Assuntos
Antígenos de Neoplasias/genética , Biomarcadores Tumorais/genética , Genes Supressores de Tumor , Transtornos Mieloproliferativos/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Ligação a DNA , Feminino , Genes p53 , Humanos , Janus Quinase 2/genética , Masculino , Pessoa de Meia-Idade , Mutação , Fatores de TranscriçãoRESUMO
MicroRNAs (miRNAs) regulate mRNA stability and protein expression, and certain miRNAs have been demonstrated to act either as oncogenes or tumor suppressors. Differential miRNA expression signatures have been documented in many human cancers but the role of miRNAs in endometrioid endometrial cancer (EEC) remains poorly understood. This study identifies significantly dysregulated miRNAs of EEC cells, and characterizes their impact on the malignant phenotype. We studied the expression of 365 human miRNAs using Taqman low density arrays in EECs and normal endometriums. Candidate differentially expressed miRNAs were validated by quantitative real-time PCR. Expression of highly dysregulated miRNAs was examined in vitro through the effect of anti-/pre-miRNA transfection on the malignant phenotype. We identified 16 significantly dysregulated miRNAs in EEC and 7 of these are novel findings with respect to EEC. Antagonizing the function of miR-7, miR-194 and miR-449b, or overexpressing miR-204, repressed migration, invasion and extracellular matrix-adhesion in HEC1A endometrial cancer cells. FOXC1 was determined as a target gene of miR-204, and two binding sites in the 3'-untranslated region were validated by dual luciferase reporter assay. FOXC1 expression was inversely related to miR-204 expression in EEC. Functional analysis revealed the involvement of FOXC1 in migration and invasion of HEC1A cells. Our results present dysfunctional miRNAs in endometrial cancer and identify a crucial role for miR-204-FOXC1 interaction in endometrial cancer progression. This miRNA signature offers a potential biomarker for predicting EEC outcomes, and targeting of these cancer progression- and metastasis-related miRNAs offers a novel potential therapeutic strategy for the disease.
Assuntos
Fatores de Transcrição Forkhead/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/fisiologia , Invasividade Neoplásica , Regiões 3' não Traduzidas , Adesão Celular , Linhagem Celular Tumoral , Movimento Celular , Neoplasias do Endométrio , Endométrio/metabolismo , Feminino , Perfilação da Expressão Gênica , Humanos , Transfecção , Estudos de Validação como AssuntoRESUMO
While ECT is widely used for the management of severe and refractory depression, its utility in bipolar disorder is not extensively studied. The aim of this study was to examine the reported effectiveness of ECT in patients with unipolar and bipolar depression as reported by psychiatrists, nurses and patients (i.e. using objective and subjective measures). The records of 787 consecutive inpatient admissions to the Geelong Clinic, a private psychiatric centre based outside Melbourne, Victoria were reviewed in this file audit. Routine assessment measures were completed at admission and discharge, and included patient rated measures (Medical Outcomes Short Form SF-14 and Depression Anxiety and Stress Scale, DASS), nurse rated measures, (The Health of the Nation Outcome Scale, HoNOS) and a psychiatrist rated measure, the Clinical Global impression scale (CGI). In contrast to individuals with unipolar depression, where improvement was seen on all measures, in bipolar disorder, while improvement in clinician rated measures was seen (CGI, HoNOS), there was an absence of improvement in subjective measures of mood (DASS, SF14). This study suggests that in bipolar disorder, there is a poorer subjective response to ECT than in unipolar disorder.
Assuntos
Transtorno Bipolar/terapia , Transtorno Depressivo/terapia , Eletroconvulsoterapia/métodos , Nível de Saúde , Escalas de Graduação Psiquiátrica/estatística & dados numéricos , Atitude do Pessoal de Saúde , Atitude Frente a Saúde , Transtorno Bipolar/diagnóstico , Transtorno Depressivo/diagnóstico , Manual Diagnóstico e Estatístico de Transtornos Mentais , Feminino , Hospitalização , Humanos , Masculino , Pessoa de Meia-Idade , Inventário de Personalidade/estatística & dados numéricos , Valor Preditivo dos Testes , Psiquiatria/estatística & dados numéricos , Qualidade de Vida , Inquéritos e Questionários , Resultado do Tratamento , VitóriaRESUMO
The common fragile sites (CFSs) are large regions of profound genomic instability found in all individuals. A number of the CFSs have been found to span genes that extend over large genomic regions (>700 kb). The expression of these genes is frequently abrogated in a number of different cancers and several of them have already been shown to function as tumor suppressor genes, both in vitro and in vivo. We analyzed the expression of 14 large CFS genes in two distinct groups of head and neck cancers using real-time RT-PCR. The first were oral tongue squamous cell carcinomas (SCCs) and the second were base of tongue/tonsillar (oropharyngeal) SCCs. These two groups were previously examined for the presence of human papillomavirus (HPV) and while 46% of the oropharyngeal cancers were positive for HPV16 only one of 52 oral cancers contained HPV16 sequences. We observed a distinct pattern of loss of expression of the large CFS genes in the two groups of head and neck cancers. In addition, there was no correlation between the relative instability in different CFS regions and which genes were inactivated. Thus, this report demonstrates another distinction between these two groups of head and neck cancer. In addition, it suggests that there is selection for loss of expression of specific CFS genes in these cancers.
Assuntos
Carcinoma de Células Escamosas/genética , Sítios Frágeis do Cromossomo , Perfilação da Expressão Gênica , Neoplasias Orofaríngeas/genética , Neoplasias da Língua/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Bases , Primers do DNA , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The common fragile sites are regions of profound genomic instability found in all individuals. The full size of each region of instability ranges from under one megabase (Mb) to greater than 10 Mbs. At least half of the CFS regions have been found to span extremely large genes that spanned from 600 kb to greater than 2.0 Mbs. The large CFS genes are also very interesting from a cancer perspective as several of them, including FHIT and WWOX, have already demonstrated the capacity to function as tumor suppressor genes, both in vitro and in vivo. We estimate that there may be 40-50 large genes localized in CFS regions. The expression of a number of the large CFS genes has been previously shown to be lost in many different cancers and this is frequently associated with a worse clinical outcome for patients. To determine if there was selection for the inactivation of different large CFS genes in different cancers, we examined the expression of 13 of the 20 known large CFS genes: FHIT, WWOX, PARK2, GRID2, NBEA, DLG2, RORA isoforms 1 and 4, DAB1, CNTNAP2, DMD, IL1RAPL1, IMMP2L and LARGE in breast, ovarian, endometrial and brain cancers using real-time RT-PCR analysis. Each cancer had a distinct profile of different large CFS genes that were inactivated. Interestingly, in breast, ovarian and endometrial cancers there were some cancers that had inactivation of expression of none or only one of the tested genes, while in other specimens there was inactivation of multiple tested genes. Brain cancers had inactivation of many of the tested genes, a number of which function in normal neurological development. We find that there is no relationship between the frequency that any specific CFS is expressed and the frequency that the gene from that region is inactivated in different cancers. Instead, it appears that different cancers select for the inactivation of different large CFS genes.
Assuntos
Sítios Frágeis do Cromossomo , Neoplasias/genética , Neoplasias Encefálicas/genética , Neoplasias da Mama/genética , Neoplasias do Endométrio/genética , Feminino , Humanos , Hibridização in Situ Fluorescente , Neoplasias Ovarianas/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Recent advances in the fields of immunology, genetics, molecular biology, bioinformatics, and the Human Genome Project have allowed for the emergence of the field of vaccinomics. Vaccinomics encompasses the fields of immunogenetics and immunogenomics as applied to understanding the mechanisms of heterogeneity in immune responses to vaccines. In this study, we examine the role of HLA genes, cytokine genes, and cell surface receptor genes as examples of how genetic polymorphism leads to individual and population variations in immune responses to vaccines. In turn, this data, in concert with new high-throughput technology, inform the immune-response network theory to vaccine response. Such information can be used in the directed and rational development of new vaccines, and this new golden age of vaccinology has been termed "predictive vaccinology", which will predict the likelihood of a vaccine response or an adverse response to a vaccine, the number of doses needed and even whether a vaccine is likely to be of benefit (i.e., is the individual at risk for the outcome for which the vaccine is being administered?).
Assuntos
Formação de Anticorpos/genética , Citocinas/genética , Antígenos HLA/genética , Imunogenética/métodos , Receptores de Superfície Celular/genética , Vacinas/imunologia , Animais , Biometria/métodos , Biologia Computacional/métodos , Epigênese Genética , Genômica , Haplótipos , Heterozigoto , Humanos , Hibridização de Ácido Nucleico/métodos , Análise de Sequência com Séries de Oligonucleotídeos , Farmacogenética , Polimorfismo Genético , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA/métodos , Estudos em Gêmeos como Assunto , Vacinas Virais/imunologiaRESUMO
Common fragile sites (CFSs) are large regions of profound genomic instability found in all individuals. Spanning the center of the two most frequently expressed CFS regions, FRA3B (3p14.3) and FRA16D (16q23.2), are the 1.5 Mb FHIT gene and the 1.0 Mb WWOX gene. These genes are frequently deleted and/or altered in many different cancers. Both FHIT and WWOX have been demonstrated to function as tumor suppressors, both in vitro and in vivo. A number of other large CFS genes have been identified and are also frequently inactivated in multiple cancers. Based on these data, several additional very large genes were tested to determine if they were derived from within CFS regions, but DCC and RAD51L1 were not. However, the 2.0 Mb DMD gene and its immediately distal neighbor, the 1.8 Mb IL1RAPL1 gene are CFS genes contained within the FRAXC CFS region (Xp21.2-->p21.1). They are abundantly expressed in normal brain but were dramatically underexpressed in every brain tumor cell line and xenograft (derived from an intracranial model of glioblastoma multiforme) examined. We studied the expression of eleven other large CFS genes in the same panel of brain tumor cell lines and xenografts and found reduced expression of multiple large CFS genes in these samples. In this report we show that there is selective loss of specific large CFS genes in different cancers that does not appear to be mediated by the relative instability within different CFS regions. Further, the inactivation of multiple large CFS genes in xenografts and brain tumor cell lines may help to explain why this type of cancer is highly aggressive and associated with a poor clinical outcome.
Assuntos
Neoplasias Encefálicas/genética , Sítios Frágeis do Cromossomo/genética , Distrofina/genética , Regulação Neoplásica da Expressão Gênica , Proteína Acessória do Receptor de Interleucina-1/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Ligação a DNA/genética , Regulação para Baixo , Distrofina/metabolismo , Feminino , Humanos , Hibridização in Situ Fluorescente , Proteína Acessória do Receptor de Interleucina-1/metabolismo , Masculino , Pessoa de Meia-Idade , Transplante Heterólogo , Células Tumorais CultivadasRESUMO
Endometrial cancer is the third most common gynecologic malignancy and the ninth most common malignancy for females overall in Hong Kong. Approximately 80% or more of these cancers are endometrioid endometrial adenocarcinomas. The aim of this study was to reveal genes contributing to the development of endometrioid endometrial cancer, which may impact diagnosis, prognosis and treatment of the disease. Whole-genome gene expression analysis was completed for a set of 55 microdissected sporadic endometrioid endometrial adenocarcinomas and 29 microdissected normal endometrium specimens using the Affymetrix Human U133 Plus 2.0 oligonucleotide microarray. Selected genes of interest were validated by quantitative real-time-polymerase chain reaction (qRT-PCR). Pathway analysis was performed to reveal gene interactions involved in endometrial tumorigenesis. Unsupervised hierarchical clustering displayed a distinct separation between the endometrioid adenocarcinomas and normal endometrium samples. Supervised analysis identified 117 highly differentially regulated genes (>or=4.0-fold change), which distinguished the endometrial cancer specimens from normal endometrium. Twelve novel genes including DKK4, ZIC1, KIF1A, SAA2, LOC16378, ALPP2, CCL20, CXCL5, BST2, OLFM1, KLRC1 and MBC45780 were deregulated in the endometrial cancer, and further validated in an independent set of 56 cancer and 29 normal samples using qRT-PCR. In addition, 10 genes were differentially regulated in late-stage cancer, as compared to early-stage disease, and may be involved in tumor progression. Pathway analysis of the expression data from this tumor revealed an interconnected network consisting of 21 aberrantly regulated genes involved in angiogenesis, cell proliferation and chromosomal instability. The results of this study highlight the molecular features of endometrioid endometrial cancer and provide insight into the events underlying the development and progression of endometrioid endometrial cancer.
Assuntos
Neoplasias do Endométrio/metabolismo , Perfilação da Expressão Gênica , Genoma , Transdução de Sinais , Neoplasias do Endométrio/genética , Feminino , Hong Kong , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
UNLABELLED: The objective of the present preliminary study was to determine if a difference in the pattern of gene expression exists between tumors that were subsequently found to be sensitive to radiotherapy and tumors found to be resistant to radiotherapy. PATIENTS AND METHODS: A total of 16 patients with invasive squamous cell carcinoma of the uterine cervix were included in this study. All patients were treated with standardized radiotherapy alone. Ten of the tumors were clinically radiosensitive and six were radioresistant. Total RNA, extracted from tumor specimens obtained prior to treatment, was hybridized onto an oligonucleotide microarray with probe sets complementary to over 20,000 transcripts. The genes were first subjected to a statistical filter to identify genes with statistically significant differential expression levels between those that were radiosensitive and those that were radioresistant. A back-propagation neural network was then constructed to model the differences so that patterns could be easily identified. RESULTS: Although a number of genes were found to express differentially between radiosensitive and radioresistant tumors; the 10 most discriminating genes were used to construct the model. Using the expressions from these 10 genes, we found that neural networks constructed from random subsets of the whole data were capable of predicting radiotherapy responses in the remaining subset, which appears stable within the dataset. DISCUSSION: This study shows that such an approach has the potential to differentiate tumor radiosensitivity, although confirmation of such a pattern using other larger independent datasets is necessary before firm conclusions can be drawn.
Assuntos
Carcinoma de Células Escamosas/genética , Regulação Neoplásica da Expressão Gênica , Proteínas de Neoplasias/genética , Tolerância a Radiação/genética , Neoplasias do Colo do Útero/genética , Adulto , Idoso , Carcinoma de Células Escamosas/radioterapia , Feminino , Perfilação da Expressão Gênica , Humanos , Pessoa de Meia-Idade , Invasividade Neoplásica/patologia , Análise de Sequência com Séries de Oligonucleotídeos , RNA Neoplásico , Células Tumorais Cultivadas , Neoplasias do Colo do Útero/radioterapiaRESUMO
Common fragile sites (CFSs) are large genomic regions present in all individuals that are highly unstable and prone to breakage and rearrangement, especially in cancer cells with genomic instability. Eight of the 90 known CFSs have been precisely defined and five of these span genes that extend from 700 kb to over 1.5 Mb of genomic sequence. Although these genes reside within some of the most unstable chromosomal regions in the human genome, they are highly conserved evolutionarily. These genes are targets for large chromosomal deletions and rearrangements in cancer and are frequently inactivated in multiple tumor types. There is also an association between these genes and cellular responses to stress. Based upon the association between large genes and CFSs, we began to systematically test other large genes derived from chromosomal regions that were known to contain a CFS. In this study, we demonstrate that the 730 kb retinoic acid receptor-related orphan receptor alpha (RORA) gene is derived from the middle of the FRA15A (15q22.2) CFS. Although this gene is expressed in normal breast, prostate and ovarian epithelium, it is frequently inactivated in cancers that arise from these organs. RORA was previously shown to be involved in the cellular response to hypoxia and here we demonstrate changes in the amount of RORA message produced in cells exposed to a variety of different cellular stresses. Our results demonstrate that RORA is another very large CFS gene that is inactivated in multiple tumors. In addition, RORA appears to play a critical role in responses to cellular stress, lending further support to the idea that the large CFS genes function as part of a highly conserved stress response network that is uniquely susceptible to genomic instability in cancer cells.
Assuntos
Sítios Frágeis do Cromossomo , Cromossomos Humanos Par 15/genética , Genes Neoplásicos/genética , Estresse Oxidativo , Receptores do Ácido Retinoico/fisiologia , Quebra Cromossômica , Deleção Cromossômica , Instabilidade Genômica , Humanos , Dados de Sequência Molecular , Membro 1 do Grupo F da Subfamília 1 de Receptores Nucleares , Receptores Citoplasmáticos e Nucleares , TransativadoresRESUMO
An U-series isotopic study was carried out in the waters of the Avon Valley, Nova Scotia. The fresh and acidic recharge waters flow rapidly through the watershed composed of a granitic highland and a sedimentary, largely carbonate, lowland plain, before draining to the sea. There is no significant anthropogenic pollution; but, naturally elevated U levels can be encountered within the bedrock. Nonetheless, the U concentrations of the surface and groundwater are low (generally within the range of several hundredths to several tenths of a microg l(-1)), except in the proximity to weathering of U mineralization. The dissolved U in the surface waters appears to be stabilized by organic rather than inorganic complexes. Both the groundwaters and surface waters have similar (234)U/(238)U activity ratios that rarely deviate from secular equilibrium by more than 20% throughout the watershed. The magnitude of the (234)U/(238)U activity ratio is not determined by lithology but rather by the weathering mechanism, the high rate of flushing, and the leaching of local U mineralization. Dissolved Ra is consistently absent. The dissolved Rn concentrations, though variable, are measurable even in surface waters. This may be due to a continual degassing from the U-enriched bedrock or release from local sites of U mineralization underlying the surface water sources.
Assuntos
Urânio/análise , Água/química , Monitoramento Ambiental , Fenômenos Geológicos , Geologia , Nova Escócia , Rios , SolubilidadeRESUMO
Chronic infections with the hepatitis B virus (HBV) and high-risk human papillomaviruses (HPVs) are important risk factors for hepatocellular carcinoma (HCC) and cervical cancer (CC), respectively. HBV and HPV are DNA viruses that almost invariably integrate into the host genome in invasive tumors. The viral integration sites occur throughout the genome, leading to the presumption that there are no preferred sites of integration. A number of viral integrations have been shown to occur within the vicinity of important cancer-related genes. In studies of HBV-induced HCC and HPV-induced CC, we have identified two HBV and three HPV integrations into the human telomerase reverse transcriptase (hTERT) gene. Detailed characterization of the integrations revealed that four integrations occurred within the hTERT promoter and upstream region and the fifth integration occurred in intron 3 of the hTERT gene. None of the integrations altered the hTERT coding sequence and all resulted in juxtaposition of viral enhancers near hTERT, with potential activation of hTERT expression. Our work supports the hypothesis that the sites of oncogenic viral integration are nonrandom and that genes at the sites of viral integration may play important roles in carcinogenesis.
Assuntos
Vírus da Hepatite B/genética , Neoplasias Hepáticas/virologia , Papillomaviridae/genética , Telomerase/genética , Neoplasias do Colo do Útero/virologia , Integração Viral , Sequência de Bases , Proteínas de Ligação a DNA , Feminino , Regulação Enzimológica da Expressão Gênica , Humanos , Neoplasias Hepáticas/enzimologia , Neoplasias Hepáticas/etiologia , Dados de Sequência Molecular , Células Tumorais Cultivadas , Neoplasias do Colo do Útero/enzimologia , Neoplasias do Colo do Útero/etiologiaRESUMO
The possibility of horizontal transmission of diflubenzuron and fipronil was assessed in rangeland grasshoppers. Laboratory studies of Melanoplus sanguinipes demonstrated that fipronil was horizontally transferred at lethal levels (p < 0.05) via cannibalism through four passages when the initial dose applied to a food source was 250 times the label rate for rangeland grasshopper and locust control (label rate is 4 g AI/ha). Mortality was 100% on the first three passages through cannibalism. At 25 and 1 times the label rate, fipronil was lethal (p < 0.05) only on the first cannibalistic passage. Diflubenzuron generated significant (p < 0.05) mortality via horizontal transmission through two passages when the initial dose applied to a food source was 2,000 times the label rate for rangeland grasshopper control (label rate is 8.71 g AI/ha). There was 100% mortality in the first passage via cannibalism. At 250 and 25 times the label rate, diflubenzuron was lethal only on the first cannibalistic passage. Field applications of these two acridicides followed by collection of cadavers (Amphitornus coloradus and Ageneotettix deorum) that were fed to M. sanguinipes in the laboratory revealed that fipronil (25 times the label rate) generated significant (p < 0.05) mortality through two passages and diflubenzuron (label rate) caused no mortality via necrophagy. Tenebrionid beetles fed grasshopper cadavers collected from the field application of fipronil yielded 45% mortality, compared with 25% mortality in the controls. These findings suggest that horizontal and trophic transfer probably play a nominal ecotoxicological role in rangeland grasshopper control programs with diflubenzuron, but the transfer of fipronil to grasshoppers, scavengers, and natural enemies via necrophagy may increase both the efficacy of control programs and their environmental affects.
Assuntos
Besouros/fisiologia , Diflubenzuron/toxicidade , Comportamento Alimentar/fisiologia , Gafanhotos/fisiologia , Controle de Insetos/métodos , Pirazóis/toxicidade , Animais , CanibalismoRESUMO
On the afternoon of 7 May 1915 the Cunard Liner Lusitania was torpedoed by a German U-boat whilst en route to Liverpool. Consequently in sank with the loss of 1,195 lives. A series of investigative and formal research exercises, undertaken in the light of eye-witness accounts of the events surrounding that tragedy, has given rise to a number of hypotheses concerning the loss of this vessel. In this paper all the available evidence concerning the attack, and the one or more explosions that occurred, is summarised. The conclusions regarding the torpedo effects and the most likely causes of the secondary events are reported.
RESUMO
The mechanisms by which prostate cancer (PCa) cells progress to a hormone refractory state are poorly understood. The progression process under androgen ablation conditions involves the survival of at least a portion of malignant cells and their eventual proliferation in an androgen-independent manner. The goal of this study was to investigate the role of PI3K signaling in such a progression. Using an in vitro model of androgen ablation, we show that after removal of androgen support, the human PCa cell line LNCaP initially arrested in G(1) and trans-differentiated into neuroendocrine-like cells that eventually resumed androgen-independent proliferation. Both acute and chronic androgen ablation resulted in an increase in basal levels of PI3K and Akt activity, which were sustained throughout the progression process. Under these conditions, inhibition of PI3K, pharmacologically or with ectopic expression of PTEN, arrested cell proliferation and blocked progression to the androgen-independent state. In contrast, LNCaP cells in the presence of androgens were marginally sensitive to PI3K inhibition. During the chronic stage of androgen deprivation, androgen-independent proliferation correlated with diminished p27(kip1) protein levels, whereas PI3K and Akt activity remained elevated. At this stage, PI3K inhibition rapidly triggered accumulation of p27(kip1), cell cycle arrest, and cell death. PI3K modulated p27(kip1) levels at least in part by regulating its rate of degradation. Taken together, these data show that androgen ablation alone can increase PI3K-Akt activation, which supports survival after acute androgen ablation and proliferation during chronic androgen deprivation. Successful progression to the androgen-independent state in the LNCaP cell line model requires intact PI3K signaling.
Assuntos
Androgênios/fisiologia , Fosfatidilinositol 3-Quinases/fisiologia , Neoplasias da Próstata/fisiopatologia , Proteínas Serina-Treonina Quinases , Transdução de Sinais/fisiologia , Antagonistas de Androgênios/farmacologia , Proteínas de Ciclo Celular/metabolismo , Divisão Celular/fisiologia , Sobrevivência Celular/fisiologia , Cromonas/farmacologia , Meios de Cultura/farmacologia , Inibidor de Quinase Dependente de Ciclina p27 , Progressão da Doença , Inibidores Enzimáticos/farmacologia , Humanos , Masculino , Morfolinas/farmacologia , Inibidores de Fosfoinositídeo-3 Quinase , Proteínas Proto-Oncogênicas/fisiologia , Proteínas Proto-Oncogênicas c-akt , Células Tumorais Cultivadas , Proteínas Supressoras de Tumor/metabolismoRESUMO
In the United States, ovarian cancer is the fourth most common cause of cancer-related deaths among women. The most important prognostic factor for this cancer is tumor stage, or extent of disease at diagnosis. Although women with low-stage tumors have a relatively good prognosis, most women diagnosed with late-stage disease eventually succumb to their cancer. In an attempt to understand early events in ovarian carcinogenesis, and to explore steps in its progression, we have applied multiple molecular genetic techniques to the analysis of 21 early-stage (stage I/II) and 17 advanced-stage (stage III/IV) ovarian tumors. These techniques included expression profiling with cDNA microarrays containing approximately 18,000 expressed sequences, and comparative genomic hybridization to address the chromosomal locations of copy number gains as well as losses. Results from the analysis indicate that early-stage ovarian cancers exhibit profound alterations in gene expression, many of which are similar to those identified in late-stage tumors. However, differences observed at the genomic level suggest differences between the early- and late-stage tumors and provide support for a progression model for ovarian cancer development.
Assuntos
Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Progressão da Doença , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Genoma Humano , Humanos , Estadiamento de Neoplasias , Hibridização de Ácido Nucleico , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Tumorais CultivadasRESUMO
Differential display-PCR between ovarian tumor cell lines and short-term cultures of normal ovarian epithelial cell brushings was used to isolate a differentially expressed transcript and its corresponding gene. The gene, which mapped to 13q14.1, has partial homology in the DNAJ domain to a number of proteins with a similar domain and was designated as methylation-controlled J protein (MCJ). MCJ has the highest similarity to a functionally undefined protein from Caenorhabditis elegans. MCJ is expressed as a 1.2-kb transcript in several adult tissues, with testis showing the highest level of expression. Expression of MCJ was absent in three of seven ovarian cancer cell lines. Similarly, expression analysis using semiquantitative reverse transcription-PCR indicated that 12 of 18 primary ovarian tumors examined had either a complete absence or lower levels of expression of this gene. 5-Aza-2'-deoxycytidine treatment of the OV202 cell line induced MCJ expression in a dose-dependent manner, implicating methylation in this induction. Loss of heterozygosity and methylation-specific PCR analysis revealed that the loss of MCJ expression in primary tumors and cell lines was attributable to deletion of one allele and methylation of the other. To assess the potential functional significance of MCJ down-regulation, the sensitivity of parental (MCJ-nonexpressing) and MCJ-transfected OV167 cells to antineoplastic agents was evaluated. MCJ expression was associated with enhanced sensitivity to paclitaxel, topotecan, and cisplatin, suggesting that MCJ loss may play a role in de novo chemoresistance in ovarian carcinoma. These observations raise the possibility that MCJ loss may: (a) have potential prognostic significance in ovarian cancer; and (b) contribute to the malignant phenotype by conferring resistance to the most commonly used chemotherapeutic agents for ovarian cancer.
Assuntos
Azacitidina/análogos & derivados , Resistência a Múltiplos Medicamentos/genética , Proteínas de Choque Térmico/genética , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/genética , Sequência de Aminoácidos , Azacitidina/farmacologia , Sequência de Bases , Mapeamento Cromossômico , Cisplatino/farmacologia , Clonagem Molecular , DNA de Neoplasias/genética , DNA de Neoplasias/isolamento & purificação , Decitabina , Regulação para Baixo , Resistencia a Medicamentos Antineoplásicos , Feminino , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Proteínas de Choque Térmico HSP40 , Proteínas de Choque Térmico/biossíntese , Humanos , Hibridização in Situ Fluorescente , Perda de Heterozigosidade , Dados de Sequência Molecular , Neoplasias Ovarianas/metabolismo , Oximas , Paclitaxel/farmacologia , Piperazinas , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Topotecan/farmacologia , Transfecção , Células Tumorais CultivadasRESUMO
Cytogenetic, molecular and functional analysis has shown that chromosome region 6q27 harbors a senescence inducing gene and a tumor suppressor gene involved in several solid and hematologic malignancies. We have cloned at 6q27 and characterized the RNASE6PL gene which belongs to a family of cytoplasmic RNases highly conserved from plants, to man. Analysis of 55 primary ovarian tumors and several ovarian tumor cell lines indicated that the RNASE6PL gene is not mutated in tumor tissues, but its expression is significantly reduced in 30% of primary ovarian tumors and in 75% of ovarian tumor cell lines. The promoter region of the gene was unaffected in tumors cell lines. Transfection of RNASE6PL cDNA into HEY4 and SG10G ovarian tumor cell lines suppressed tumorigenicity in nude mice. When tumors were induced by RNASE6PL-transfected cells, they completely lacked expression of RNASE6PL cDNA. Tumorigenicity was suppressed also in RNASE6PL-transfected pRPcT1/H6cl2T cells, derived from a human/mouse monochromosomic hybrid carrying a human chromosome 6 deleted at 6q27. Moreover, 63.6% of HEY4 clones and 42.8% of the clones of XP12ROSV, a Xeroderma pigmentosum SV40-immortalized cell line, transfected with RNASE6PL cDNA, developed a marked senescence process during in vitro growth. We therefore propose that RNASE6PL may be a candidate for the 6q27 senescence inducing and class II tumor suppressor gene in ovarian cancer.
Assuntos
Carcinoma/genética , Cromossomos Humanos Par 6/genética , Genes Supressores de Tumor , Neoplasias Ovarianas/genética , Ribonucleases/genética , Proteínas Supressoras de Tumor , Animais , Senescência Celular/genética , Clonagem Molecular , Ilhas de CpG , Metilação de DNA , Feminino , Humanos , Células Híbridas , Camundongos , Camundongos Nus , Dados de Sequência Molecular , Proteínas de Neoplasias/genética , Estadiamento de Neoplasias , RNA de Transferência de Serina , Distribuição TecidualRESUMO
Sodium butyrate (NaBu) was shown to induce differentiation and apoptosis in the human pancreatic cancer cell line AsPC-1. A suppression subtractive hybridization-based technique was used to identify genes induced by NaBu. A novel cDNA was found to be highly up-regulated in AsPC-1 cells in response to NaBu. The gene expresses a 1.65-kb mRNA encoding a protein with an open reading frame of 422 amino acids. It has an intermediate filament signature sequence and extensive homology to type I keratins. Sequence comparison with known keratins indicated that the gene shares 42-46% amino acid identity and 60-65% similarity within the alpha-helical rod domain. The gene is named K23 (for human type I Keratin 23, KRT23). K23 mRNA was highly induced by NaBu in different pancreatic cancer cells. Trichostatin A (TSA), a potent and specific inhibitor of histone deacetylase, similarly induced K23 mRNA expression. Treatment with either actinomycin D or cycloheximide efficiently blocked the induction of K23 mRNA by NaBu/TSA. These results indicate that K23 mRNA induction by NaBu or TSA is a downstream event of histone hyperacetylation. We also demonstrated that expression of p21(WAF1/CIP1) antisense RNA efficiently blocked the induction of K23 mRNA induced by NaBu. Our results suggest that K23 is a novel member of the acidic keratin family that is induced in pancreatic cancer cells undergoing differentiation by a mechanism involving histone hyperacetylation. p21(WAF1/CIP1) serves as an important mediator during the induction process of K23 by NaBu.
Assuntos
Inibidores Enzimáticos/farmacologia , Inibidores de Histona Desacetilases , Queratinas/biossíntese , Neoplasias Pancreáticas/enzimologia , Neoplasias Pancreáticas/patologia , Acetilação , Sequência de Aminoácidos , Apoptose/efeitos dos fármacos , Apoptose/genética , Sequência de Bases , Butiratos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Clonagem Molecular , Inibidor de Quinase Dependente de Ciclina p21 , Ciclinas/biossíntese , Ciclinas/fisiologia , Histona Desacetilases/metabolismo , Histonas/metabolismo , Humanos , Proteínas de Filamentos Intermediários/química , Proteínas de Filamentos Intermediários/genética , Proteínas de Filamentos Intermediários/isolamento & purificação , Queratinas/química , Queratinas/genética , Queratinas Tipo I , Dados de Sequência Molecular , Neoplasias Pancreáticas/metabolismo , RNA Mensageiro/biossíntese , Homologia de Sequência de Aminoácidos , Células Tumorais CultivadasRESUMO
OBJECTIVE: To investigate the extent of knowledge of Australian general practitioners (GPs) in terms of critical information about adolescent suicide. METHOD: GPs knowledge about adolescent suicide was assessed using the 39-item Adolescent Suicide Behaviour Questionnaire (ASBQ), distributed to all GPs listed on the Medical Practitioners Board register of the Australian state of Victoria. Forty-three percent of GPs (n = 1694) completed and returned the survey. RESULTS: GPs scored, on average, 71 percent of the questionnaire items correct. The mean level of incorrect and uncertain responses was 14 percent and 15 percent, respectively. However, there were wide differences in the number of correct scores between individual GPs, with some respondents scoring as few as four items correct and others as many as 38 items correct. Analysis of knowledge rates within ABSQ content domains showed that GPs were generally well informed about adolescent suicide in relation to precipitating factors, and less well informed about the remaining content domains. CONCLUSIONS: Australian GPs are, in general, moderately well informed, and are in a unique position to identify those at risk and to provide appropriate intervention (or referral). However, there is considerable variability in the accuracy of beliefs about adolescent suicide, with some GPs demonstrating excellent knowledge levels but others holding little accurate information. The extent of this variability in knowledge is a cause for concern. Findings highlight the need for ongoing education of GPs as an essential component of prevention strategies for youth suicide.