease (AD), have become a promising tool to elucidate the underlying mechanisms and promote the therapeutic advances. An important step is the validation and characterization of expected behavioral abnormality in the models, in the case of AD, cognitive decline. The long-term behavioral investigations of laboratory animals to study the effect of aging demand substantial efforts from researchers. The IntelliCage system is a high-throughput and cost-effective test battery for mice that eliminates the need for daily human handling. Here, we describe how the system is utilized in the long-term phenotyping of a genetic Alzheimer's disease model, specifically focusing on the cognitive functions. The experiment employs repeated battery of tests that assess spatial learning and executive functions. This cost-effective age-dependent phenotyping allows us to identify the transient and/or permanent effects of genes on various cognitive aspects.
Assuntos
Doença de Alzheimer/genética , Cognição/ética , Modelos Genéticos , Animais , Modelos Animais de Doenças , Humanos , Masculino , Camundongos , Camundongos TransgênicosRESUMO
Tail pinch facilitates eating in rats. We investigated an unidentified link between tail-pinch-induced eating behavior and individual emotionality in male Sprague-Dawley rats. Anxiety-like behavior was assessed on the elevated plus maze (EPM) and in the open field test (OFT). Tail-pinch-induced eating was observed as follows: After a 30-min habituation period, the tail pinch was applied for 5 min, followed by a 30-min recovery period. During the habituation and recovery periods, rats were allowed to access food ad libitum. During the recovery period, 14 of 24 rats ate more food than during the habituation period. Thus, we named them "high responders" and the others as "low responders". The food intake was significantly greater, while the times spent in the open arms in the EPM and in the center area in the OFT were significantly shorter in high responders than in low responders. This result suggests that the rats consuming more food after mild stress have higher anxiety.
Assuntos
Adaptação Psicológica/fisiologia , Ansiedade/fisiopatologia , Ingestão de Alimentos/psicologia , Emoções/fisiologia , Comportamento Alimentar/psicologia , Estresse Psicológico/fisiopatologia , Animais , Comportamento Animal/fisiologia , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Cadmium is an environmental electrophile that modifies reactive thiols in proteins, indicating that this heavy metal may modulate redox-signal transduction pathways. The current consensus is that reactive persulfides and polysulfides produced by cystathionine γ-lyase (CSE) and cystathionine ß-synthase are highly nucleophilic and thus cadmium may be captured by these reactive sulfur species. It has previously been found that electrophile-mediated covalent modifications of the heat shock protein (HSP) are involved in the activation of heat shock factor 1 (HSF1) pathway. The effects of cadmium on the activation of HSP/HSF1 pathway were investigated in this study. Exposure of bovine aortic endothelial cells to cadmium resulted in modification of HSP90 and HSF1 activation, thereby up-regulating the downstream protein HSP70. The siRNA-mediated knockdown of HSF1 enhanced the cytotoxicity induced by cadmium, suggesting that the HSP90/HSF1 pathway contributes to protection against cadmium toxicity. The knockdown of CSE and/or cystathionine ß-synthase decreased the levels of reactive sulfur species in the cells and increased the degree of HSP70 induction and cytotoxicity caused by exposure to cadmium. Overexpression of CSE diminished cadmium-mediated up-regulation of HSP70 and cytotoxicity. These results suggest that cadmium activates HSF1 by modifying HSP90 and that reactive sulfur species regulate the redox signal transduction pathway presumably via capture of cadmium, resulting in protection against cadmium toxicity under toxic conditions.
Assuntos
Cádmio/toxicidade , Células Endoteliais/efeitos dos fármacos , Proteínas de Choque Térmico HSP90/metabolismo , Fatores de Transcrição de Choque Térmico/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sulfetos/metabolismo , Animais , Bovinos , Técnicas de Cultura de Células , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Cistationina beta-Sintase/metabolismo , Cistationina gama-Liase/metabolismo , Células Endoteliais/metabolismoRESUMO
In our previous rodent studies, the paclitaxel (PTX)-incorporating polymeric micellar nanoparticle formulation NK105 had showed significantly stronger antitumor effects and reduced peripheral neurotoxicity than PTX dissolved in Cremophor® EL and ethanol (PTX/CRE). Thus, to elucidate the mechanisms underlying reduced peripheral neurotoxicity due to NK105, we performed pharmacokinetic analyses of NK105 and PTX/CRE in rats. Among neural tissues, the highest PTX concentrations were found in the dorsal root ganglion (DRG). Moreover, exposure of DRG to PTX (Cmax_PTX and AUC0-inf._PTX) in the NK105 group was almost half that in the PTX/CRE group, whereas exposure of sciatic and sural nerves was greater in the NK105 group than in the PTX/CRE group. In histopathological analyses, damage to DRG and both peripheral nerves was less in the NK105 group than in the PTX/CRE group. The consistency of these pharmacokinetic and histopathological data suggests that high levels of PTX in the DRG play an important role in the induction of peripheral neurotoxicity, and reduced distribution of PTX to the DRG of NK105-treated rats limits the ensuing peripheral neurotoxicity. In further analyses of PTX distribution to the DRG, Evans blue (Eb) was injected with BODIPY®-labeled NK105 into rats, and Eb fluorescence was observed only in the DRG. Following injection, most Eb dye bound to albumin particles of ~8 nm and had penetrated the DRG. In contrast, BODIPY®-NK105 particles of ~90 nm were not found in the DRG, suggesting differential penetration based on particle size. Because PTX also circulates as PTX-albumin particles of ~8 nm following injection of PTX/CRE, reduced peripheral neurotoxicity of NK105 may reflect exclusion from the DRG due to particle size, leading to reduced PTX levels in rat DRG (275).
Assuntos
Micelas , Nanopartículas/química , Neurotoxinas/toxicidade , Paclitaxel/análogos & derivados , Paclitaxel/farmacologia , Polímeros/química , Albuminas/metabolismo , Animais , Biomarcadores/metabolismo , Química Farmacêutica , Etanol/química , Azul Evans/metabolismo , Feminino , Gânglios Espinais/efeitos dos fármacos , Gânglios Espinais/patologia , Glicerol/análogos & derivados , Glicerol/química , Imuno-Histoquímica , Injeções , Paclitaxel/administração & dosagem , Paclitaxel/farmacocinética , Paclitaxel/toxicidade , Ratos Sprague-Dawley , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/patologiaRESUMO
Accumulating evidence suggests that a coordinately controlled G(2) checkpoint prevents cells with damaged DNA from entering mitosis, thus playing an important role in the maintenance of chromosomal integrity. In the study presented here, we identified a homozygous deletion of the 14-3-3epsilon gene, which resides within a previously identified, commonly deleted region at 17p13.3 in lung cancers, in two small cell lung cancer cell lines that originate from distinct metastatic sites of the same patients. The introduction of 14-3-3epsilon induced significantly restored G(2) checkpoint responses, which resulted in the reduction of mitotic cells as well as of aberrant mitotic figures in the X-ray-irradiated 14-3-3epsilon-null small cell lung cancer cell line. Interestingly, we also found that the G(2) checkpoint response is frequently impaired to various degrees in a large fraction of small cell lung cancer cell lines. These findings suggest the possible involvement of the perturbed G(2) checkpoint in the pathogenesis of this aggressive form of human lung cancers.
Assuntos
Carcinoma de Células Pequenas/genética , Cromossomos Humanos Par 17/genética , Fase G2/genética , Neoplasias Pulmonares/genética , Tirosina 3-Mono-Oxigenase/genética , Proteínas 14-3-3 , Carcinoma de Células Pequenas/patologia , Divisão Celular/genética , Deleção de Genes , Humanos , Neoplasias Pulmonares/patologia , Mitose/genética , Transfecção , Células Tumorais CultivadasRESUMO
It has been suggested that attenuation of the decatenation G(2) checkpoint function, which ensures sufficient chromatid decatenation by topoisomerase II before entering into mitosis, may contribute to the acquisition of genetic instability in cancer cells. To date, however, very little information is available on this type of checkpoint defect in human cancers. In this study, we report for the first time that a proportion of human lung cancer cell lines did not properly arrest before entering mitosis in the presence of a catalytic, circular cramp-forming topoisomerase II inhibitor ICRF-193, whereas the decatenation G(2) checkpoint impairment was present independently of the impaired DNA damage G(2) checkpoint. In addition, the presence of decatenation G(2) checkpoint dysfunction was found to be associated with diminished activation of ataxia-telangiectasia mutated in response to ICRF-193, suggesting the potential involvement of an upstream pathway sensing incompletely catenated chromatids. Interestingly, hypersensitivity to ICRF-193 was observed in cell lines with decatenation G(2) checkpoint impairment and negligible activation of ataxia-telangiectasia mutated. These findings suggest the possible involvement of decatenation G(2) checkpoint impairment in the development of human lung cancers, as well as the potential clinical implication of selective killing of lung cancer cells with such defects by this type of topoisomerase II inhibitor.
Assuntos
Dano ao DNA , Fase G2/fisiologia , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , DNA Helicases/genética , DNA Topoisomerases Tipo II/metabolismo , DNA de Neoplasias/genética , DNA de Neoplasias/metabolismo , Dicetopiperazinas , Exodesoxirribonucleases , Fase G2/efeitos dos fármacos , Fase G2/genética , Fase G2/efeitos da radiação , Humanos , Neoplasias Pulmonares/enzimologia , Piperazinas/farmacologia , RecQ Helicases , Inibidores da Topoisomerase II , Helicase da Síndrome de WernerRESUMO
Rodents show grooming, a typical self-care behavior, under stress and non-stress conditions. Previous studies revealed that grooming under stress conditions such as the open-field test (OFT) or the elevated plus-maze test (EPM) is associated with anxiety, but the roles of grooming under non-stress conditions are not well understood. Here, we examined spray-induced grooming as a model of grooming under a non-stress condition to investigate the relationship between this grooming and depression-like behavior in the forced swim test (FST) and tail suspension test, and we compared spray-induced grooming with OFT- and EPM-induced grooming. The main finding was that the duration of spray-induced grooming, but not that of OFT/EPM-induced grooming, was negatively correlated with the duration of immobility in the FST, an index of depression-like behavior. The results suggest that spray-induced grooming is functionally different from the grooming in the OFT and EPM and is related to reduction of depressive behavior.
Assuntos
Depressão/psicologia , Asseio Animal , Animais , Modelos Animais de Doenças , Asseio Animal/fisiologia , Masculino , Aprendizagem em Labirinto , Ratos , Ratos Wistar , Estresse Psicológico/psicologia , Natação/fisiologia , Natação/psicologia , ÁguaRESUMO
In vertebrate mammals, distributed neural circuits in the brain are involved in emotion-related behavior. Netrin-G1 is a glycosyl-phosphatidylinositol-anchored synaptic adhesion molecule whose deficiency results in impaired fear-like and anxiety-like behaviors under specific circumstances. To understand the cell type and circuit specificity of these responses, we generated netrin-G1 conditional knockout mice with loss of expression in cortical excitatory neurons, inhibitory neurons, or thalamic neurons. Genetic deletion of netrin-G1 in cortical excitatory neurons resulted in altered anxiety-like behavior, but intact fear-like behavior, whereas loss of netrin-G1 in inhibitory neurons resulted in attenuated fear-like behavior, but intact anxiety-like behavior. These data indicate a remarkable double dissociation of fear-like and anxiety-like behaviors involving netrin-G1 in excitatory and inhibitory neurons, respectively. Our findings support a crucial role for netrin-G1 in dissociable neural circuits for the modulation of emotion-related behaviors, and provide genetic models for investigating the mechanisms underlying the dissociation. The results also suggest the involvement of glycosyl-phosphatidylinositol-anchored synaptic adhesion molecules in the development and pathogenesis of emotion-related behavior.
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Ansiedade/metabolismo , Comportamento Animal , Encéfalo/metabolismo , Medo , Rede Nervosa/metabolismo , Netrinas/metabolismo , Neurônios/metabolismo , Animais , Ansiedade/genética , Ansiedade/patologia , Encéfalo/patologia , Camundongos , Rede Nervosa/patologia , Netrinas/genética , Neurônios/patologiaRESUMO
Chromosomal abnormality is one of the hallmarks of neoplastic cells, and the persistent presence of chromosome instability (CIN) has been demonstrated in human cancers, including lung cancer. Recent progress in molecular and cellular biology as well as cytogenetics has shed light on the underlying mechanisms and the biological and clinical significance of chromosome abnormalities and the CIN phenotype. Chromosome abnormalities can be classified broadly into numerical (i.e., aneuploidy) and structural alterations (e.g., deletion, translocation, homogenously staining region (HSR), double minutes (DMs)). However, both alterations usually occur in the same cells, suggesting some overlap in their underlying mechanisms. Missegregation of chromosomes may result from various causes, including defects of mitotic spindle checkpoint, abnormal centrosome formation and failure of cytokinesis, while structural alterations of chromosomes may be caused especially by failure in the repair of DNA double-strand breaks (DSBs) due to the impairment of DNA damage checkpoints and/or DSB repair systems. Recent studies also suggest that telomere erosion may be involved. The consequential acquisition of the CIN phenotype would give lung cancer cells an excellent opportunity to efficiently alter their characteristics so as to be more malignant and suitable to their microenvironment, thereby gaining a selective growth advantage.
Assuntos
Aberrações Cromossômicas , Neoplasias Pulmonares/genética , Animais , Humanos , Neoplasias Pulmonares/etiologia , Neoplasias Pulmonares/fisiopatologia , CamundongosRESUMO
Most lung cancer patients are unfortunately uncurable and die because of widespread metastases, thus indicating the importance of identification of molecules with a crucial role in this process. Our previous expression profiling analysis of a highly metastatic lung cancer cell line, NCI-H460-LNM35, and its parental low metastatic line, NCI-H460-N15, revealed significant up-regulation of both known and unknown genes in LNM35. In this study, we describe the isolation and detailed characterizations of a novel gene, named CLCP1, which corresponds to one of such expression sequence tags with up-regulated expression in LNM35. The CLCP1 gene was found to encode a protein with 775 amino acids with structural similarities to, but distinct from neuropilins, cell surface receptors for VEGF165 and semaphorins. Notably, CLCP1 was shown to be up-regulated not only in LNM35 in association with its acquisition of metastatic phenotype during in vivo selection, but also in a significant fraction of lung cancers in vivo with high frequency in metastatic lesions, warranting future studies for a better understanding of the molecular mechanisms of lung cancer metastasis.
Assuntos
Adenocarcinoma/genética , Carcinoma de Células Grandes/genética , Carcinoma de Células Pequenas/genética , Carcinoma de Células Escamosas/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares/genética , Proteínas de Membrana/genética , Proteínas de Neoplasias/genética , Regulação para Cima , Adenocarcinoma/patologia , Adulto , Sequência de Aminoácidos , Sequência de Bases , Carcinoma de Células Grandes/patologia , Carcinoma de Células Pequenas/patologia , Carcinoma de Células Escamosas/patologia , Fracionamento Químico , Clonagem Molecular , DNA Complementar , Humanos , Neoplasias Pulmonares/patologia , Dados de Sequência Molecular , Metástase Neoplásica , Homologia de Sequência do Ácido Nucleico , Transfecção , Células Tumorais CultivadasRESUMO
BACKGROUND: Guanine nucleotide exchange factors (GEFs) activate small GTPases that are involved in several cellular functions. cAMP-guanine nucleotide exchange factor II (cAMP-GEF II) acts as a target for cAMP independently of protein kinase A (PKA) and functions as a GEF for Rap1 and Rap2. Although cAMP-GEF II is expressed abundantly in several brain areas including the cortex, striatum, and hippocampus, its specific function and possible role in hippocampal synaptic plasticity and cognitive processes remain elusive. Here, we investigated how cAMP-GEF II affects synaptic function and animal behavior using cAMP-GEF II knockout mice. RESULTS: We found that deletion of cAMP-GEF II induced moderate decrease in long-term potentiation, although this decrease was not statistically significant. On the other hand, it produced a significant and clear impairment in NMDA receptor-dependent long-term depression at the Schaffer collateral-CA1 synapses of hippocampus, while microscopic morphology, basal synaptic transmission, and depotentiation were normal. Behavioral testing using the Morris water maze and automated IntelliCage system showed that cAMP-GEF II deficient mice had moderately reduced behavioral flexibility in spatial learning and memory. CONCLUSIONS: We concluded that cAMP-GEF II plays a key role in hippocampal functions including behavioral flexibility in reversal learning and in mechanisms underlying induction of long-term depression.
Assuntos
Comportamento Animal , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Hipocampo/metabolismo , Potenciação de Longa Duração , Animais , Encéfalo/metabolismo , Eletrochoque , Fatores de Troca do Nucleotídeo Guanina/deficiência , Aprendizagem , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores de N-Metil-D-Aspartato/metabolismo , Sinapses/metabolismoRESUMO
ZD1839 ('Iressa') is an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) that inhibits EGFR signaling. Emerging evidence indicates that ZD1839 has clinical potential in lung cancer, but very little is known about the molecular characteristics of lung cancers that may determine sensitivity to ZD1839. We examined a panel of 19 lung cancer cell lines to investigate possible association between ZD1839 sensitivity and histological type, expression level and constitutive phosphorylation of EGFR and K-ras gene status. Our results indicate that neither expression level nor constitutive activation status of EGFR seems to predict sensitivity to ZD1839. In addition, ZD1839 sensitivity was not associated with expression of human epidermal growth factor receptor-2 (HER-2), another member of this tyrosine kinase receptor family nor with co-expression of EGFR and HER-2. Finally, no correlation was found between the presence of activating mutations of the K-ras gene, an important downstream mediator of the EGFR-transduced signals and the relative resistance to ZD1839. These findings warrant future study to clarify how ZD1839 inhibits lung cancer cell growth and to find a useful marker for prediction of sensitivity to this novel and promising agent for the treatment of lung cancers.
Assuntos
Inibidores Enzimáticos/uso terapêutico , Receptores ErbB/metabolismo , Genes ras/fisiologia , Neoplasias Pulmonares/tratamento farmacológico , Quinazolinas/uso terapêutico , Receptor ErbB-2/metabolismo , Antineoplásicos/uso terapêutico , Resistencia a Medicamentos Antineoplásicos , Fator de Crescimento Epidérmico/antagonistas & inibidores , Gefitinibe , Humanos , Neoplasias Pulmonares/metabolismo , Fosforilação/efeitos dos fármacos , Proteínas Tirosina Quinases/antagonistas & inibidores , Células Tumorais CultivadasRESUMO
The effects of phenylthiourea (PTU) and its analogues on chick embryonic pigmented epithelial cells (PECs) in culture were studied to elucidate the correlation between inhibition of melanogenesis of PECs and enhancement of their transdifferentiation into lens cells. Both 0.25-0.5 mM PTU and 0.1 mM alpha-naphthylthiourea (ANTU) effectively inhibited melanogenesis of PECs and stimulated their transdifferentiation into lens cells at the same time. Thiourea (TU) also inhibited melanogenesis at a much higher concentration (4 mM), but did not stimulate the lens transdifferentiation at all. Methylthiourea (MTU), on the other hand, did not inhibit melanogenesis, but stimulated the lens transdifferentiation. Testicular hyaluronidase effectively amplified the above-mentioned stimulating effects of thioureas without their altering optimum concentrations, although this enzyme itself never enhanced the lens transdifferentiation of PECs but suppressed their melanogenesis at a concentration of 100 U/ml medium, onward. These results suggest that the suppression of melanogenesis of PECs by PTU or its analogues does not directly correlate with their transdifferentiation into lens cells. The possible mode of thiourea actions on the lens transdifferentiation of PECs cultured in vitro is discussed.
RESUMO
In humans, exposure to high levels of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is associated with chronic obstructive pulmonary disease and lung cancer. While several studies have shown that the lung is a target organ for TCDD toxicity, little is known on the specific biological pathways altered by TCDD. Studies have shown that the transcriptional response of TCDD (in vivo and in vitro) is complex, and exhibits cell type and tissue specificity. Thus, the purpose of this study was to look at global and concentration-dependent effects of TCDD on gene expression in human lung cells. Gene expression profiling of both a nontumorigenic (HPL1A) and a malignant, tumorigenic lung cell line (A549) was performed by microarray dual fluorescence hybridizations in cells treated with increasing concentrations of TCDD (0, 0.1, 1, 10 nM) for 24 h. Real time RT-PCR was used to verify alterations in specific genes. Results showed that 68 out of 2091 genes were changed in each cell line, and 15 of those genes were found altered in both cell lines. Common gene responses altered by TCDD were identified and included known xenobiotic metabolizing genes, genes known to alter cell cycle, as well as genes that are involved with cell signaling and that mediate cell motility or communication. Cell line specific differences in gene expression were found that indicate the nonmalignant HPL1A cells are retinoic acid responsive. In addition, TCDD altered specific immunomodulatory genes in the HPL1A cells. These data show that TCDD alters multiple integrated networks of signaling pathways associated with pulmonary disease, particularly that of lung cancer.