RESUMEN
Genomic analysis of tumors is transforming our understanding of cancer. However, although a great deal of attention is paid to the accuracy of the cancer genomic data itself, less attention has been paid to the accuracy of the associated clinical information that renders the genomic data useful for research. In this brief communication, we suggest that omissions and errors in clinical annotations have a major impact on the interpretation of cancer genomic data. We describe our discovery of annotation omissions and errors when reviewing an already carefully annotated colorectal cancer gene expression dataset from our laboratory. The potential importance of clinical annotation omissions and errors was then explored using simulation analyses with an independent genomic dataset. We suggest that the completeness and veracity of clinical annotations accompanying cancer genomic data require renewed focus by the oncology research community, when planning new collections and when interpreting existing cancer genomic data.
Asunto(s)
Genómica , Neoplasias , Humanos , Simulación por Computador , Neoplasias/genéticaRESUMEN
Wilms tumour is a childhood tumour that arises as a consequence of somatic and rare germline mutations, the characterisation of which has refined our understanding of nephrogenesis and carcinogenesis. Here we report that germline loss of function mutations in TRIM28 predispose children to Wilms tumour. Loss of function of this transcriptional co-repressor, which has a role in nephrogenesis, has not previously been associated with cancer. Inactivation of TRIM28, either germline or somatic, occurred through inactivating mutations, loss of heterozygosity or epigenetic silencing. TRIM28-mutated tumours had a monomorphic epithelial histology that is uncommon for Wilms tumour. Critically, these tumours were negative for TRIM28 immunohistochemical staining whereas the epithelial component in normal tissue and other Wilms tumours stained positively. These data, together with a characteristic gene expression profile, suggest that inactivation of TRIM28 provides the molecular basis for defining a previously described subtype of Wilms tumour, that has early age of onset and excellent prognosis.
Asunto(s)
Mutación de Línea Germinal , Neoplasias Renales/genética , Mutación con Pérdida de Función , Recurrencia Local de Neoplasia/genética , Proteína 28 que Contiene Motivos Tripartito/genética , Tumor de Wilms/genética , Adulto , Biomarcadores de Tumor/genética , Epigénesis Genética , Femenino , Perfilación de la Expresión Génica , Humanos , Riñón/patología , Neoplasias Renales/epidemiología , Neoplasias Renales/patología , Masculino , Recurrencia Local de Neoplasia/epidemiología , Recurrencia Local de Neoplasia/patología , Pronóstico , Urotelio/patología , Secuenciación del Exoma , Tumor de Wilms/epidemiología , Tumor de Wilms/patología , Adulto JovenRESUMEN
The decision to authorize a patient for continued enrollment in a state-sanctioned medical cannabis program is difficult in part due to the uncertainty in the accuracy of patient symptom reporting and health functioning including any possible effects on other medication use. We conducted a pragmatic convenience study comparing patient reporting of previous and current prescription opioid usage to the opioid prescription records in the Prescription Monitoring Program (PMP) among 131 chronic pain patients (mean age = 54; 54% male) seeking the first annual renewal of their New Mexico Medical Cannabis Program (NMMCP) license. Seventy-six percent of the patients reported using prescription opioids prior to enrollment in the NMMCP, however, the PMP records showed that only 49% of the patients were actually prescribed opioids in the six months prior to enrollment. Of the 64 patients with verifiable opioid prescriptions prior to NMMCP enrollment, 35 (55%) patients reported having eliminated the use of prescription opioids by the time of license renewal. PMP records showed that 26 patients (63% of patients claiming to have eliminated the use of opioid prescriptions and 41% of all patients with verifiable preenrollment opioid use) showed no prescription opioid activity at their first annual NMMCP renewal visit.
Asunto(s)
Analgésicos Opioides/efectos adversos , Dolor Crónico/tratamiento farmacológico , Marihuana Medicinal/uso terapéutico , Trastornos Relacionados con Opioides/epidemiología , Analgésicos Opioides/orina , Femenino , Humanos , Masculino , Persona de Mediana Edad , New Mexico , Trastornos Relacionados con Opioides/tratamiento farmacológico , Programas de Monitoreo de Medicamentos Recetados/estadística & datos numéricosRESUMEN
BACKGROUND: Co-prescribing of scheduled drugs is endemic in the United Sates, increasing health risks to patients and the burden on healthcare systems. PURPOSE: We conducted a pragmatic historical cohort study to measure the effect of enrollment in a state-authorized United States' Medical Cannabis Program (MCP) on scheduled II-V drug prescription patterns. PROCEDURES: Eighty-three chronic pain patients, who enrolled in the New Mexico MCP between April 1, 2010 and October 3, 2015, were compared with 42 nonenrolled patients over a 24-month period (starting 6 months before enrollment for the MCP patients) using the Prescription Monitoring Program. The outcome variables include baseline levels and pre- and postenrollment monthly trends in the number of drug prescriptions, distinct drug classes, dates prescription drugs were filled, and prescribing providers. FINDINGS: Twenty-eight MCP patients (34%) and 1 comparison group patient (2%) ceased the use of all scheduled prescription medications by the last 6 months of the observation period. Age- and sex-adjusted regressions show that, although no statistically significant differences existed in pre-enrollment levels and trends, the postenrollment trend among MCP patients is statistically significantly negative for all 4 measures (decreases in counts of -0.02 to -0.04, P values between <.001 and .017), whereas the postenrollment trend is 0 among the comparison group. Controlling for time-invariant patient characteristics suggested that MCP patients showed statistically significantly lower levels across all 4 measures by 10 months postenrollment. CONCLUSIONS: Legal access to cannabis may reduce the use of multiple classes of dangerous prescription medications in certain patient populations.
Asunto(s)
Utilización de Medicamentos/legislación & jurisprudencia , Utilización de Medicamentos/estadística & datos numéricos , Fumar Marihuana/epidemiología , Marihuana Medicinal/administración & dosificación , Dolor Intratable/tratamiento farmacológico , Pautas de la Práctica en Medicina/legislación & jurisprudencia , Adulto , Anciano , Dolor Crónico/diagnóstico , Dolor Crónico/tratamiento farmacológico , Estudios de Cohortes , Femenino , Humanos , Incidencia , Masculino , Fumar Marihuana/legislación & jurisprudencia , Persona de Mediana Edad , New Mexico , Medicamentos bajo Prescripción/administración & dosificación , Medición de RiesgoRESUMEN
BACKGROUND: Current levels and dangers of opioid use in the U.S. warrant the investigation of harm-reducing treatment alternatives. PURPOSE: A preliminary, historical, cohort study was used to examine the association between enrollment in the New Mexico Medical Cannabis Program (MCP) and opioid prescription use. METHODS: Thirty-seven habitual opioid using, chronic pain patients (mean age = 54 years; 54% male; 86% chronic back pain) enrolled in the MCP between 4/1/2010 and 10/3/2015 were compared to 29 non-enrolled patients (mean age = 60 years; 69% male; 100% chronic back pain). We used Prescription Monitoring Program opioid records over a 21 month period (first three months prior to enrollment for the MCP patients) to measure cessation (defined as the absence of opioid prescriptions activity during the last three months of observation) and reduction (calculated in average daily intravenous [IV] morphine dosages). MCP patient-reported benefits and side effects of using cannabis one year after enrollment were also collected. RESULTS: By the end of the 21 month observation period, MCP enrollment was associated with 17.27 higher age- and gender-adjusted odds of ceasing opioid prescriptions (CI 1.89 to 157.36, p = 0.012), 5.12 higher odds of reducing daily prescription opioid dosages (CI 1.56 to 16.88, p = 0.007), and a 47 percentage point reduction in daily opioid dosages relative to a mean change of positive 10.4 percentage points in the comparison group (CI -90.68 to -3.59, p = 0.034). The monthly trend in opioid prescriptions over time was negative among MCP patients (-0.64mg IV morphine, CI -1.10 to -0.18, p = 0.008), but not statistically different from zero in the comparison group (0.18mg IV morphine, CI -0.02 to 0.39, p = 0.081). Survey responses indicated improvements in pain reduction, quality of life, social life, activity levels, and concentration, and few side effects from using cannabis one year after enrollment in the MCP (ps<0.001). CONCLUSIONS: The clinically and statistically significant evidence of an association between MCP enrollment and opioid prescription cessation and reductions and improved quality of life warrants further investigations on cannabis as a potential alternative to prescription opioids for treating chronic pain.
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Analgésicos Opioides/uso terapéutico , Dolor Crónico/tratamiento farmacológico , Marihuana Medicinal/uso terapéutico , Estudios de Cohortes , Femenino , Humanos , Masculino , Marihuana Medicinal/efectos adversos , Persona de Mediana Edad , Calidad de VidaRESUMEN
Wilms tumour (WT) is an embryonal tumour that recapitulates kidney development. The normal kidney is formed from two distinct embryological origins: the metanephric mesenchyme (MM) and the ureteric bud (UB). It is generally accepted that WT arises from precursor cells in the MM; however whether UB-equivalent structures participate in tumorigenesis is uncertain. To address the question of the involvement of UB, we assessed 55 Wilms tumours for the molecular features of MM and UB using gene expression profiling, immunohistochemsitry and immunofluorescence. Expression profiling primarily based on the Genitourinary Molecular Anatomy Project data identified molecular signatures of the UB and collecting duct as well as those of the proximal and distal tubules in the triphasic histology group. We performed immunolabeling for fetal kidneys and WTs. We focused on a central epithelial blastema pattern which is the characteristic of triphasic histology characterized by UB-like epithelial structures surrounded by MM and MM-derived epithelial structures, evoking the induction/aggregation phase of the developing kidney. The UB-like epithelial structures and surrounding MM and epithelial structures resembling early glomerular epithelium, proximal and distal tubules showed similar expression patterns to those of the developing kidney. These observations indicate WTs can arise from a precursor cell capable of generating the entire kidney, such as the cells of the intermediate mesoderm from which both the MM and UB are derived. Moreover, this provides an explanation for the variable histological features of mesenchymal to epithelial differentiation seen in WT.
Asunto(s)
Riñón/metabolismo , Mesodermo/metabolismo , Uréter/metabolismo , Tumor de Wilms/genética , Carcinogénesis/genética , Diferenciación Celular/genética , Membrana Celular/genética , Membrana Celular/metabolismo , Feto/metabolismo , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Humanos , Riñón/crecimiento & desarrollo , Riñón/patología , Mesodermo/crecimiento & desarrollo , Técnicas de Cultivo de Órganos , Organogénesis/genética , Uréter/crecimiento & desarrollo , Tumor de Wilms/patologíaRESUMEN
BACKGROUND: Aberrant DNA methylation profiles are a characteristic of all known cancer types, epitomized by the CpG island methylator phenotype (CIMP) in colorectal cancer (CRC). Hypermethylation has been observed at CpG islands throughout the genome, but it is unclear which factors determine whether an individual island becomes methylated in cancer. METHODS: DNA methylation in CRC was analysed using the Illumina HumanMethylation450K array. Differentially methylated loci were identified using Significance Analysis of Microarrays (SAM) and the Wilcoxon Signed Rank (WSR) test. Unsupervised hierarchical clustering was used to identify methylation subtypes in CRC. RESULTS: In this study we characterized the DNA methylation profiles of 94 CRC tissues and their matched normal counterparts. Consistent with previous studies, unsupervized hierarchical clustering of genome-wide methylation data identified three subtypes within the tumour samples, designated CIMP-H, CIMP-L and CIMP-N, that showed high, low and very low methylation levels, respectively. Differential methylation between normal and tumour samples was analysed at the individual CpG level, and at the gene level. The distribution of hypermethylation in CIMP-N tumours showed high inter-tumour variability and appeared to be highly stochastic in nature, whereas CIMP-H tumours exhibited consistent hypermethylation at a subset of genes, in addition to a highly variable background of hypermethylated genes. EYA4, TFPI2 and TLX1 were hypermethylated in more than 90% of all tumours examined. One-hundred thirty-two genes were hypermethylated in 100% of CIMP-H tumours studied and these were highly enriched for functions relating to skeletal system development (Bonferroni adjusted p value =2.88E-15), segment specification (adjusted p value =9.62E-11), embryonic development (adjusted p value =1.52E-04), mesoderm development (adjusted p value =1.14E-20), and ectoderm development (adjusted p value =7.94E-16). CONCLUSIONS: Our genome-wide characterization of DNA methylation in colorectal cancer has identified 132 genes hypermethylated in 100% of CIMP-H samples. Three genes, EYA4, TLX1 and TFPI2 are hypermethylated in >90% of all tumour samples, regardless of CIMP subtype.
Asunto(s)
Adenocarcinoma Mucinoso/genética , Biomarcadores de Tumor/genética , Neoplasias Colorrectales/genética , Islas de CpG , Metilación de ADN , Regulación Neoplásica de la Expresión Génica , Genoma Humano , Adenocarcinoma Mucinoso/patología , Anciano , Neoplasias Colorrectales/patología , Femenino , Humanos , Masculino , Estadificación de Neoplasias , Fenotipo , PronósticoRESUMEN
Epigenetic abnormalities at the IGF2/H19 locus play a key role in the onset of Wilms tumor. These tumors can be classified into three molecular subtypes depending on the events occurring at this locus: loss of imprinting (LOI), loss of heterozygosity (LOH), or retention of imprinting (ROI). As IGF2 LOI is a consequence of aberrant methylation, we hypothesized that this subtype of Wilms tumors might display global abnormalities of methylation. We therefore analyzed the methylation status of satellite DNA, as a surrogate for global methylation in 50 Wilms tumor patients. Satellite methylation was quantified by a methylation-sensitive quantitative PCR. We confirmed hypomethylation of both satellite α (Sat α) and satellite 2 (Sat 2) DNA in Wilms tumor samples compared with normal kidney. In addition, we found that LOI tumors, unlike ROI or LOH ones, showed concordant hypomethylation of both Sat α and Sat 2 DNA. This would suggest that the LOI subtype of Wilms tumor, which unlike other subtypes results from an epimutation, has a global deregulation of methylation mechanisms.
Asunto(s)
Metilación de ADN , ADN Satélite/genética , Impresión Genómica , Factor II del Crecimiento Similar a la Insulina/genética , Tumor de Wilms/genética , Southern Blotting , Inestabilidad Genómica , Humanos , Reacción en Cadena de la Polimerasa , Tumor de Wilms/clasificaciónRESUMEN
Colorectal cancer is one of the five leading causes of cancer mortality worldwide. The mechanisms of pathogen clearance, inflammation and regulation by T cells in the healthy bowel are also important in controlling tumor growth. The majority of studies analyzing T cells and their relationship to colorectal tumor growth have focused on individual T cell markers or gene clusters and thus the complexity of the T cell response contributing to the growth of the tumor is not clear. We have studied the T cells in colorectal cancer patients and have defined a unique T cell signature for colorectal tumor tissue. Using a novel analytical flow cytometric approach in concert with confocal microscopy, we have shown that the tumor has a lower frequency of effector T cells (CD69+), but a higher frequency of both regulatory (CD25hi Foxp3+) and inflammatory T cells (IL-17+) compared with associated nontransformed bowel tissue. We have also identified minor populations of T cells expressing conventional markers of both inflammatory and regulatory T cells (CD4+IL-17+Foxp3+) in the tumor tissue. These cells may represent intermediate populations or they may dictate an inflammatory versus regulatory function in surrounding T cells. Together, these data describe an immune microenvironment in colorectal cancer unique to the tumor tissue and distinct from the surrounding healthy bowel tissue, and this distinct environment is reflected by a gradient of T cells expressing markers of multiple T cell populations. These findings may be used to improve diagnosis and prognosis of colorectal cancer patients.
Asunto(s)
Neoplasias Colorrectales/inmunología , Linfocitos T Reguladores/inmunología , Microambiente Tumoral , Citometría de Flujo , Humanos , Interleucina-17/biosíntesis , Interleucina-17/inmunología , Microscopía ConfocalRESUMEN
BACKGROUND: The immune response has been proposed to be an important factor in determining patient outcome in colorectal cancer (CRC). Previous studies have concentrated on characterizing T cell populations in the primary tumour where T cells with regulatory effect (Foxp3+ Tregs) have been identified as both enhancing and diminishing anti-tumour immune responses. No previous studies have characterized the T cell response in the regional lymph nodes in CRC. METHODS: Immunohistochemistry was used to analyse CD4, CD8 or Foxp3+ T cell populations in the regional lymph nodes of patients with stage II CRC (n = 31), with (n = 13) or without (n = 18) cancer recurrence after 5 years of follow up, to determine if the priming environment for anti-tumour immunity was associated with clinical outcome. RESULTS: The proportions of CD4, CD8 or Foxp3+ cells in the lymph nodes varied widely between and within patients, and there was no association between T cell populations and cancer recurrence or other clinicopathological characteristics. CONCLUSIONS: These data indicate that frequency of these T cell subsets in lymph nodes may not be a useful tool for predicting patient outcome.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Neoplasias Colorrectales/inmunología , Ganglios Linfáticos/inmunología , Recurrencia Local de Neoplasia/inmunología , Subgrupos de Linfocitos T/inmunología , Linfocitos T Reguladores/inmunología , Anciano , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD4-Positivos/patología , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Femenino , Factores de Transcripción Forkhead/metabolismo , Humanos , Técnicas para Inmunoenzimas , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Ganglios Linfáticos/metabolismo , Ganglios Linfáticos/patología , Masculino , Recurrencia Local de Neoplasia/metabolismo , Recurrencia Local de Neoplasia/patología , Estadificación de Neoplasias , Pronóstico , Subgrupos de Linfocitos T/metabolismo , Subgrupos de Linfocitos T/patología , Linfocitos T Reguladores/metabolismo , Linfocitos T Reguladores/patologíaRESUMEN
BACKGROUND: Somatic mutations in the X-linked tumour suppressor gene WTX have been observed in 6- 30% of sporadic cases of Wilms tumour. Germline mutations in the same gene cause the sclerosing skeletal dysplasia, osteopathia striata congenita with cranial sclerosis (OSCS). No evidence points towards a susceptibility to the development of tumours in individuals with OSCS, suggesting that there are unrecognised additional determinants that influence the phenotypic outcome associated with germline mutations in WTX. One explanation may be that a somatic mutation in WTX may need to occur late in tumour development to contribute to tumourigenesis. METHODS: Here a panel of four sporadic Wilms tumours with associated nephrogenic rest tissue and characterised WTX and CTNNB1 mutations is studied to ascertain the temporal sequence of acquisition of these mutations. Additionally, a family with OSCS is described segregating a germline mutation in WTX and manifesting a lethal phenotype in males. One male from this family had bilateral multifocal nephrogenic rests at autopsy. RESULTS: In one of the four tumours the WTX mutation was present in both tumour and rest tissue indicating it had arisen early in tumour development. In the remaining three tumours, the WTX mutation was present in the tumour only indicating late acquisition of these mutations. CONCLUSIONS: These data indicate that WTX mutations can arise both early and late in Wilms tumour development. WTX mutations may predispose to nephrogenic rest development rather than Wilms tumour per se.
Asunto(s)
Mutación , Proteínas Supresoras de Tumor/genética , Tumor de Wilms/genética , Anomalías Múltiples/genética , Anomalías Múltiples/patología , Proteínas Adaptadoras Transductoras de Señales , Secuencia de Bases , Enfermedades del Desarrollo Óseo/patología , Análisis Mutacional de ADN , Salud de la Familia , Resultado Fatal , Femenino , Humanos , Masculino , Osteosclerosis/patología , Linaje , Cráneo/anomalías , Factores de TiempoRESUMEN
Compromised quality of formalin-fixed paraffin-embedded (FFPE)-derived DNA has compounded the use of archival specimens for array-based genomic studies. Recent technological advances have led to first successes in this field; however, there is currently no general agreement on the most suitable platform for the array-based analysis of FFPE DNA. In this study, FFPE and matched fresh-frozen (FF) specimens were separately analyzed with Affymetrix single nucleotide polymorphism (SNP) 6.0 and Agilent 4x44K oligonucleotide arrays to compare the genomic profiles from the two tissue sources and to assess the relative performance of the two platforms on FFPE material. Genomic DNA was extracted from matched FFPE-FF pairs of normal intestinal epithelium from four patients and were applied to the SNP and oligonucleotide platforms according to the manufacturer-recommended protocols. On the Affymetrix platform, a substantial increase in apparent copy number alterations was observed in all FFPE tissues relative to their matched FF counterparts. In contrast, FFPE and matched FF genomic profiles obtained via the Agilent platform were very similar. Both the SNP and the oligonucleotide platform performed comparably on FF material. This study demonstrates that Agilent oligonucleotide array comparative genomic hybridization generates reliable results from FFPE extracted DNA, whereas the Affymetrix SNP-based array seems less suitable for the analysis of FFPE material.
Asunto(s)
Hibridación Genómica Comparativa/métodos , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Adhesión en Parafina , Polimorfismo de Nucleótido Simple , Formaldehído , Dosificación de Gen , Humanos , Mucosa Intestinal , Adhesión en Parafina/métodos , Fijación del TejidoRESUMEN
PURPOSE: The use of adjuvant therapy in stage II colorectal cancer (CRC) remains controversial. There is a need to identify more effective predictors than the traditional staging system to aid therapeutic decision-making. We performed a systematic review and meta-analysis of gene expression profiles (GEPs) to assess their utility for risk stratification and prediction of poor outcomes in stage II CRC. PATIENTS AND METHODS: We performed a comprehensive literature search through December 2007. Studies were included if they reported GEP-based assays in patients with stage II CRC, and either subsequent cancer recurrence or death within 3 years. The prognostic likelihood ratio (LR) and odds ratio (OR) were calculated with 95% confidence intervals and pooled using the fixed-effects method. The weighted average sensitivity, specificity, and accuracy were also reported. RESULTS: Eight cohorts involving 271 patients contributed to the analysis. The average accuracy, sensitivity, and specificity were 81.9%, 76.2%, and 84.5%, respectively, with a prognostic LR of 4.7 (95% CI, 3.2-6.8) and a prognostic OR of 15.1 (95% CI, 7.9-28.6). No evidence for significant interstudy heterogeneity was noted in either analysis. Subgroup analysis found no difference in results for the prediction of cancer recurrence or death. CONCLUSION: This analysis demonstrates the promising potential of using GEP assays as predictors of poor outcomes in stage II CRC, such as cancer recurrence or death. To maximize their utility and availability, further studies will be needed to identify and validate specific gene signatures for poor prognosis in stage II CRC.
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Neoplasias Colorrectales/genética , Perfilación de la Expresión Génica , Antineoplásicos/uso terapéutico , Biomarcadores de Tumor/genética , Neoplasias Colorrectales/patología , ADN de Neoplasias/análisis , Humanos , Estadificación de Neoplasias , Análisis de Secuencia por Matrices de Oligonucleótidos , Resultado del TratamientoRESUMEN
Wilms tumor (WT) is the most frequent renal neoplasm of childhood; a myogenic component is observed in 5% to 10% of tumors. We demonstrate for the first time that myogenic WTs are associated with expression of PAX3, a transcription factor known to specify myoblast cell fate during muscle development. In a panel of 20 WTs, PAX3 was identified in 13 of 13 tumor samples with myogenic histopathology but was absent in 7 of 7 tumors lacking a myogenic component. Furthermore, we show that PAX3 is expressed in the metanephric mesenchyme and stromal compartment of developing mouse kidney. Modulation of endogenous PAX3 expression in human embryonic kidney (HEK293) cells influenced cell migration in in vitro assays. Mutations of WT1 were consistently associated with PAX3 expression in WTs, and modulation of WT1 expression in HEK293 cells was inversely correlated with the level of endogenous PAX3 protein. We demonstrate abundant PAX3 and absence of PAX2 expression in a novel cell line (WitP3) isolated from the stromal portion of a WT bearing a homozygous deletion of the WT1 gene. We hypothesize that PAX3 sets stromal cell fate in developing kidney but is normally suppressed by WT1 during the mesenchyme-to-epithelium transition leading to nephrogenesis. Loss of WT1 permits aberrant PAX3 expression in a subset of WTs with myogenic phenotype.
Asunto(s)
Neoplasias Renales/metabolismo , Riñón/metabolismo , Factores de Transcripción Paired Box/biosíntesis , Tumor de Wilms/metabolismo , Animales , Western Blotting , Diferenciación Celular , Línea Celular Tumoral , Matriz Extracelular/metabolismo , Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Inmunohistoquímica , Riñón/embriología , Neoplasias Renales/genética , Neoplasias Renales/patología , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Mesodermo/metabolismo , Ratones , Factor de Transcripción PAX3 , Factores de Transcripción Paired Box/genética , Fenotipo , Reacción en Cadena de la Polimerasa , ARN Interferente Pequeño , Tumor de Wilms/genética , Tumor de Wilms/patologíaRESUMEN
PURPOSE: The purpose of this study was to identify proteins that are potentially involved in the tumor invasion of bladder cancer. EXPERIMENTAL DESIGN: We searched for the candidate proteins by comparing the profiles of secreted proteins among the poorly invasive human bladder carcinoma cell line RT112 and the highly invasive cell line T24. The proteins isolated from cell culture supernatants were identified by shotgun proteomics. We found that CXCL1 is related to the tumor invasion of bladder cancer cells. We also evaluated whether the amount of the chemokine CXCL1 in the urine would be a potential marker for predicting the existence of invasive bladder tumors. RESULTS: Higher amount of CXCL1 was secreted from highly invasive bladder carcinoma cell lines and this chemokine modulated the invasive ability of those cells in vitro. It was revealed that CXCL1 regulated the expression of matrix metalloproteinase-13 in vitro and higher expression of CXCL1 was associated with higher pathologic stages in bladder cancer in vivo. We also showed that urinary CXCL1 levels were significantly higher in patients with invasive bladder cancer (pT1-4) than those with noninvasive pTa tumors (P = 0.0028) and normal control (P < 0.0001). Finally, it was shown that CXCL1 was an independent factor for predicting the bladder cancer with invasive phenotype. CONCLUSIONS: Our results suggest that CXCL1 modulates the invasive abilities of bladder cancer cells and this chemokine may be a potential candidate of urinary biomarker for invasive bladder cancer and a possible therapeutic target for preventing tumor invasion.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Quimiocina CXCL1/metabolismo , Metaloproteinasa 13 de la Matriz/metabolismo , Neoplasias de la Vejiga Urinaria/metabolismo , Biomarcadores de Tumor/orina , Línea Celular Tumoral , Quimiocina CXCL1/orina , Progresión de la Enfermedad , Humanos , Invasividad Neoplásica , Análisis de Secuencia por Matrices de Oligonucleótidos , Proteómica , Regulación hacia Arriba , Neoplasias de la Vejiga Urinaria/patología , Neoplasias de la Vejiga Urinaria/orinaRESUMEN
Cyclooxygenase-2 (COX-2), the prostaglandin (PG)-synthesizing enzyme overexpressed in colorectal cancer (CRC), has pleiotropic, cancer-promoting effects. COX-2 inhibitors (CIBs) interfere with many cancer-associated processes and show promising antineoplastic activity, however, a common mechanism of CIB action has not yet been established. We therefore investigated by microarray the global response towards the CIB APHS at a dose significantly inhibiting the growth of three COX-2-positive CRC but not of two COX-2-negative cell lines. None of the genes significantly (p = 0.005) affected by APHS were common to all three cell lines and 83% of the altered pathways were cell line-specific. Quantitative polymerase chain reaction (QPCR) on selected pathways confirmed cell line-specific expression alterations induced by APHS. A low stringency data analysis approach using BRB array tools coupled with QPCR, however, identified small expression changes shared by all COX-2-positive cell lines in genes related to the WNT pathway, the key driver of colonic carcinogenesis. Our data indicates a substantial cell line-specificity of APHS-induced expression alterations in CRC cells and helps to explain the divergent effects reported for CIBs. Further, the shared inhibition of the WNT pathway by APHS suggests one potential common mechanism behind the antineoplastic effects of COX-2 inhibition.
RESUMEN
Perilobar (PLNR) and intralobar nephrogenic rests (ILNR) are distinct precursor lesions of Wilms tumors that have different structural, clinical, genetic, and epidemiologic features. Wilms tumors in East-Asian children have unique epidemiologic features in that the incidence is about half that of white children, an early age at diagnosis, a male predominance, and an association with ILNR. Loss of IGF2 imprinting is associated with PLNR more commonly seen in Wilms tumors from white children than tumors from children of Asian descent. Therefore, this epigenetic difference and the higher frequency of PLNR provide an explanation for the interethnic variations in the incidence of Wilms tumor. The histopathologic, clinical, and genetic differences between ILNR and PLNR are described in this review, followed by a description of an epigenetic mechanism that underlies PLNR formation and the unique epidemiologic feature of Wilms tumors.
Asunto(s)
Impresión Genómica , Neoplasias Renales/epidemiología , Neoplasias Renales/genética , Lesiones Precancerosas/epidemiología , Lesiones Precancerosas/genética , Tumor de Wilms/epidemiología , Tumor de Wilms/genética , Pueblo Asiatico , Niño , Humanos , Factor II del Crecimiento Similar a la Insulina , Neoplasias Renales/patología , Masculino , Nefronas/embriología , Nefronas/patología , Lesiones Precancerosas/patología , Proteínas/genética , Tumor de Wilms/patologíaRESUMEN
PURPOSE: It is clinically important to identify bladder cancers with a high risk of intravesical recurrence after transurethral bladder tumor resection. We developed molecular markers for predicting intravesical recurrence of superficial bladder transitional cell carcinoma using oligo-microarray analysis. MATERIALS AND METHODS: Gene expression profiles associated with intravesical recurrence were analyzed by oligo-microarray in 27 superficial bladder transitional cell carcinoma samples from cases treated with transurethral resection between 2000 and 2004 at Kyoto University Hospital. Of candidate genes the expression of P21-activated kinase (Pak1) was validated by semiquantitative real-time polymerase chain reaction using another set of samples and immunohistochemistry. Furthermore, Pak1 functions in bladder cancer cells were analyzed by the transfection of constitutively active (T423E) or kinase dead (K299R) Pak1. RESULTS: Microarray identified 25 genes whose expression was associated with recurrence, including Pak1. Pak1 mRNA expression was statistically associated with grade and the risk of recurrence but not with stage in 86 bladder cancers. Immunohistochemistry and multivariate analysis demonstrated that high Pak1 protein expression was an independent factor associated with recurrence (relative risk 2.27, p = 0.008). High Pak1 expression was significantly associated with a high risk of recurrence even in low stage/grade cancers. Transfection with T423E Pak1 into 253J cells progressed cell motility on wound healing assay, whereas transfection with K299R Pak1 decreased EJ cell motility. CONCLUSIONS: These results suggest that Pak1 expression is associated with recurrence and it might be a useful prognostic marker for superficial bladder transitional cell carcinoma.
Asunto(s)
Biomarcadores de Tumor/análisis , Carcinoma de Células Transicionales/diagnóstico , Recurrencia Local de Neoplasia/diagnóstico , Proteínas Serina-Treonina Quinasas/análisis , Neoplasias de la Vejiga Urinaria/diagnóstico , Anciano , Carcinoma de Células Transicionales/cirugía , Línea Celular Tumoral , Femenino , Perfilación de la Expresión Génica , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias de la Vejiga Urinaria/cirugía , Quinasas p21 ActivadasRESUMEN
The initial development of diffuse gastric cancer (DGC) is poorly understood. The study of E-cadherin (CDH1) germ line mutation carriers predisposed to DGC provides a rare opportunity to elucidate the genetic and biological events surrounding disease initiation. Samples from various stages of hereditary and sporadic DGC were investigated to determine general mechanisms underlying early DGC development. Paraffin-embedded tissues from 13 CDH1 mutation carriers and from 10 sporadic early DGC cases were analyzed. Immunofluorescence and immunohistochemistry using differentiation, proliferation, and adhesion markers showed that DGC initiation seems to occur at the proliferative zone (the upper neck) of the gastric epithelium and correlates with absent or reduced expression of junctional proteins (beta-actin, p120, Lin-7). Slow proliferation of neoplastic cells at the upper gastric neck leads to the formation of intramucosal signet-ring cell carcinoma (SRCC) displaying differentiated features. As shown by immunolabeling, invasion from SRCC lesions beyond the gastric mucosa is associated with poor differentiation, increased proliferation, activation of the c-Src system, and an epithelial-mesenchymal transition. Our results provide a molecular description of the early development of DGC and explain the relationship between the two main DGC types, poorly differentiated carcinoma and SRCC: both share their origin, but SRCC develops following cancer cell differentiation and seems relatively indolent in its intramucosal stage.
Asunto(s)
Proteínas Tirosina Quinasas/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Neoplasias Gástricas/enzimología , Neoplasias Gástricas/patología , Antígenos CD , Proteína Tirosina Quinasa CSK , Cadherinas/genética , Adhesión Celular/fisiología , Diferenciación Celular/fisiología , Procesos de Crecimiento Celular/fisiología , Progresión de la Enfermedad , Activación Enzimática , Mutación de Línea Germinal , Humanos , Neoplasias Gástricas/genética , Familia-src QuinasasRESUMEN
MOTIVATION: The development of gene expression microarray technology has allowed the identification of differentially expressed genes between different clinical phenotypic classes of cancer from a large pool of candidate genes. Although many class comparisons concerned only a single phenotype, simultaneous assessment of the relationship between gene expression and multiple phenotypes would be warranted to better understand the underlying biological structure. RESULTS: We develop a method to select genes related to multiple clinical phenotypes based on a set of multivariate linear regression models. For each gene, we perform model selection based on the doubly-adjusted R-square statistic and use the maximum of this statistic for gene selection. The method can substantially improve the power in gene selection, compared with a conventional method that uses a single model exclusively for gene selection. Application to a bladder cancer study to correlate pre-treatment gene expressions with pathological stage and grade is given. The methods would be useful for screening for genes related to multiple clinical phenotypes. AVAILABILITY: SAS and MATLAB codes are available from author upon request.