RESUMO
Meningeal carcinomatosis (MC) has an extremely poor prognosis and can present with various neurological symptoms. A 68-year-old man presented to our hospital with a 1 month history of headache and nausea followed by sudden decrease in vision in both eyes. Whilst being examined in the ophthalmology department he lost consciousness and had a generalised tonic clonic seizure. Accordingly, he was transferred to the Emergency Department. Head magnetic resonance imaging showed hydrocephalus. Abdominal contrast-enhanced computed tomography scanning reported changes suggestive of gastric carcinoma. Cerebrospinal fluid cytological examination showed numerous atypical cells, leading to the diagnosis of MC. An upper gastrointestinal endoscopy revealed an advanced gastric tumour. Systemic chemotherapy was initiated, however, he died within 16 days of admission. At autopsy, poorly differentiated adenocarcinoma was identified in the subarachnoid space, however it had not invaded the brain parenchyma or optic chiasm. This is the first report of loss of vision being the first presenting symptom of new-onset gastric carcinoma with MC. Although rare, MC should be suspected in cases where patients present with sudden loss of vision and symptoms of meningeal irritation, where there are no ophthalmological findings to explain the vision loss.
RESUMO
Mutations of the leucine-rich glioma-inactivated 1 (LGI1) gene cause an autosomal dominant partial epilepsy with auditory features also known as autosomal-dominant lateral temporal lobe epilepsy. LGI1 is also the main antigen present in sera and cerebrospinal fluids of patients with limbic encephalitis and seizures, highlighting its importance in a spectrum of epileptic disorders. LGI1 encodes a neuronal secreted protein, whose brain function is still poorly understood. Here, we generated, by ENU (N-ethyl-N-nitrosourea) mutagenesis, Lgi1-mutant rats carrying a missense mutation (L385R). We found that the L385R mutation prevents the secretion of Lgi1 protein by COS7 transfected cells. However, the L385R-Lgi1 protein was found at low levels in the brains and cultured neurons of Lgi1-mutant rats, suggesting that mutant protein may be destabilized in vivo. Studies on the behavioral phenotype and intracranial electroencephalographic signals from Lgi1-mutant rats recalled several features of the human genetic disorder. We show that homozygous Lgi1-mutant rats (Lgi1(L385R/L385R)) generated early-onset spontaneous epileptic seizures from P10 and died prematurely. Heterozygous Lgi1-mutant rats (Lgi1(+/L385R)) were more susceptible to sound-induced, generalized tonic-clonic seizures than control rats. Audiogenic seizures were suppressed by antiepileptic drugs such as carbamazepine, phenytoin and levetiracetam, which are commonly used to treat partial seizures, but not by the prototypic absence seizure drug, ethosuximide. Our findings provide the first rat model with a missense mutation in Lgi1 gene, an original model complementary to knockout mice. This study revealed that LGI1 disease-causing missense mutations might cause a depletion of the protein in neurons, and not only a failure of Lgi1 secretion.
Assuntos
Epilepsia/etiologia , Proteínas/genética , Proteínas/metabolismo , Sequência de Aminoácidos , Animais , Anticonvulsivantes/farmacologia , Encéfalo/metabolismo , Células COS , Carbamazepina/farmacologia , Células Cultivadas , Chlorocebus aethiops , Modelos Animais de Doenças , Eletroencefalografia , Epilepsias Parciais/tratamento farmacológico , Epilepsias Parciais/genética , Epilepsia/tratamento farmacológico , Epilepsia/genética , Epilepsia Reflexa/tratamento farmacológico , Epilepsia Reflexa/genética , Etossuximida/farmacologia , Heterozigoto , Homozigoto , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Levetiracetam , Dados de Sequência Molecular , Mutação de Sentido Incorreto , Neurônios/metabolismo , Fenitoína/farmacologia , Piracetam/análogos & derivados , Piracetam/farmacologia , Ratos MutantesRESUMO
The dorsal striatum, which contains the dorsolateral striatum (DLS) and dorsomedial striatum (DMS), integrates the acquisition and implementation of instrumental learning in cooperation with the nucleus accumbens (NAc). The dorsal striatum regulates the basal ganglia circuitry through direct and indirect pathways. The mechanism by which these pathways mediate the learning processes of instrumental actions remains unclear. We investigated how the striatal indirect (striatopallidal) pathway arising from the DLS contributes to the performance of conditional discrimination. Immunotoxin targeting of the striatal neuronal type containing dopamine D(2) receptor in the DLS of transgenic rats resulted in selective, efficient elimination of the striatopallidal pathway. This elimination impaired the accuracy of response selection in a two-choice reaction time task dependent on different auditory stimuli. The impaired response selection was elicited early in the test sessions and was gradually restored as the sessions continued. The restoration from the deficits in auditory discrimination was prevented by excitotoxic lesion of the NAc but not by that of the DMS. In addition, lesion of the DLS mimicked the behavioral consequence of the striatopallidal removal at the early stage of test sessions of discriminative performance. Our results demonstrate that the DLS-derived striatopallidal pathway plays an essential role in the execution of conditional discrimination, showing its contribution to the control of selection accuracy of learned motor responses. The results also suggest the presence of a mechanism that compensates for the learning deficits during the repetitive sessions, at least partly, demanding accumbal function.
Assuntos
Condicionamento Operante/fisiologia , Corpo Estriado/fisiologia , Discriminação Psicológica/fisiologia , Atividade Motora/fisiologia , Estimulação Acústica , Análise de Variância , Animais , Animais Geneticamente Modificados , Biotina/análogos & derivados , Calbindina 2 , Comportamento de Escolha/efeitos dos fármacos , Comportamento de Escolha/fisiologia , Colina O-Acetiltransferase/metabolismo , Condicionamento Operante/efeitos dos fármacos , Corpo Estriado/citologia , Corpo Estriado/lesões , Dextranos , Neurônios Dopaminérgicos/efeitos dos fármacos , Encefalinas/genética , Encefalinas/metabolismo , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/genética , Ácido Ibotênico/toxicidade , Imunotoxinas/toxicidade , Interneurônios/metabolismo , Masculino , Motivação/efeitos dos fármacos , Motivação/genética , Parvalbuminas/metabolismo , Fosfopiruvato Hidratase/metabolismo , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , Ratos , Ratos Long-Evans , Tempo de Reação/efeitos dos fármacos , Tempo de Reação/genética , Receptores de Dopamina D2/deficiência , Receptores de Dopamina D2/metabolismo , Receptores de Interleucina-2/genética , Esquema de Reforço , Proteína G de Ligação ao Cálcio S100/metabolismo , Substância Negra/metabolismo , Taquicininas/genética , Taquicininas/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo , Área Tegmentar Ventral/metabolismoRESUMO
To establish low density lipoprotein receptor (LDLR) mutant rats as a hypercholesterolemia and atherosclerosis model, we screened the rat LDLR gene for mutations using an N-ethyl-N-nitrosourea mutagenesis archive of rat gene data, and identified five mutations in its introns and one missense mutation (478T>A) in exon 4. The C160S mutation was located in the ligand binding domain of LDLR and was revealed to be equivalent to mutations (C160Y/G) identified in human familial hypercholesterolemia (FH) patients. The wild type, heterozygous, and homozygous mutant rats were fed a normal chow diet or a high fat high cholesterol (HFHC) diet from the age of 10 weeks for 16 weeks. The LDLR homozygous mutants fed the normal chow diet showed higher levels of plasma total cholesterol and LDL cholesterol than the wild type rats. When fed the HFHC diet, the homozygous mutant rats exhibited severe hyperlipidemia and significant lipid deposition from the aortic arch to the abdominal aorta as well as in the aortic valves. Furthermore, the female homozygous mutants also developed xanthomatosis in their paws. In conclusion, we suggest that LDLR mutant rats are a useful novel animal model of hypercholesterolemia and atherosclerosis.
Assuntos
Aterosclerose/genética , Hipercolesterolemia/genética , Receptores de LDL/genética , Animais , Aterosclerose/sangue , Aterosclerose/patologia , Modelos Animais de Doenças , Feminino , Hipercolesterolemia/sangue , Hipercolesterolemia/patologia , Lipídeos/sangue , Masculino , Mutação , Ratos , Ratos Endogâmicos F344 , Ratos MutantesRESUMO
OBJECTIVE: To evaluate the sensitivity and specificity of endometrial cytology obtained by intrauterine sample using a descriptive reporting format for endometrial cytological diagnosis. STUDY DESIGN: 10,152 consecutive endometrial scrapings obtained in 13 different Japanese hospitals were analyzed. Cytological results were classified as 'negative for malignancy', 'atypical endometrial cells' (ATEC), 'endometrial hyperplasia', 'atypical endometrial hyperplasia' or 'malignant tumor'. ATEC was subclassified as 'ATEC, of undetermined significance' (ATEC-US) and 'ATEC, cannot exclude atypical endometrial hyperplasia or more' (ATEC-A). Cytological results were compared with the histological diagnosis as a gold standard. When the cytological result was 'negative for malignancy' and there was no subsequent histological examination, the case was considered a true negative when the endometrium was assessed as normal on transvaginal ultrasonography and there was no abnormal uterine bleeding. RESULTS: 1,083 cases in which histology was not performed, 557 cases of 'unsatisfactory specimen' and 76 cases of ATEC-US were excluded. In the remaining 8,436 cases, the sensitivity and specificity, positive predictive value and negative predictive value for detecting atypical endometrial hyperplasia or malignant tumors were 79.0 and 99.7, 92.9 and 98.9%, respectively. CONCLUSION: The current diagnostic standards for endometrial cytology in Japan were established. Specificity is satisfactory for excluding cancer or precancerous diseases.
Assuntos
Neoplasias do Endométrio/classificação , Neoplasias do Endométrio/patologia , Endométrio/patologia , Patologia Clínica/normas , Sociedades Médicas/normas , Terminologia como Assunto , Adenocarcinoma/classificação , Adenocarcinoma/patologia , Citodiagnóstico/métodos , Citodiagnóstico/normas , Neoplasias do Endométrio/diagnóstico por imagem , Endométrio/diagnóstico por imagem , Feminino , Humanos , Japão , Patologia Clínica/métodos , Ultrassonografia , Neoplasias do Colo do Útero/classificação , Neoplasias do Colo do Útero/patologiaRESUMO
In vitro fertilization (IVF) is a valuable technique for the propagation of experimental animals. IVF has typically been used in mice to rapidly expand breeding colonies and create large numbers of embryos. However, applications of IVF in rat breeding experiments have stalled due to the inconvenient laboratory work schedules imposed by current IVF protocols for this species. Here, we developed a new rat IVF protocol that consists of experimental steps performed during common laboratory working hours. Our protocol can be completed within 12 h by shortening the period of sperm capacitation from 5 to 1 h and the fertilization time from 10 to 8 h in human tubal fluid (HTF) medium. This new protocol generated an excellent birth rate and was applicable not only to closed colony rat strains, such as Wistar, Long-Evans, and Sprague-Dawley (SD), but also to the inbred Lewis strain. Moreover, Wistar and Long-Evans embryos prepared by this protocol were successfully frozen by vitrification and later successfully thawed and resuscitated. This protocol is practical and can be easily adopted by laboratory workers.
Assuntos
Embrião de Mamíferos/fisiologia , Fertilização in vitro/métodos , Ciência dos Animais de Laboratório/métodos , Animais , Cruzamento/métodos , Criopreservação/métodos , Meios de Cultura/química , Tubas Uterinas/fisiologia , Feminino , Masculino , Oócitos/fisiologia , Ratos , Ratos Endogâmicos Lew , Ratos Long-Evans , Ratos Sprague-Dawley , Ratos Wistar , Ressuscitação/métodos , Especificidade da Espécie , Espermatozoides/fisiologia , Fatores de Tempo , VitrificaçãoRESUMO
Previous studies have demonstrated that mutation in the forkhead domain of the forkhead box P2 (FOXP2) protein (R553H) causes speech-language disorders. To further analyze FOXP2 function in speech learning, we generated a knockin (KI) mouse for Foxp2 (R552H) [Foxp2 (R552H)-KI], corresponding to the human FOXP2 (R553H) mutation, by homologous recombination. Homozygous Foxp2 (R552H)-KI mice showed reduced weight, immature development of the cerebellum with incompletely folded folia, Purkinje cells with poor dendritic arbors and less synaptophysin immunoreactivity, and achieved crisis stage for survival 3 weeks after birth. At postnatal day 10, these mice also showed severe ultrasonic vocalization (USV) and motor impairment, whereas the heterozygous Foxp2 (R552H)-KI mice exhibited modest impairments. Similar to the wild-type protein, Foxp2 (R552H) localized in the nuclei of the Purkinje cells and the thalamus, striatum, cortex, and hippocampus (CA1) neurons of the homozygous Foxp2 (R552H)-KI mice (postnatal day 10), and some of the neurons showed nuclear aggregates of Foxp2 (R552H). In addition to the immature development of the cerebellum, Foxp2 (R552H) nuclear aggregates may further compromise the function of the Purkinje cells and cerebral neurons of the homozygous mice, resulting in their death. In contrast, heterozygous Foxp2 (R552H)-KI mice, which showed modest impairment of USVs with different USV qualities and which did not exhibit nuclear aggregates, should provide insights into the common molecular mechanisms between the mouse USV and human speech learning and the relationship between the USV and motor neural systems.
Assuntos
Fatores de Transcrição Forkhead/genética , Transtornos da Linguagem/genética , Células de Purkinje/citologia , Proteínas Repressoras/genética , Vocalização Animal , Animais , Cerebelo/crescimento & desenvolvimento , Primers do DNA/genética , Genótipo , Histocitoquímica , Camundongos , Camundongos Transgênicos , Análise em Microsséries , Mutação de Sentido Incorreto/genética , UltrassomRESUMO
OBJECTIVE: To investigate the biological significance of single nucleotide polymorphism (SNP) at murine double-minute 2 homolog (MDM2) promoter 309 in cervical carcinogenesis. METHODS: SNP at MDM2 promoter 309 (T/G) together with human papillomavirus (HPV) types was examined in a total of 195 cervical smear samples and 8 human cervical squamous carcinoma cell lines using two independent PCR assays and PCR-RFLP techniques. RESULTS: Forty-one patients with high-grade squamous intraepithelial lesion (HSIL) had higher frequency of high-risk HPV than 102 with low-grade SIL (LSIL) and 52 controls. There was an increased OR (8.88; CI=2.34-33.63; P=0.003) for TG+GG genotype in HSIL cases compared to controls among 68 patients with high-risk HPV. Twenty-one cases with HPV types 16 and/or 18 had significantly higher frequency of the TG+GG genotype and G allele than 47 with other types of high-risk HPV. Seven of 8 cervical carcinoma cell lines also showed TG or GG genotype. CONCLUSION: MDM2-SNP309 (T/G) and high-risk HPV infection may be closely associated with cervical carcinogenesis in a Japanese population.
Assuntos
Transformação Celular Viral/genética , Papillomaviridae/genética , Infecções por Papillomavirus/genética , Proteínas Proto-Oncogênicas c-mdm2/genética , Neoplasias do Colo do Útero/genética , Alelos , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/virologia , Linhagem Celular Tumoral , Feminino , Humanos , Infecções por Papillomavirus/patologia , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Regiões Promotoras Genéticas , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologia , Displasia do Colo do Útero/genética , Displasia do Colo do Útero/patologia , Displasia do Colo do Útero/virologiaRESUMO
BACKGROUND: Establishing a peritoneal mesothelial cell (MC) line in which the native characteristics of primary MCs are constantly maintained in vivo is of great significance for investigating their morphological and functional changes in peritoneal dialysis. We established transgenic (Tg) rats that expressed the temperature-sensitive tsA58 mutant of the simian virus 40 large T-antigen (tsSV40T), which served as a source of immortalized rat cell lines. The cells were immortalized at a permissive temperature of 33 degrees C, although they were differentiated at a non-permissive temperature of 38 degrees C. In this study, we established a novel MC line from tsSV40T Tg rats and evaluated its characteristics. METHODS: MCs were isolated from 8-week-old tsSV40T Tg rats and cloned. MCs from 8-week-old Wistar rats were used as controls. These cells were immunohistochemically and phenotypically evaluated by immunofluorescence, phase contrast and electron microscopy. The production of plasminogen activator inhibitor 1 (PAI-1) from MCs stimulated by tumour necrosis factor-alpha (TNF-alpha) was measured. RESULTS: The tsSV40T MCs showed a cobblestone-like appearance at 33 and 38 degrees C, which was similar to normal primary cultured MCs. Microvilli-like structures were observed on the cell surface by a scanning electron microscope at 33 and 38 degrees C. Wilms tumour-1 and pancytokeratin, as MC markers, were expressed at 33 and 38 degrees C. Following TNF-alpha stimulation, PAI-1 production of tsSV40T MCs was similar to that of normal primary cultured MCs. CONCLUSION: We established a novel, conditionally immortalized MC line that continuously retained the characteristics of primary cultured peritoneal MCs. This cell line might be a useful tool for various types of in vitro biological research on peritoneal dialysis.
Assuntos
Antígenos Transformantes de Poliomavirus/genética , Peritônio/citologia , Vírus 40 dos Símios/genética , Vírus 40 dos Símios/imunologia , Animais , Animais Geneticamente Modificados , Sequência de Bases , Linhagem Celular , Proliferação de Células , Transformação Celular Viral/genética , Transformação Celular Viral/imunologia , DNA/biossíntese , Primers do DNA/genética , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Expressão Gênica , Genes Virais , Mutação , Peritônio/metabolismo , Inibidor 1 de Ativador de Plasminogênio/biossíntese , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Viral/genética , RNA Viral/metabolismo , Ratos , Temperatura , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
Clinical practice guidelines for gynecologic cancers have been published by the National Comprehensive Cancer Network and the National Cancer Institute. Whereas these guidelines form the basis for the standard of care for gynecologic malignancies in the United States, it has proven difficult to institute them in Japan due to differences in patient characteristics, health-care delivery systems, and insurance programs. Therefore, evidence-based guidelines for treating cervical cancer specifically in Japan have been under development. The Guidelines Formulation Committee and Evaluation Committee were independently established within the Committee for Treatment Guidelines for Cervical Cancer. Opinions from within and outside the Japan Society of Gynecologic Oncology (JSGO) were incorporated into the final draft, and the guidelines were published after approval by the JSGO. These guidelines are composed of ten chapters and comprise three algorithms. Each chapter consists of a clinical question, recommendations, background, objectives, explanations, and references. The objective of these guidelines is to clearly delineate the standard of care for cervical cancer treatment in Japan in order to ensure equitable care for all Japanese women diagnosed with cervical cancer.
Assuntos
Algoritmos , Medicina Baseada em Evidências , Seleção de Pacientes , Neoplasias do Colo do Útero/terapia , Árvores de Decisões , Feminino , Humanos , Japão , Estadiamento de Neoplasias , Sociedades Médicas , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/patologiaRESUMO
BACKGROUND: Cervical cancer is caused primarily by human papillomaviruses (HPV). The polymorphism rs1042522 at codon 72 of the TP53 tumour-suppressor gene has been investigated as a genetic cofactor. More than 80 studies were done between 1998 and 2006, after it was initially reported that women who are homozygous for the arginine allele had a risk for cervical cancer seven times higher than women who were heterozygous for the allele. However, results have been inconsistent. Here we analyse pooled data from 49 studies to determine whether there is an association between TP53 codon 72 polymorphism and cervical cancer. METHODS: Individual data on 7946 cases and 7888 controls from 49 different studies worldwide were reanalysed. Odds ratios (OR) were estimated using logistic regression, stratifying by study and ethnic origin. Subgroup analyses were done for infection with HPV, ethnic origin, Hardy-Weinberg equilibrium, study quality, and the material used to determine TP53 genotype. FINDINGS: The pooled estimates (OR) for invasive cervical cancer were 1.22 (95% CI 1.08-1.39) for arginine homozygotes compared with heterozygotes, and 1.13 (0.94-1.35) for arginine homozygotes versus proline homozygotes. Subgroup analyses showed significant excess risks only in studies where controls were not in Hardy-Weinberg equilibrium (1.71 [1.21-2.42] for arginine homozygotes compared with heterozygotes), in non-epidemiological studies (1.35 [1.15-1.58] for arginine homozygotes compared with heterozygotes), and in studies where TP53 genotype was determined from tumour tissue (1.39 [1.13-1.73] for arginine homozygotes compared with heterozygotes). Null results were noted in studies with sound epidemiological design and conduct (1.06 [0.87-1.29] for arginine homozygotes compared with heterozygotes), and studies in which TP53 genotype was determined from white blood cells (1.06 [0.87-1.29] for arginine homozygotes compared with heterozygotes). INTERPRETATION: Subgroup analyses indicated that excess risks were most likely not due to clinical or biological factors, but to errors in study methods. No association was found between cervical cancer and TP53 codon 72 polymorphism when the analysis was restricted to methodologically sound studies. FUNDING: German Research Foundation (DFG).
Assuntos
Genes p53 , Predisposição Genética para Doença , Polimorfismo Genético , Neoplasias do Colo do Útero/genética , Adolescente , Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/genética , Neoplasias do Colo do Útero/virologia , Adulto JovemRESUMO
Patients with poorly differentiated endometrial cancer show poor prognosis, and effective molecular target-based therapies are needed. Endometrial cancer cells proliferate depending on the activation of HES1 (hairy and enhancer of split-1), which is induced by several pathways, such as the Notch and fibroblast growth factor receptor (FGFR) signaling pathways. In addition, aberrant, ligand-free activation of the FGFR signaling pathway resulting from mutations in FGFR2 was also reported in endometrial cancer. However, a clinical trial showed that there was no difference in the effectiveness of FGFR inhibitors between patients with and without the FGFR2 mutation, suggesting a presence of another signaling pathway for the FGFR activation. Here, we investigated the signaling pathway regulating the expression of HES1 and proliferation of poorly and well-differentiated endometrial cancer cell lines Ishikawa and HEC-50B, respectively. Whereas Ishikawa cells proliferated and expressed HES1 in a Notch signaling-dependent manner, Notch signaling was not involved in HES1 and proliferation of HEC-50B cells. The FGFR inhibitor, NVP-BGJ398, decreased HES1 expression and proliferation of HEC-50B cells; however, HEC50B cells had no mutations in the FGFR2 gene. Instead, HEC-50B cells highly expressed ligands for FGFR2, suggesting that FGFR2 is activated by an autocrine manner, not by ligand-free activation. This autocrine pathway activated Akt downstream of FGFR for cell proliferation. Our findings suggest the usefulness of HES1 as a marker for the proliferation signaling and that FGFR inhibitor may be effective for poorly differentiated endometrial cancers that harbor wild-type FGFR.
Assuntos
Comunicação Autócrina/genética , Proliferação de Células/genética , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/patologia , Expressão Gênica , Receptores de Fatores de Crescimento de Fibroblastos/genética , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo , Fatores de Transcrição HES-1/genética , Fatores de Transcrição HES-1/metabolismo , Linhagem Celular Tumoral , Neoplasias do Endométrio/terapia , Feminino , Humanos , Terapia de Alvo Molecular , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/metabolismo , Receptores Notch/metabolismo , Transdução de SinaisRESUMO
The objective of this study was to establish pure blood-nerve barrier (BNB)-derived peripheral nerve pericyte cell lines and to investigate their unique properties as barrier-forming cells. We isolated peripheral nerve, brain, and lung pericytes from transgenic rats harboring the temperature-sensitive simian virus 40 large T-antigen gene. These cell lines expressed several pericyte markers such as alpha-smooth muscle actin, NG2, osteopontin, and desmin, whereas they did not express endothelial cell markers such as vWF and PECAM. In addition, these cell lines expressed several tight junction molecules such as occludin, claudin-12, ZO-1, and ZO-2. In particular, the expression of occludin was detected in peripheral nerve and brain pericytes, although it was not detected in lung pericytes by a Western blot analysis. An immunocytochemical analysis confirmed that occludin and ZO-1 were localized at the cell-cell boundaries among the pericytes. Brain and peripheral nerve pericytes also showed significantly higher trans-pericyte electrical resistance values and lower inulin clearances than lung pericytes. We considered that occludin localized at the cell-cell boundaries among the pericytes might mechanically stabilize the microvessels of the BNB and the blood-brain barrier. Furthermore, we also showed that these cell lines expressed many barrier-related transporters. ABCG2, p-gp, MRP-1, and Glut-1 were detected by a Western blot analysis and were observed in the cytoplasm and outer membrane by an immunocytochemical analysis. These transporters on pericytes might facilitate the peripheral nerve-to-blood efflux and blood-to-peripheral nerve influx transport of substrates in cooperation with those on endothelial cells in order to maintain peripheral nerve homeostasis.
Assuntos
Barreira Hematoneural/metabolismo , Permeabilidade Capilar , Proteínas de Membrana/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Pericitos/metabolismo , Nervo Isquiático/irrigação sanguínea , Junções Íntimas/metabolismo , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Animais , Animais Geneticamente Modificados , Antígenos Transformantes de Poliomavirus/genética , Antígenos Transformantes de Poliomavirus/metabolismo , Apoptose , Barreira Hematoneural/citologia , Western Blotting , Encéfalo/irrigação sanguínea , Linhagem Celular , Resistência a Medicamentos , Impedância Elétrica , Feminino , Imuno-Histoquímica , Inulina/metabolismo , Pulmão/irrigação sanguínea , Proteínas de Membrana/genética , Proteínas de Membrana Transportadoras/genética , Pericitos/efeitos dos fármacos , Puromicina/farmacologia , RNA Mensageiro/metabolismo , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Regulação para CimaRESUMO
Reproductive isolation that initiates speciation is likely caused by incompatibility among multiple loci in organisms belonging to genetically diverging populations. Laboratory C57BL/6J mice, which predominantly originated from Mus musculus domesticus, and a MSM/Ms strain derived from Japanese wild mice (M. m. molossinus, genetically close to M. m. musculus) are reproductively isolated. Their F1 hybrids are fertile, but successive intercrosses result in sterility. A consomic strain, C57BL/6J-ChrX(MSM), which carries the X chromosome of MSM/Ms in the C57BL/6J background, shows male sterility, suggesting a genetic incompatibility of the MSM/Ms X chromosome and other C57BL/6J chromosome(s). In this study, we conducted genomewide linkage analysis and subsequent QTL analysis using the sperm shape anomaly that is the major cause of the sterility of the C57BL/6J-ChrX(MSM) males. These analyses successfully detected significant QTL on chromosomes 1 and 11 that interact with the X chromosome. The introduction of MSM/Ms chromosomes 1 and 11 into the C57BL/6J-ChrX(MSM) background failed to restore the sperm-head shape, but did partially restore fertility. This result suggests that this genetic interaction may play a crucial role in the reproductive isolation between the two strains. A detailed analysis of the male sterility by intracytoplasmic sperm injection and zona-free in vitro fertilization demonstrated that the C57BL/6J-ChrX(MSM) spermatozoa have a defect in penetration through the zona pellucida of eggs.
Assuntos
Hibridização Genética , Camundongos/classificação , Reprodução , Testículo/fisiologia , Cromossomo X , Animais , Cruzamentos Genéticos , Feminino , Fertilização in vitro , Ligação Genética , Genótipo , Endogamia , Masculino , Camundongos/genética , Camundongos Endogâmicos C57BL , Locos de Características Quantitativas , Cromossomo Y/genéticaRESUMO
A tracheal epithelial cell line RTEC11 was established from transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen. The cells grew continuously at a permissive temperature of 33 degrees C but not at a non-permissive temperature of 39 degrees C. Morphological and functional investigations demonstrated that the cells were polarized epithelial cells maintaining a regulated permeability barrier function. Interestingly, the expression levels of Muc1 (mucin 1) and Scgb1a1 (uteroglobin), non-ciliated secretory cell markers, and Tubb4 (tubulin beta 4), a ciliated cell marker, were significantly increased under the cell growth-restricted condition. Global gene expression and computational network analyses demonstrated a significant genetic network associated with cellular development and differentiation in cells cultured at the non-permissive temperature. The tracheal epithelial cell line RTEC11 with unique characteristics should be useful as an in vitro model for studies of the physiological functions and gene expression of tracheal epithelial cells.
Assuntos
Antígenos Transformantes de Poliomavirus/genética , Antígenos Transformantes de Poliomavirus/metabolismo , Células Epiteliais/metabolismo , Temperatura , Traqueia/metabolismo , Animais , Biomarcadores/análise , Biomarcadores/metabolismo , Proteínas de Transporte/metabolismo , Técnicas de Cultura de Células , Diferenciação Celular/genética , Linhagem Celular , Membrana Celular/química , Membrana Celular/metabolismo , Permeabilidade da Membrana Celular/genética , Proliferação de Células , Cílios/metabolismo , Simulação por Computador , Células Epiteliais/citologia , Regulação da Expressão Gênica/genética , Modelos Biológicos , Mucina-1/metabolismo , Redes Neurais de Computação , Ratos , Ratos Transgênicos , Traqueia/citologia , Tubulina (Proteína)/metabolismo , UteroglobinaRESUMO
To better understand the molecular mechanisms of carcinogenesis induced in uterine endometrium by therapeutic anti-estrogenic Tamoxifen (TAM) exposure, 27 uterine tumors (4 benign endometrial polyps and 23 carcinomas) associated with TAM exposure were analyzed for the presence and spectrum of p53 and K-ras mutations. Although there was no significant difference between TAM-associated endometrial carcinomas and sporadic endometrial tumors in the frequency of these mutations, the spectrum of p53 mutations was characteristically unique to the TAM-associated tumors. The median duration of TAM exposure was significantly longer in patients with p53 mutations than those without p53 mutations (62 vs. 30 months, p=0.028). Our observation suggests that prolonged TAM exposure may directly inactivate the p53 gene by acting as a mutagen in a significant fraction of TAM-associated endometrial carcinomas.
Assuntos
Antineoplásicos Hormonais/efeitos adversos , Carcinoma/induzido quimicamente , Carcinoma/metabolismo , Neoplasias do Endométrio/induzido quimicamente , Neoplasias do Endométrio/metabolismo , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Genes p53 , Genes ras , Mutação , Tamoxifeno/efeitos adversos , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
Genes of the RAF family, which mediate cellular responses to growth signals, encode kinases that are regulated by RAS and participate in the RAS, RAF, mitogen/extracellular signal-regulated kinase, extracellular signal-regulated kinase and mitogen-activated protein kinase pathway. As BRAF is a serine/threonine kinase that is commonly activated by somatic point mutation, it may provide possible diagnostic and therapeutic targets in human malignant tumors. We analyzed exon 15 of the BRAF gene for mutations in 58 lung, 12 breast, six kidney, 14 cervical, four endometrial and 10 ovarian carcinoma cell lines by PCR-SSCP and direct sequencing. The T1796A transversion was found in one (2.9%) of 34 small cell lung carcinoma and one (8.3%) of 12 breast carcinoma cell lines, resulting in a valine-to-glutamate substitution at residue 599 (V599E). One (4.2%) of 24 non-small cell lung carcinoma cell line showed the C1786G transversion, leading to a leucine-to-valine substitution at residue 596 (L596V). No BRAF point mutations were found in any of the other cell lines examined. Our present results suggest that BRAF may not be a frequent target of mutations involved in the pathogenesis of human lung, breast, kidney, cervical, endometrial and ovarian carcinomas.
Assuntos
Neoplasias/genética , Mutação Puntual/genética , Proteínas Proto-Oncogênicas B-raf/genética , Substituição de Aminoácidos/genética , Linhagem Celular Tumoral , Análise Mutacional de DNA/métodos , Éxons/genética , Humanos , Reação em Cadeia da Polimerase/métodos , Polimorfismo Conformacional de Fita SimplesRESUMO
The multifactorial process of carcinogenesis involves mutations in oncogenes, or tumor suppressor genes, as well as the influence of environmental etiological factors. Common DNA polymorphisms in low penetrance genes have emerged as genetic factors that seem to modulate an individual's susceptibility to malignancy. Genetic studies, which lead to a true association, are expected to increase understanding of the pathogenesis of each malignancy and to be a powerful tool for prevention and prognosis in the future. Here, we review the findings of genetic association studies of gene polymorphisms in gynecologic cancer with special reference to glutathione-S-transferase, FAS/CD95 and p53 genes including our recent research results.
Assuntos
DNA/genética , Genes p53/genética , Predisposição Genética para Doença/genética , Neoplasias dos Genitais Femininos/genética , Glutationa Transferase/genética , Polimorfismo Genético , Receptor fas/genética , Alelos , Códon/genética , Feminino , Genótipo , Humanos , Regiões Promotoras Genéticas/genéticaRESUMO
To investigate the clinical significance of human wings apart-like (hWAPL) genetic polymorphisms in cervical carcinogenesis. hWAPL polymorphisms and human papillomavirus (HPV) types were examined in 175 cervical smears of exfoliated cervical cell samples using a real-time polymerase chain reaction system. A significant difference was detected in the frequency of the CC genotype between the HPV(+) low-grade squamous intraepithelial lesion (LSIL) and high-grade squamous intraepithelial lesion (HSIL) groups [Odds ratio 0.21, 95% confidence interval (CI) 0.0723-0.61; P = 0.0029]. A significant difference was noted in the frequency of the CC genotype between the high-risk HPV-positive LSIL and HSIL groups (odds ratio 0.2955, 95% CI 0.0893-0.9771; P = 0.0414). The CC genotype of hWAPL gene promoter polymorphism may be associated with cervical carcinogenesis.