RESUMO
BACKGROUND: Androgen receptor (AR)-gene amplification, found in 20-30% of castration-resistant prostate cancer (CRPCa) is proposed to develop as a consequence of hormone-deprivation therapy and be a prime cause of treatment failure. Here we investigate AR-gene amplification in cancers before hormone deprivation therapy. METHODS: A tissue microarray (TMA) series of 596 hormone-naive prostate cancers (HNPCas) was screened for chromosome X and AR-gene locus-specific copy number alterations using four-colour fluorescence in situ hybridisation. RESULTS: Both high level gain in chromosome X (≥4 fold; n=4, 0.7%) and locus-specific amplification of the AR-gene (n=6, 1%) were detected at low frequencies in HNPCa TMAs. Fluorescence in situ hybridisation mapping whole sections taken from the original HNPCa specimen blocks demonstrated that AR-gene amplifications exist in small foci of cells (≤ 600 nm, ≤1% of tumour volume). Patients with AR gene-locus-specific copy number gains had poorer prostate cancer-specific survival. CONCLUSION: Small clonal foci of cancer containing high level gain of the androgen receptor (AR)-gene develop before hormone deprivation therapy. Their small size makes detection by TMA inefficient and suggests a higher prevalence than that reported herein. It is hypothesised that a large proportion of AR-amplified CRPCa could pre-date hormone deprivation therapy and that these patients would potentially benefit from early total androgen ablation.
Assuntos
Hibridização in Situ Fluorescente/métodos , Neoplasias de Próstata Resistentes à Castração/genética , Receptores Androgênicos/genética , Idoso , Amplificação de Genes , Humanos , Estimativa de Kaplan-Meier , Masculino , Prognóstico , Neoplasias de Próstata Resistentes à Castração/patologia , Receptores Androgênicos/metabolismo , Análise de SobrevidaRESUMO
BACKGROUND: The natural history of prostate cancer is highly variable and difficult to predict. We report on the prognostic value of phosphatase and tensin homologue (PTEN) loss in a cohort of 675 men with conservatively managed prostate cancer diagnosed by transurethral resection of the prostate. METHODS: The PTEN status was assayed by immunohistochemistry (PTEN IHC) and fluorescent in situ hybridisation (PTEN FISH). The primary end point was death from prostate cancer. RESULTS: The PTEN IHC loss was observed in 18% cases. This was significantly associated with prostate cancer death in univariate analysis (hazard ratio (HR)=3.51; 95% CI 2.60-4.73; P=3.1 × 10(-14)). It was highly predictive of prostate cancer death in the 50% of patients with a low risk score based on Gleason score, PSA, Ki-67 and extent of disease (HR=7.4; 95% CI 2.2-24.6; P=0.012) ), but had no prognostic value in the higher risk patients. The PTEN FISH loss was only weakly associated with PTEN IHC loss (κ=0.5). Both PTEN FISH loss and amplification were univariately predictive of death from prostate cancer, but this was not maintained in the multivariate analyses. CONCLUSION: In low-risk patients, PTEN IHC loss adds prognostic value to Gleason score, PSA, Ki-67 and extent of disease.
Assuntos
Adenocarcinoma/diagnóstico , Biomarcadores Tumorais/genética , PTEN Fosfo-Hidrolase/genética , Neoplasias da Próstata/diagnóstico , Adenocarcinoma/genética , Adenocarcinoma/mortalidade , Adenocarcinoma/cirurgia , Adulto , Idoso , Biomarcadores Tumorais/metabolismo , Regulação Neoplásica da Expressão Gênica , Inativação Gênica/fisiologia , Humanos , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Antígeno Ki-67/metabolismo , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , PTEN Fosfo-Hidrolase/metabolismo , Valor Preditivo dos Testes , Prognóstico , Antígeno Prostático Específico/metabolismo , Neoplasias da Próstata/genética , Neoplasias da Próstata/mortalidade , Neoplasias da Próstata/cirurgia , Ressecção Transuretral da PróstataRESUMO
Human synovial sarcomas contain a recurrent and specific chromosomal translocation t(X;18)(p11.2;q11.2). By screening a synovial sarcoma cDNA library with a yeast artificial chromosome spanning the X chromosome breakpoint, we have identified a hybrid transcript that contains 5' sequences (designated SYT) mapping to chromosome 18 and 3' sequences (designated SSX) mapping to chromosome X. An SYT probe detected genomic rearrangements in 10/13 synovial sarcomas. Sequencing of cDNA clones shows that the normal SYT gene encodes a protein rich in glutamine, proline and glycine, and indicates that in synovial sarcoma rearrangement of the SYT gene results in the formation of an SYT-SSX fusion protein. Both SYT and SSX failed to exhibit significant homology to known gene sequences.
Assuntos
Cromossomos Humanos Par 18 , Sarcoma Sinovial/genética , Translocação Genética , Cromossomo X , Sequência de Aminoácidos , Sequência de Bases , Primers do DNA/genética , DNA Complementar/genética , DNA de Neoplasias/genética , Rearranjo Gênico , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , RNA Neoplásico/genética , Mapeamento por Restrição , Células Tumorais CultivadasRESUMO
The allele sizes of polymorphic microsatellite repeats in DNA from human cancers were compared to normal DNA from the same patients. In 16 out of 196 paired samples (8%), we found evidence of an extra allele of a different size in the tumour which was not present in the normal DNA. Sequence analysis confirmed that the extra allele originates from the appropriate locus and that the size change is attributable to alteration in the number of repeat units. This form of instability was more common in tri- and tetranucleotide repeats than in dinucleotide repeats. In any single tumour sample only one repeat in the set examined was abnormal, the remainder showing identical patterns in normal and tumour DNA or evidence of allele loss. The pattern of instability in diverse types of cancer differs from that reported in colorectal neoplasms.
Assuntos
DNA Satélite/genética , Neoplasias/genética , Polimorfismo Genético , Sequências Repetitivas de Ácido Nucleico/genética , Alelos , Sequência de Bases , Neoplasias Encefálicas/genética , Neoplasias da Mama/genética , DNA de Neoplasias/genética , Feminino , Humanos , Masculino , Dados de Sequência Molecular , Distrofia Miotônica/genética , Oligodesoxirribonucleotídeos , Neoplasias Ovarianas/genética , Sarcoma/genética , Análise de Sequência de DNARESUMO
Testicular germ-cell tumours (TGCT) affect 1 in 500 men and are the most common cancer in males aged 15-40 in Western European populations. The incidence of TGCT has risen dramatically over the last century. Known risk factors for TGCT include a history of undescended testis (UDT), testicular dysgenesis, infertility, previously diagnosed TGCT (ref. 7) and a family history of the disease. Brothers of men with TGCT have an 8-10-fold risk of developing TGCT (refs 8,9), whereas the relative risk to fathers and sons is fourfold (ref. 9). This familial relative risk is much higher than that for most other types of cancer. We have collected samples from 134 families with two or more cases of TGCT, 87 of which are affected sibpairs. A genome-wide linkage search yielded a heterogeneity lod (hlod) score of 2.01 on chromosome Xq27 using all families compatible with X inheritance. We obtained a hlod score of 4.7 from families with at least one bilateral case, corresponding to a genome-wide significance level of P=0.034. The proportion of families with UDT linked to this locus was 73% compared with 26% of families without UDT (P=0.03). Our results provide evidence for a TGCT susceptibility gene on chromosome Xq27 that may also predispose to UDT.
Assuntos
Predisposição Genética para Doença/genética , Germinoma/genética , Neoplasias Testiculares/genética , Cromossomo X , Adolescente , Adulto , Mapeamento Cromossômico , Família , Feminino , Marcadores Genéticos , Germinoma/epidemiologia , Humanos , Incidência , Escore Lod , Masculino , Fatores de Risco , Neoplasias Testiculares/epidemiologiaRESUMO
BACKGROUND: Previously, using gene-knockdown techniques together with genome expression array analysis, we showed the gene protein Kinase C (PKC)-zeta (PRKCZ) to mediate the malignant phenotype of human prostate cancer. However, according to NCBI, the gene has undergone several major iterations. Therefore, to understand the relationship between its structure and biological activities, we have analysed its expressed sequence in prostate cancer cell lines and tissues. METHODS: Transcriptome-walking and targeted PCR were used to sequence the mRNA transcribed from PRKCZ. Hydropathy analysis was employed to analyse the hypothetical protein sequence subsequently translated and to identify an appropriate epitope to generate a specific monoclonal antibody. RESULTS: A novel sequence was identified within the 3'-terminal domain of human PRKCZ that, in prostate cancer cell lines and tissues, is expressed during transcription and thereafter translated into protein (designated PKC-ζ(-PrC)) independent of conventional PKC-ζ(-a). The monoclonal antibody detected expression of this 96 kD protein only within malignant prostatic epithelium. INTERPRETATION: Transcription and translation of this gene sequence, including previous intronic sequences, generates a novel specific biomarker of human prostate cancer. The presence of catalytic domains characteristic of classic PKC-ß and atypical PKC-ι within PKC-ζ(-PrC) provides a potential mechanism for this PRKCZ variant to modulate the malignant prostatic phenotype out-with normal cell-regulatory control.
Assuntos
Biomarcadores Tumorais/biossíntese , Biomarcadores Tumorais/genética , Neoplasias da Próstata/enzimologia , Neoplasias da Próstata/genética , Proteína Quinase C/biossíntese , Proteína Quinase C/genética , Sequência de Aminoácidos , Sequência de Bases , Biomarcadores Tumorais/metabolismo , Domínio Catalítico , Linhagem Celular , Linhagem Celular Tumoral , Células Epiteliais/metabolismo , Variação Genética , Humanos , Masculino , Dados de Sequência Molecular , Fenótipo , Neoplasias da Próstata/metabolismo , Proteína Quinase C/metabolismo , Splicing de RNA , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Transcrição Gênica , Transcriptoma/genéticaRESUMO
BACKGROUND: The discovery of ERG/ETV1 gene rearrangements and PTEN gene loss warrants investigation in a mechanism-based prognostic classification of prostate cancer (PCa). The study objective was to evaluate the potential clinical significance and natural history of different disease categories by combining ERG/ETV1 gene rearrangements and PTEN gene loss status. METHODS: We utilised fluorescence in situ hybridisation (FISH) assays to detect PTEN gene loss and ERG/ETV1 gene rearrangements in 308 conservatively managed PCa patients with survival outcome data. RESULTS: ERG/ETV1 gene rearrangements alone and PTEN gene loss alone both failed to show a link to survival in multivariate analyses. However, there was a strong interaction between ERG/ETV1 gene rearrangements and PTEN gene loss (P<0.001). The largest subgroup of patients (54%), lacking both PTEN gene loss and ERG/ETV1 gene rearrangements comprised a 'good prognosis' population exhibiting favourable cancer-specific survival (85.5% alive at 11 years). The presence of PTEN gene loss in the absence of ERG/ETV1 gene rearrangements identified a patient population (6%) with poorer cancer-specific survival that was highly significant (HR=4.87, P<0.001 in multivariate analysis, 13.7% survival at 11 years) when compared with the 'good prognosis' group. ERG/ETV1 gene rearrangements and PTEN gene loss status should now prospectively be incorporated into a predictive model to establish whether predictive performance is improved. CONCLUSIONS: Our data suggest that FISH studies of PTEN gene loss and ERG/ETV1 gene rearrangements could be pursued for patient stratification, selection and hypothesis-generating subgroup analyses in future PCa clinical trials and potentially in patient management.
Assuntos
Carcinoma/mortalidade , Proteínas de Ligação a DNA/genética , PTEN Fosfo-Hidrolase/genética , Neoplasias da Próstata/mortalidade , Transativadores/genética , Fatores de Transcrição/genética , Idoso , Carcinoma/diagnóstico , Carcinoma/genética , Carcinoma/metabolismo , Causas de Morte , Estudos de Coortes , Análise Mutacional de DNA/métodos , Proteínas de Ligação a DNA/metabolismo , Loci Gênicos , Humanos , Hibridização in Situ Fluorescente , Masculino , Pessoa de Meia-Idade , Modelos Biológicos , PTEN Fosfo-Hidrolase/metabolismo , Prognóstico , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/genética , Neoplasias da Próstata/metabolismo , Estudos Retrospectivos , Fatores de Risco , Análise de Sobrevida , Análise Serial de Tecidos , Transativadores/metabolismo , Fatores de Transcrição/metabolismo , Regulador Transcricional ERGRESUMO
Substantial evidence now supports the view that epigenetic changes have a role in the development of human prostate cancer. Analyses of the patterns of epigenetic alteration are providing important insights into the origin of this disease and have identified specific alterations that may serve as useful diagnostic and prognostic biomarkers. Examination of cancer methylation patterns supports a stem cell origin of prostate cancer. It is well established that methylation of GSTpi is a marker of prostate cancer, and global patterns of histone marking appear to be linked to cancer prognosis with levels of acetylated histones H3K9, H3K18, and H4K12, and of dimethylated H4R3 and H3K4, dividing low-grade prostate cancer (Gleason 6 or less) into two prognostically separate groups. Elevated levels of several components of the polycomb group protein complex, EZH2, BMI1, and RING1, can also act as biomarkers of poor clinical outcome. Many components of the epigenetic machinery, including histone deacetylase (whose expression level is linked to the TMPRSS2:ERG translocation) and the histone methylase EZH2, are potential therapeutic targets. The recent discovery of the role of small RNAs in governing the epigenetic status of individual genes offers exciting new possibilities in therapeutics and chemoprevention.
Assuntos
Epigênese Genética , Neoplasias da Próstata/genética , Humanos , Masculino , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/terapiaRESUMO
BACKGROUND: This study was performed to test the hypothesis that expression of small heat shock protein Hsp-27 is, at diagnosis, a reliable predictive biomarker of clinically aggressive prostate cancer. METHODS: A panel of tissue microarrays constructed from a well-characterised cohort of 553 men with conservatively managed prostate cancer was stained immunohistochemically to detect Hsp-27 protein. Hsp-27 expression was compared with a series of pathological and clinical parameters, including outcome. RESULTS: Hsp-27 staining was indicative of higher Gleason score (P<0.001). In tissue cores having a Gleason score >7, the presence of Hsp-27 retained its power to independently predict poor clinical outcome (P<0.002). Higher levels of Hsp-27 staining were almost entirely restricted to cancers lacking ERG rearrangements (chi2 trend=31.4, P<0.001), although this distribution did not have prognostic significance. INTERPRETATION: This study has confirmed that, in prostate cancers managed conservatively over a period of more than 15 years, expression of Hsp-27 is an accurate and independent predictive biomarker of aggressive disease with poor clinical outcome (P<0.001). These findings suggest that apoptotic and cell-migration pathways modulated by Hsp-27 may contain targets susceptible to the development of biologically appropriate chemotherapeutic agents that are likely to prove effective in treating aggressive prostate cancers.
Assuntos
Rearranjo Gênico , Proteínas de Choque Térmico HSP27/análise , Neoplasias da Próstata/química , Proteínas Proto-Oncogênicas c-ets/genética , Idoso , Proteínas de Choque Térmico HSP27/fisiologia , Proteínas de Choque Térmico , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Chaperonas Moleculares , Prognóstico , Neoplasias da Próstata/patologiaRESUMO
From a panel of monoclonal antibodies raised against fractions of rat liver nuclear envelopes (NEs), we have identified an antibody, RL30, which reacts with novel nuclear pore complex (NPC) antigens that are not O-glycosylated. By immunofluorescence staining of cultured cells, RL30 reacts exclusively with the NE in a punctate pattern that largely coincides with that of identified NPC proteins. RL30 labels only the cytoplasmic surface of the NPC in immunogold electron microscopy, predominantly in peripheral regions nearby the cytoplasmic ring. In immunoblots of isolated rat liver NEs and cultured rat cells, RL30 recognizes a 265-kD band, as well as a series of 175-265-kD bands in rat liver NEs that are likely to be proteolytic products of p265. Sequencing of peptides from the 175- and 265-kD RL30 antigens of rat liver revealed that they are both closely related to human Tpr, a protein whose amino-terminal 150-250 amino acids appear in oncogenic fusions with the kinase domains of the met, trk, and raf protooncogenes. We found that in vitro translation of human Tpr mRNA yields a major 265-kD band. Considered together, these data indicate that the 265-kD RL30 antigen in the NPC is the rat homologue of Tpr. Interestingly, Tpr contains an exceptionally long predicted coiled coil domain (approximately 1600 amino acids). The localization and predicted structure of Tpr suggest that it is a component of the cytoplasmic fibrils of the NPC implicated in nuclear protein import. Immunofluorescence microscopy shows that during NPC reassembly at the end of mitosis, Tpr becomes concentrated at the NE significantly later than O-linked glycoproteins, including p62. This indicates that reassembly of the NPC after mitosis is a stepwise process, and that the Tpr-containing peripheral structures are assembled later than p62.
Assuntos
Compartimento Celular , Polaridade Celular , Membrana Nuclear/química , Proteínas Proto-Oncogênicas/isolamento & purificação , Sequência de Aminoácidos , Animais , Artefatos , Sequência de Bases , Ativação Enzimática , Imunofluorescência , Humanos , Fígado , Glicoproteínas de Membrana/metabolismo , Microscopia Imunoeletrônica , Mitose/fisiologia , Dados de Sequência Molecular , Complexo de Proteínas Formadoras de Poros Nucleares , Proteínas Quinases/metabolismo , Estrutura Terciária de Proteína , Ratos , Homologia de Sequência de Aminoácidos , Relação Estrutura-AtividadeRESUMO
Early passages of the human teratocarcinoma cell line PA1 are not tumorigenic in nude mice, while late passages are. A transforming gene present in late passages of PA1 cells was isolated as a biologically active molecular clone and is a new isolate of the human rasN locus. Its transforming activity is due to a single G---A (G, guanine; A, adenine) point mutation at the codon for amino acid 12 which changes the codon for glycine so that an aspartic acid residue is expressed. In contrast to late passage PA1 cells (passages 106, 330, and 338), DNA from the PA1 cell line at early passages (passage 36) does not yield rasN foci in DNA transfection assays. Thus, the presence of an activated rasN in PA1 cells correlates with enhanced tumorigenicity of the cell line and, more importantly, may have arisen during cell culture in vitro.
Assuntos
Oncogenes , Teratoma/genética , Animais , Sequência de Bases , Linhagem Celular , Transformação Celular Neoplásica/metabolismo , DNA de Neoplasias/genética , Humanos , Camundongos , Camundongos Nus , Fatores de TempoRESUMO
Tumor induction in athymic nude mice can be used to detect dominant transforming genes in cellular DNA. Mouse NIH 3T3 cells freshly transfected with either cloned Moloney sarcoma proviral DNA or cellular DNA's derived from virally transformed cells induced tumors when injected into athymic nu/nu mice. Tumors were also induced by cells transfected with DNA from two tumor-derived and one chemically transformed human cell lines. The mouse tumors induced by human cell line DNA's contained human DNA sequences, and DNA derived from these tumors was capable of inducing both tumors and foci on subsequent transfection. Tumor induction in nude mice represents a useful new method for the detection and selection of cells transformed by cellular oncogenes.
Assuntos
Transformação Celular Viral , DNA de Neoplasias/genética , Camundongos Nus/fisiologia , Neoplasias Experimentais/genética , Oncogenes , Animais , DNA Viral/genética , Camundongos , Vírus da Leucemia Murina de Moloney/genética , Vírus do Sarcoma Murino/genéticaRESUMO
The mutagenicity of r-8,t-9-dihydroxy-t-10, 11-oxy-8,9,10,11-tetrahydrobenz[a]anthracene (BA-8,9-diol 10, 11-oxide) toward Salmonella typhimurium TA 100 is not decreased by the presence of large amounts of highly purified microsomal or cytosolic epoxide hydrolase. However, highly purified dihydrodiol dehydrogenase inactivates this diol epoxide, which is a major DNA-binding metabolite of benz[a]anthracene. The K-region epoxide, benz[a]anthracene 5,6-oxide (BA 5,6-oxide) is efficiently inactivated by microsomal epoxide hydrolase, is much less readily inactivated by cytosolic epoxide hydrolase, and is not inactivated by dihydrodiol dehydrogenase. This inactivation of a diol epoxide by dihydrodiol dehydrogenase points to a new significance of this enzyme and a new level of control for diol epoxides.
Assuntos
Oxirredutases do Álcool/metabolismo , Benzo(a)Antracenos/metabolismo , Inativação Metabólica , Oxirredutases atuantes sobre Doadores de Grupo CH-CH , Oxirredutases , Benzo(a)Antracenos/farmacologia , Epóxido Hidrolases/metabolismo , Testes de Mutagenicidade , Mutação/efeitos dos fármacos , Salmonella typhimurium/efeitos dos fármacosRESUMO
BACKGROUND: The association between obesity and urinary dysfunction in childhood has been described, albeit through retrospective analysis, making temporal relationships difficult to establish. OBJECTIVE: The objective of this study was to determine risk factors for significant weight gain in children at risk for recurrent urinary tract infections. STUDY DESIGN: A secondary analysis of the Randomized Intervention for Children with Vesicoureteral Reflux and Careful Urinary Tract Infection Evaluation trials was conducted. The outcome of interest in these children was significant increase in body mass index (BMI) percentile (>85th BMI percentile for sex and age) in previously normal-weight children. Multivariable logistic regression was used to determine the independent effects of predetermined risk factors. RESULTS: In total, 446 patients were included in the study. Most patients aged less than 1 year at study entry (229, 51%), and 399 (89%) of patients were female. Eighty-four patients (17%) became clinically overweight. Patients assigned to prophylactic antibiotics were not more likely to gain significant BMI percentiles (adjusted odds ratio [aOR] = 1.1, 95% confidence interval [CI]=0.6-1.8). Significant BMI percentiles were gained in Hispanic/Latino patients compared with whites (aOR = 3.3, 95% CI=1.7-6.4), in children who were infants at study enrollment compared with non-infants (aOR = 2.1, 95% CI=1.2-3.8), and in those with persistent reflux during the study period (aOR = 2.1, 95% CI=1.0-4.3). Neither patients assigned to prophylactic antibiotics (aOR = 1.1, 95% CI=0.6-1.8) nor patients with bladder and bowel dysfunction (BBD) (aOR = 1.2, 95% CI=0.6-2.3) were more likely to gain significant BMI percentiles. DISCUSSION: Significant BMI percentile gain is common in patients at risk for UTIs. Hispanic/Latino ethnicity, persistent reflux, and younger age, specifically infants than non-infants, were identified as independent risk factors for becoming overweight in this population. Exposure to prophylactic antibiotics and BBD were not associated with becoming overweight. CONCLUSION: Risk for becoming overweight should be discussed when managing patients at risk for UTIs, especially in the subpopulations identified.
Assuntos
Obesidade Infantil/epidemiologia , Obesidade Infantil/etiologia , Infecções Urinárias/complicações , Criança , Pré-Escolar , Feminino , Previsões , Humanos , Lactente , Masculino , Estudos Prospectivos , Recidiva , Fatores de RiscoRESUMO
INTRODUCTION: Pediatric bladder and bowel dysfunction (BBD) is a common problem in children. However, the current ability to diagnosis and quantify pediatric BBD is limited as only a few validated instruments exist. In addition, the current questionnaires are limited by their lack of psychometric processing and methods of validation. To address these issues, the authors developed a new questionnaire to objectively diagnose pediatric BBD symptoms. This study aimed to evaluate the performance of this newly devised objective instrument in diagnosing and quantifying the symptomatology of BBD in children. MATERIALS AND METHODS: An 18-item, 5-point questionnaire was developed using both a literature review and expert opinions. The total questionnaire score could range from 0 to 72. Questions were subgrouped into six symptom categories: (1) nocturnal enuresis, (2) lower urinary tract symptoms, (3) urinary holding, (4) infrequent urination, (5) bowel symptoms, and (6) daytime urinary incontinence. The questionnaire also assessed the degree of bother associated with the symptoms. Patients were divided into cases and controls, and these two groups were compared. DISCUSSION/RESULTS: A total of 1265 new patients (758 cases and 507 controls) completed the new BBD questionnaire. The mean age of the whole study cohort was 9.5 years (range, 3-19 years). The total mean questionnaire score was significantly higher at 23 (3-58) in the cases, compared with 8 (0-35) in the controls (p < 0.001) (Summary Figure). Reliability analysis of the 18-item instrument showed a Cronbach's alpha reliability coefficient of 0.80 for the scale. CONCLUSIONS: This new instrument provides a valid and reliable method for diagnosis of pediatric BBD and classification of patients into subcategories of BBD based on their specific symptoms.
Assuntos
Constipação Intestinal/diagnóstico , Enurese/diagnóstico , Programas de Rastreamento/métodos , Psicometria/métodos , Inquéritos e Questionários , Incontinência Urinária/diagnóstico , Adolescente , Criança , Pré-Escolar , Constipação Intestinal/epidemiologia , Enurese/epidemiologia , Feminino , Seguimentos , Humanos , Masculino , Morbidade/tendências , Reprodutibilidade dos Testes , Estudos Retrospectivos , Estados Unidos/epidemiologia , Incontinência Urinária/epidemiologia , Adulto JovemRESUMO
Amplification and overexpression of the E2F3 gene at 6p22 in human bladder cancer is associated with increased tumour stage, grade and proliferation index, and in prostate cancer E2F3 overexpression is linked to tumour aggressiveness. We first used small interfering RNA technology to confirm the potential importance of E2F3 overexpression in bladder cancer development. Knockdown of E2F3 expression in bladder cells containing the 6p22 amplicon strongly reduced the extent of bromodeoxyuridine (BrdU) incorporation and the rate of cellular proliferation. In contrast, knockdown of CDKAL1/FLJ20342, another proposed oncogene, from this amplicon had no effect. Expression cDNA microarray analysis on bladder cancer cells following E2F3 knockdown was then used to identify genes regulated by E2F3, leading to the identification of known E2F3 targets such as Cyclin A and CDC2 and novel targets including pituitary tumour transforming gene 1, Polo-like kinase 1 (PLK1) and Caveolin-2. For both bladder and prostate cancer, we have proposed that E2F3 protein overexpression may cooperate with removal of the E2F inhibitor retinoblastoma tumor suppressor protein (pRB) to drive cellular proliferation. In support of this model, we found that ectopic expression of E2F3a enhanced the BrdU incorporation, a marker of cellular proliferation rate, of prostate cancer DU145 cells, which lack pRB, but had no effect on the proliferation rate of PC3 prostate cancer cells that express wild-type pRB. BrdU incorporation in PC3 cells could, however, be increased by overexpressing E2F3a in cells depleted of pRB. When taken together, these observations indicate that E2F3 levels have a critical role in modifying cellular proliferation rate in human bladder and prostate cancer.
Assuntos
Proliferação de Células , Fator de Transcrição E2F3/fisiologia , Neoplasias da Próstata/patologia , Neoplasias da Bexiga Urinária/patologia , Linhagem Celular Tumoral , Fator de Transcrição E2F3/antagonistas & inibidores , Fator de Transcrição E2F3/biossíntese , Fator de Transcrição E2F3/genética , Humanos , Masculino , Neoplasias da Próstata/genética , Neoplasias da Próstata/metabolismo , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/metabolismoRESUMO
TMPRSS2-ERG gene fusions have recently been reported to be present in a high proportion of human prostate cancers. In the current study, we show that great diversity exists in the precise structure of TMPRSS2-ERG hybrid transcripts found in human prostates. Fourteen distinct hybrid transcripts are characterized, each containing different combinations of sequences from the TMPRSS2 and ERG genes. The transcripts include two that are predicted to encode a normal full-length ERG protein, six that encode N-terminal truncated ERG proteins and one that encodes a TMPRSS2-ERG fusion protein. Interestingly, distinct patterns of hybrid transcripts were found in samples taken from separate regions of individual cancer-containing prostates, suggesting that TMPRSS2-ERG gene fusions may be arising independently in different regions of a single prostate.
Assuntos
Regulação Neoplásica da Expressão Gênica , Variação Genética , Proteínas de Fusão Oncogênica/genética , Próstata/patologia , Neoplasias da Próstata/genética , RNA Mensageiro/genética , Humanos , Masculino , Transdução de SinaisRESUMO
A fluorescence in situ hybridisation (FISH) assay has been used to screen for ETV1 gene rearrangements in a cohort of 429 prostate cancers from patients who had been diagnosed by trans-urethral resection of the prostate. The presence of ETV1 gene alterations (found in 23 cases, 5.4%) was correlated with higher Gleason Score (P=0.001), PSA level at diagnosis (P=<0.0001) and clinical stage (P=0.017) but was not linked to poorer survival. We found that the six previously characterised translocation partners of ETV1 only accounted for 34% of ETV1 re-arrangements (eight out of 23) in this series, with fusion to the androgen-repressed gene C15orf21 representing the commonest event (four out of 23). In 5'-RACE experiments on RNA extracted from formalin-fixed tissue we identified the androgen-upregulated gene ACSL3 as a new 5'-translocation partner of ETV1. These studies report a novel fusion partner for ETV1 and highlight the considerable heterogeneity of ETV1 gene rearrangements in human prostate cancer.
Assuntos
Proteínas de Ligação a DNA/genética , Neoplasias da Próstata/genética , Fatores de Transcrição/genética , Coenzima A Ligases/genética , Estudos de Coortes , Fusão Gênica , Rearranjo Gênico , Heterogeneidade Genética , Humanos , Hibridização in Situ Fluorescente , Masculino , Inclusão em Parafina , Neoplasias da Próstata/patologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Translocação GenéticaRESUMO
Prostate cancer is the most frequently diagnosed male cancer, and its clinical outcome is difficult to predict. The disease may involve the inappropriate expression of genes that normally control the proliferation of epithelial cells in the basal layer and their differentiation into luminal cells. Our aim was to identify novel basal cell markers and assess their prognostic and functional significance in prostate cancer. RNA from basal and luminal cells isolated from benign tissue by immunoguided laser-capture microdissection was subjected to expression profiling. We identified 112 and 267 genes defining basal and luminal populations, respectively. The transcription factor TEAD1 and the ubiquitin ligase c-Cbl were identified as novel basal cell markers. Knockdown of either marker using siRNA in prostate cell lines led to decreased cell growth in PC3 and disrupted acinar formation in a 3D culture system of RWPE1. Analyses of prostate cancer tissue microarray staining established that increased protein levels of either marker were associated with decreased patient survival independent of other clinicopathological metrics. These data are consistent with basal features impacting on the development and clinical course of prostate cancers.
Assuntos
Biomarcadores Tumorais/análise , Proteínas de Ligação a DNA/biossíntese , Proteínas Nucleares/biossíntese , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Proteínas Proto-Oncogênicas c-cbl/biossíntese , Fatores de Transcrição/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Western Blotting , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Imunofluorescência , Expressão Gênica , Perfilação da Expressão Gênica , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Masculino , Microdissecção , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Prognóstico , Neoplasias da Próstata/mortalidade , RNA Interferente Pequeno , Fatores de Transcrição de Domínio TEA , Análise Serial de Tecidos , TransfecçãoRESUMO
Specific chromosomal translocations have now been identified in a variety of solid tumour types. Recent developments in this field include the identification of novel genes involved in the t(X;18) synovial sarcoma translocation and the discovery that the high-mobility group protein gene, HMGI-C, is involved in the chromosome 12 rearrangements found in lipomas and uterine leiomyomas. These discoveries may have important implications for the strategies used in tumour diagnosis and treatment.