RESUMEN
BACKGROUND: Colorectal cancer (CRC) cell lines are widely used pre-clinical model systems. Comprehensive insights into their molecular characteristics may improve model selection for biomedical studies. METHODS: We have performed DNA, RNA and protein profiling of 34 cell lines, including (i) targeted deep sequencing (n = 612 genes) to detect single nucleotide variants and insertions/deletions; (ii) high resolution DNA copy number profiling; (iii) gene expression profiling at exon resolution; (iv) small RNA expression profiling by deep sequencing; and (v) protein expression analysis (n = 297 proteins) by reverse phase protein microarrays. RESULTS: The cell lines were stratified according to the key molecular subtypes of CRC and data were integrated at two or more levels by computational analyses. We confirm that the frequencies and patterns of DNA aberrations are associated with genomic instability phenotypes and that the cell lines recapitulate the genomic profiles of primary carcinomas. Intrinsic expression subgroups are distinct from genomic subtypes, but consistent at the gene-, microRNA- and protein-level and dominated by two distinct clusters; colon-like cell lines characterized by expression of gastro-intestinal differentiation markers and undifferentiated cell lines showing upregulation of epithelial-mesenchymal transition and TGFß signatures. This sample split was concordant with the gene expression-based consensus molecular subtypes of primary tumors. Approximately » of the genes had consistent regulation at the DNA copy number and gene expression level, while expression of gene-protein pairs in general was strongly correlated. Consistent high-level DNA copy number amplification and outlier gene- and protein- expression was found for several oncogenes in individual cell lines, including MYC and ERBB2. CONCLUSIONS: This study expands the view of CRC cell lines as accurate molecular models of primary carcinomas, and we present integrated multi-level molecular data of 34 widely used cell lines in easily accessible formats, providing a resource for preclinical studies in CRC.
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Investigación Biomédica , Neoplasias Colorrectales/metabolismo , Genómica , Proteómica , Secuencia de Bases , Diferenciación Celular , Línea Celular Tumoral , Colon/patología , Neoplasias Colorrectales/genética , Variaciones en el Número de Copia de ADN , Amplificación de Genes , Regulación Neoplásica de la Expresión Génica , Genes Relacionados con las Neoplasias , Inestabilidad Genómica , Humanos , Mutación INDEL/genética , MicroARNs/genética , MicroARNs/metabolismo , Mutación/genética , Fenotipo , Polimorfismo de Nucleótido Simple/genética , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
The prognostic value of CpG island methylator phenotype (CIMP) in colorectal cancer remains unsettled. We aimed to assess the prognostic value of this phenotype analyzing a total of 1126 tumor samples obtained from two Norwegian consecutive colorectal cancer series. CIMP status was determined by analyzing the 5-markers CAGNA1G, IGF2, NEUROG1, RUNX3 and SOCS1 by quantitative methylation specific PCR (qMSP). The effect of CIMP on time to recurrence (TTR) and overall survival (OS) were determined by uni- and multivariate analyses. Subgroup analyses were conducted according to MSI and BRAF mutation status, disease stage, and also age at time of diagnosis (<60, 60-74, ≥75 years). Patients with CIMP positive tumors demonstrated significantly shorter TTR and worse OS compared to those with CIMP negative tumors (multivariate hazard ratio [95% CI] 1.86 [1.31-2.63] and 1.89 [1.34-2.65], respectively). In stratified analyses, CIMP tumors showed significantly worse outcome among patients with microsatellite stable (MSS, P < 0.001), and MSS BRAF mutated tumors (P < 0.001), a finding that persisted in patients with stage II, III or IV disease, and that remained significant in multivariate analysis (P < 0.01). Consistent results were found for all three age groups. To conclude, CIMP is significantly associated with inferior outcome for colorectal cancer patients, and can stratify the poor prognostic patients with MSS BRAF mutated tumors.
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Neoplasias Colorrectales/genética , Islas de CpG/genética , Metilación de ADN/genética , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias Colorrectales/mortalidad , Análisis Mutacional de ADN , Femenino , Humanos , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Mutación , Fenotipo , Reacción en Cadena de la Polimerasa , Modelos de Riesgos Proporcionales , Proteínas Proto-Oncogénicas B-raf/genética , Factores de RiesgoRESUMEN
Colorectal cancer is a global health challenge with high incidence rate and mortality. The patients' prognosis is strongly associated with disease stage and currently there is a need for improved prognostic and predictive biomarkers. In this study, novel colorectal cancer-specific transcript structures were nominated from whole transcriptome sequencing of seven colorectal cancer cell lines, two primary colorectal carcinomas with corresponding normal colonic mucosa and 16 normal tissues. The nominated transcripts were combined with gene level outlier expression analyses in a cohort of 505 colorectal cancers to identify biomarkers with capacity to stratify colorectal cancer subgroups. The transcriptome sequencing data and outlier expression analysis revealed 11 novel colorectal cancer-specific exon-exon junctions, of which 3 were located in the gene VNN1. The junctions within VNN1 were further characterized using rapid amplification of cDNA ends (RACE) and the prevalence of the subsequently characterized novel transcript, VNN1-AB, was investigated by real-time RT-PCR in 291 samples of miscellaneous origins. VNN1-AB was not present in any of the 43 normal colorectal tissue samples investigated, but in 5 of the 6 polyps, and 102 of the 136 (75%) colorectal cancers. We have identified a novel transcript of the VNN1 gene, with an organ-confined complete specificity for colorectal neoplasia.
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Adenoma/genética , Empalme Alternativo/genética , Amidohidrolasas/genética , Biomarcadores de Tumor/genética , Neoplasias Colorrectales/genética , Exones/genética , Adenoma/patología , Colon/metabolismo , Colon/patología , Neoplasias Colorrectales/patología , Proteínas Ligadas a GPI/genética , Perfilación de la Expresión Génica , Humanos , Estadificación de Neoplasias , Análisis de Secuencia por Matrices de Oligonucleótidos , Pronóstico , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
Genes with altered DNA methylation can be used as biomarkers for cancer detection and assessment of prognosis. Here we analyzed the methylation status of a colorectal cancer biomarker panel (CNRIP1, FBN1, INA, MAL, SNCA, and SPG20) in 97 cancer cell lines, derived from 17 different cancer types. Interestingly, the genes were frequently methylated also in hematological cancer types and were therefore subjected to analyses in primary tumor samples from the major types of non-Hodgkin lymphomas (NHL) and in healthy controls. In total, the genes CNRIP1, FBN1, INA, MAL, SNCA, and SPG20 were methylated in 53%, 23%, 52%, 69%, 97%, and 92% of the tumor samples, respectively, and were unmethylated in all healthy controls. With the exception of a single tumor sample, a correct prediction of lymphoma or normal sample was made in a blinded analysis of the validation series using a combination of SNCA and SPG20. The combined ROC-curve analysis of these genes resulted in an area under the curve of 0.999 (P = 4.2 × 10(-18)), and a sensitivity and specificity of 98% and 100%, respectively, across the test and validation series. Interestingly, the promoter methylation of CNRIP1 was associated with decreased overall survival in diffuse large B-cell lymphoma (DLBCL) (P = 0.03). In conclusion, our results demonstrate that SNCA and SPG20 methylation might be suitable for early detection and monitoring of NHL. Furthermore, CNRIP1 could potentially be used as a prognostic factor in DLBCL.
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Biomarcadores de Tumor/genética , Neoplasias Colorrectales/genética , Metilación de ADN , Linfoma no Hodgkin/genética , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/metabolismo , Estudios de Casos y Controles , Proteínas de Ciclo Celular , Línea Celular Tumoral , Neoplasias Colorrectales/metabolismo , Femenino , Humanos , Linfoma no Hodgkin/diagnóstico , Masculino , Persona de Mediana Edad , Pronóstico , Regiones Promotoras Genéticas , Proteínas/genética , alfa-Sinucleína/genéticaRESUMEN
Epigenetic alterations of gene expression are important in the development of cancer. In this study, we identified genes which are epigenetically altered in major lymphoma types. We used DNA microarray technology to assess changes in gene expression after treatment of 11 lymphoma cell lines with epigenetic drugs. We identified 233 genes with upregulated expression in treated cell lines and with downregulated expression in B-cell lymphoma patient samples (nâ=â480) when compared to normal B cells (nâ=â5). The top 30 genes were further analyzed by methylation specific PCR (MSP) in 18 lymphoma cell lines. Seven of the genes were methylated in more than 70% of the cell lines and were further subjected to quantitative MSP in 37 B-cell lymphoma patient samples (diffuse large B-cell lymphoma (activated B-cell like and germinal center B-cell like subtypes), follicular lymphoma and Burkitt`s lymphoma) and normal B lymphocytes from 10 healthy donors. The promoters of DSP, FZD8, KCNH2, and PPP1R14A were methylated in 28%, 67%, 22%, and 78% of the 36 tumor samples, respectively, but not in control samples. Validation using a second series of healthy donor controls (nâ=â42; normal B cells, peripheral blood mononuclear cells, bone marrow, tonsils and follicular hyperplasia) and fresh-frozen lymphoma biopsies (nâ=â25), confirmed the results. The DNA methylation biomarker panel consisting of DSP, FZD8, KCNH2, and PPP1R14A was positive in 89% (54/61) of all lymphomas. Receiver operating characteristic analysis to determine the discriminative power between lymphoma and healthy control samples showed a c-statistic of 0.96, indicating a possible role for the biomarker panel in monitoring of lymphoma patients.
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Linfoma de Células B/genética , Linfoma no Hodgkin/genética , Línea Celular Tumoral , Metilación de ADN/genética , Desmoplaquinas/genética , Canal de Potasio ERG1 , Canales de Potasio Éter-A-Go-Go/genética , Humanos , Péptidos y Proteínas de Señalización Intracelular , Proteínas Musculares , Fosfoproteínas Fosfatasas/genética , Receptores de Superficie Celular/genéticaRESUMEN
The three main types of urological cancers are mostly curable by surgical resection, if early detected. We aimed to identify novel DNA methylation biomarkers common to these three urological cancers, potentially suitable for non-invasive testing. From a candidate list of markers created after gene expression assessment of pharmacologically treated cell lines and tissue samples, two genes were selected for further validation. Methylation levels of these genes were quantified in a total of 12 cancer cell lines and 318 clinical samples. PCDH17 and TCF21 methylation levels provided a sensitivity rate of 92% for bladder cancer, 67% for renal cell tumors and 96% for prostate cancer. Methylation levels were significantly different from those of cancer free individuals (n = 37) for all tumor types (p < 0.001), providing 83% sensitivity and 100% specificity for cancer detection. Although in urine samples the sensitivity was 60%, 32% and 26% for bladder, renal, and prostate tumors, respectively (39% overall), absolute specificity was retained. We identified novel and highly specific methylation markers common to the three main urological cancers. However, additional efforts are required to increase the assay's sensitivity, enabling the simultaneous non-invasive screening of urological tumors in a single voided urine analysis.
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Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Cadherinas/metabolismo , Metilación de ADN/efectos de los fármacos , Neoplasias de la Próstata/diagnóstico , Neoplasias de la Vejiga Urinaria/diagnóstico , Adulto , Anciano , Anciano de 80 o más Años , Azacitidina/análogos & derivados , Azacitidina/farmacología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Biomarcadores/metabolismo , Cadherinas/genética , Carcinoma de Células Renales/diagnóstico , Carcinoma de Células Renales/genética , Carcinoma de Células Renales/metabolismo , Estudios de Casos y Controles , Línea Celular Tumoral , Decitabina , Epigénesis Genética , Femenino , Regulación Neoplásica de la Expresión Génica , Silenciador del Gen , Humanos , Ácidos Hidroxámicos/farmacología , Masculino , Persona de Mediana Edad , Regiones Promotoras Genéticas , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/metabolismo , Curva ROC , Sensibilidad y Especificidad , Neoplasias de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/metabolismoRESUMEN
PURPOSE: To identify a panel of epigenetic biomarkers for accurate bladder cancer (BlCa) detection in urine sediments. EXPERIMENTAL DESIGN: Gene expression microarray analysis of BlCa cell lines treated with 5-aza-2'-deoxycytidine and trichostatin A as well as 26 tissue samples was used to identify a list of novel methylation candidates for BlCa. Methylation levels of candidate genes were quantified in 4 BlCa cell lines, 50 BlCa tissues, 20 normal bladder mucosas (NBM), and urine sediments from 51 BlCa patients and 20 healthy donors, 19 renal cancer patients, and 20 prostate cancer patients. Receiver operator characteristic curve analysis was used to assess the diagnostic performance of the gene panel. RESULTS: GDF15, HSPA2, TMEFF2, and VIM were identified as epigenetic biomarkers for BlCa. The methylation levels were significantly higher in BlCa tissues than in NBM (P < 0.001) and the cancer specificity was retained in urine sediments (P < 0.001). A methylation panel comprising GDF15, TMEFF2, and VIM correctly identified BlCa tissues with 100% sensitivity and specificity. In urine samples, the panel achieved a sensitivity of 94% and specificity of 100% and an area under the curve of 0.975. The gene panel could discriminate BlCa from both healthy individuals and renal or prostate cancer patients (sensitivity, 94%; specificity, 90%). CONCLUSIONS: By using a genome-wide approach, we have identified a biomarker panel that allows for early and accurate noninvasive detection of BlCa using urine samples.
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Biomarcadores de Tumor/genética , Carcinoma/diagnóstico , Epigénesis Genética/genética , Factor 15 de Diferenciación de Crecimiento/genética , Proteínas de la Membrana/genética , Proteínas de Neoplasias/genética , Neoplasias de la Vejiga Urinaria/diagnóstico , Vimentina/genética , Biomarcadores de Tumor/análisis , Carcinoma/genética , Carcinoma/metabolismo , Carcinoma/orina , Estudios de Casos y Controles , Línea Celular Tumoral , ADN de Neoplasias/análisis , ADN de Neoplasias/orina , Detección Precoz del Cáncer/métodos , Perfilación de la Expresión Génica , Factor 15 de Diferenciación de Crecimiento/metabolismo , Factor 15 de Diferenciación de Crecimiento/fisiología , Humanos , Masculino , Proteínas de la Membrana/metabolismo , Proteínas de la Membrana/fisiología , Análisis por Micromatrices , Persona de Mediana Edad , Proteínas de Neoplasias/metabolismo , Proteínas de Neoplasias/fisiología , Pronóstico , Reproducibilidad de los Resultados , Urinálisis/métodos , Neoplasias de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/orina , Vimentina/metabolismo , Vimentina/fisiologíaRESUMEN
PURPOSE: The purpose of this study was to identify genetic aberrations contributing to clinical aggressiveness of malignant peripheral nerve sheath tumors (MPNSTs). PATIENTS AND METHODS: Samples from 48 MPNSTs and 10 neurofibromas were collected from 51 patients with (n = 31) or without (n = 20) neurofibromatosis type 1 (NF1). Genome-wide DNA copy number changes were assessed by chromosomal and array-based comparative genomic hybridization (CGH) and examined for prognostic significance. For a subset of 20 samples, RNA microarray data were integrated with the genome data to identify potential target genes. RESULTS: Forty-four (92%) MPNSTs displayed DNA copy number changes (median, 18 changes per tumor; range, 2 to 35 changes). Known frequent chromosomal gains at chromosome arms 8q (69%), 17q (67%), and 7p (52%) and losses from 9p (50%), 11q (48%), and 17p (44%) were confirmed. Additionally, gains at 16p or losses from 10q or Xq identified a high-risk group with only 11% 10-year disease-specific survival (P = .00005). Multivariate analyses including NF1 status, tumor location, size, grade, sex, complete remission, and initial metastatic status showed that the genomic high-risk group was the most significant predictor of poor survival. Several genes whose expression was affected by the DNA copy number aberrations were identified. CONCLUSION: The presence of specific genetic aberrations was strongly associated with poor survival independent of known clinical risk factors. Conversely, within the total patient cohort with 34% 10-year disease-specific survival, a low-risk group was identified: without changes at chromosomes 10q, 16p, or Xq in their MPNSTs, the patients had 74% 10-year survival.
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Cromosomas Humanos Par 10/genética , Cromosomas Humanos Par 16/genética , Cromosomas Humanos X/genética , Variaciones en el Número de Copia de ADN , Neoplasias de la Vaina del Nervio/genética , Neoplasias del Sistema Nervioso/genética , Adolescente , Adulto , Anciano , Femenino , Expresión Génica , Genoma , Humanos , Masculino , Persona de Mediana Edad , Neoplasias de la Vaina del Nervio/mortalidad , Neoplasias del Sistema Nervioso/mortalidad , Neurofibromatosis 1/genética , Pronóstico , Factores de Riesgo , Análisis de Supervivencia , Adulto JovenRESUMEN
BACKGROUND: Tumor-derived aberrantly methylated DNA might serve as diagnostic biomarkers for cancer, but so far, few such markers have been identified. The aim of the present study was to investigate the potential of the MAL (T-cell differentiation protein) gene as an early epigenetic diagnostic marker for colorectal tumors. METHODS: Using methylation-specific polymerase chain reaction (MSP) the promoter methylation status of MAL was analyzed in 218 samples, including normal mucosa (n = 44), colorectal adenomas (n = 63), carcinomas (n = 65), and various cancer cell lines (n = 46). Direct bisulphite sequencing was performed to confirm the MSP results. MAL gene expression was investigated with real time quantitative analyses before and after epigenetic drug treatment. Immunohistochemical analysis of MAL was done using normal colon mucosa samples (n = 5) and a tissue microarray with 292 colorectal tumors. RESULTS: Bisulphite sequencing revealed that the methylation was unequally distributed within the MAL promoter and by MSP analysis a region close to the transcription start point was shown to be hypermethylated in the majority of colorectal carcinomas (49/61, 80%) as well as in adenomas (45/63, 71%). In contrast, only a minority of the normal mucosa samples displayed hypermethylation (1/23, 4%). The hypermethylation of MAL was significantly associated with reduced or lost gene expression in in vitro models. Furthermore, removal of the methylation re-induced gene expression in colon cancer cell lines. Finally, MAL protein was expressed in epithelial cells of normal colon mucosa, but not in the malignant cells of the same type. CONCLUSION: Promoter hypermethylation of MAL was present in the vast majority of benign and malignant colorectal tumors, and only rarely in normal mucosa, which makes it suitable as a diagnostic marker for early colorectal tumorigenesis.
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Biomarcadores de Tumor/metabolismo , Neoplasias Colorrectales/diagnóstico , Neoplasias Colorrectales/genética , Metilación de ADN , Proteínas de Transporte de Membrana/genética , Proteínas de la Mielina/genética , Proteolípidos/genética , Neoplasias Colorrectales/metabolismo , Humanos , Proteínas Proteolipídicas Asociadas a Mielina y Linfocito , Regiones Promotoras GenéticasRESUMEN
Molecular markers that could stratify prostate cancer patients according to risk of disease progression would allow a significant improvement in the management of this clinically heterogeneous disease. In the present study, we analyzed the genetic profile of a consecutive series of 51 clinically confined prostate carcinomas and 27 benign prostatic hyperplasias using comparative genomic hybridization (CGH). We then added our findings to the existing literature data in order to perform a meta-analysis on a total of 294 prostate cancers with detailed CGH and clinicopathological information, using multivariate statistical methods that included principal component, hierarchical clustering, time of occurrence, and regression analyses. Whereas several genomic imbalances were shared by organ-confined, locally invasive, and metastatic prostate cancers, 6q and 10q losses and 7q and 8q gains were significantly more frequent in patients with extra-prostatic disease. Regression analysis indicated that 8q gain and 13q loss were the best predictors of locally invasive disease, whereas 8q gain and 6q and 10q losses were associated with metastatic disease. We propose a genetic pathway of prostate carcinogenesis with two distinct initiating events, namely, 8p and 13q losses. These primary imbalances are then preferentially followed by 8q gain and 6q, 16q, and 18q losses, which in turn are followed by a set of late events that make recurrent and metastatic prostate cancers genetically more complex. We conclude that significant differences exist in the genetic profile of organ-confined, locally invasive, and advanced prostate cancer and that genetic features may carry prognostic information independently of Gleason grade.
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Neoplasias de la Próstata/genética , Anciano , Humanos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Hibridación de Ácido Nucleico , Hiperplasia Prostática/genéticaRESUMEN
It was recently shown by flow and static cytometry that a large sub-group of follicular adenomas of the thyroid--fetal/embryonal adenomas--display an aneuploid phenotype. It was also shown that thyroid lesions with a DNA content within the triploid range were either fetal adenomas or follicular carcinomas with a fetal adenoma growth pattern. Follicular tumours with growth patterns other than the so-called fetal adenoma-like pattern were usually diploid or near-diploid. In an attempt to clarify the pattern of chromosomal imbalances in follicular tumours, comparative genomic hybridization (CGH) analysis was performed in a series of 18 follicular neoplasms (ten fetal/embryonal and four common follicular adenomas and four minimally invasive follicular carcinomas). For each tumour, the DNA content was determined by flow cytometry and, in some cases, also by static cytometry. Finally, the copy number of selected chromosomes was determined by interphase fluorescence in situ hybridization (FISH) using centromere probes. With the exception of the single diploid fetal adenoma, all fetal adenomas displayed several DNA copy number changes, with frequent gains of several chromosomes, which were found to be either tetrasomic or trisomic by FISH. This genetic pattern was also present in the single case of follicular carcinoma with aneuploidy and fetal adenoma-like growth pattern. Follicular adenomas other than fetal adenomas, and the remaining follicular carcinomas, showed more losses than gains of chromosomes. These results suggest that follicular tumourigenesis may follow at least two pathways: one characterized by prominent aneuploidy and numerous gains, in which the tumours display a fetal adenoma-like growth pattern; and another accompanied by less obvious aneuploidy or even quasi-diploidy and dominant chromosome losses, in which the tumours display a common follicular architecture.
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Adenoma/genética , Aberraciones Cromosómicas , Neoplasias de la Tiroides/genética , Adenoma/patología , Aneuploidia , ADN de Neoplasias/genética , Diploidia , Enfermedades Fetales/genética , Enfermedades Fetales/patología , Citometría de Flujo/métodos , Humanos , Hibridación Fluorescente in Situ/métodos , Invasividad Neoplásica , Hibridación de Ácido Nucleico/métodos , Glándula Tiroides/patología , Neoplasias de la Tiroides/patologíaRESUMEN
In cancer biology, cell lines are often used instead of primary tumors because of their widespread availability and close reflection of the in vivo state. Cancer is a genetic disease, commonly caused by small- and large-scale DNA rearrangements. Therefore, it is essential to know the genomic profiles of tumor cell lines to enable their correct and efficient use as experimental tools. Here, we present a comprehensive study of the genomic profiles of 20 colon cancer cell lines combining conventional karyotyping (G-banding), comparative genomic hybridization (CGH), and multicolor fluorescence in situ hybridization (M-FISH). Major differences between the microsatellite instability (MSI) and chromosome instability (CIN) cell lines are shown; the CIN cell lines exhibited complex karyotypes involving many chromosomes (mean: 8.5 copy number changes), whereas the MSI cell lines showed considerably fewer aberrations (mean: 2.6). The 3 techniques complement each other to provide a detailed picture of the numerical and structural chromosomal changes that characterize cancer cells. Therefore, 7 of the cell lines (Colo320, EB, Fri, IS2, IS3, SW480, and V9P) are here completely karyotyped for the first time and, among these, 5 have not previously been cytogenetically described. By hierarchical cluster analysis, we show that the cell lines are representative models for primary carcinomas at the genome level. We also present the genomic profiles of an experimental model for tumor progression, including 3 cell lines (IS1, IS2, and IS3) established from a primary carcinoma, its corresponding liver- and peritoneal metastasis from the same patient. To address the question of clonality, we compared the genome of 3 common cell lines grown in 2 laboratories. Finally, we compared all our results with previously published CGH data and karyotypes of colorectal cell lines. In conclusion, the large variation in genetic complexity of the cell lines highlights the importance of a comprehensive reference of genomic profiles for investigators engaged in functional studies using these research tools.
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Neoplasias del Colon/genética , Neoplasias del Colon/metabolismo , Genoma Humano , Línea Celular Tumoral , Inestabilidad Cromosómica , Aberraciones Cromosómicas , Bandeo Cromosómico , Células Clonales , Análisis por Conglomerados , Neoplasias del Colon/patología , Perfilación de la Expresión Génica , Variación Genética , Humanos , Hibridación Fluorescente in Situ , Cariotipificación , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/secundario , Repeticiones de Microsatélite , Metástasis de la Neoplasia , Hibridación de Ácido Nucleico , Neoplasias Peritoneales/genética , Neoplasias Peritoneales/secundario , PloidiasRESUMEN
BACKGROUND: The widespread use of prostate-specific antigen (PSA) testing to screen for prostate carcinoma has led to significant overdiagnosis, due to the frequent detection of indolent malignancies on PSA screening. The detection of abnormal PSA levels typically is followed by ultrasound-guided needle biopsy. Therefore, in an effort to identify genetic markers that augment the information provided by standard histopathologic classification, the authors tested the feasibility of using these minute biopsy samples for genomic profiling via chromosome banding analysis and comparative genomic hybridization (CGH). METHODS: Ultrasound-guided needle biopsy specimens obtained preoperatively from 35 patients with prostate carcinoma were analyzed via chromosome banding analysis (after short-term culturing) and CGH. The findings of these analyses then were analyzed for potential correlations with clinicopathologic parameters. RESULTS: Chromosome banding analysis and CGH were possible in 34 and 33 of the 35 study specimens, respectively. Combined analysis revealed aberrations in 69% of all samples investigated. Copy number losses occurred most commonly at 8p (58% of all abnormal specimens), 16q (42%), and 13q (37%), whereas the only gains detected in more than 1 specimen were those that occurred at 8q (37%). Genomic imbalances and losses at 16q were significantly associated with more poorly differentiated subtypes of prostate carcinoma (P = 0.048 and P = 0.019, respectively), whereas gains at 8q and losses at 16q were significantly correlated with clinically advanced disease (P = 0.048 for the finding of a gain at 8q together with a loss at 16q; P = 0.01 for the finding of either aberration alone). CONCLUSIONS: The authors conclude that genomic analysis of suspected prostate carcinoma specimens obtained via ultrasound-guided needle biopsy is feasible. Thus, it may be possible to use genetic markers to obtain diagnostic and/or prognostic information that is useful in the making of preoperative decisions regarding prostate carcinoma management.
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Aberraciones Cromosómicas , Neoplasia Intraepitelial Prostática/diagnóstico , Neoplasia Intraepitelial Prostática/genética , Neoplasias de la Próstata/diagnóstico , Neoplasias de la Próstata/genética , Anciano , Biopsia , Diferenciación Celular , Bandeo Cromosómico , Cromosomas Humanos Par 16/genética , ADN de Neoplasias/genética , ADN de Neoplasias/metabolismo , Toma de Decisiones , Estudios de Factibilidad , Humanos , Cariotipificación , Masculino , Persona de Mediana Edad , Hibridación de Ácido Nucleico , Cuidados Preoperatorios , Pronóstico , Antígeno Prostático Específico/metabolismoRESUMEN
BACKGROUND: Colorectal cancer (CRC) is one of the most common causes of cancer-related deaths in the Western world, and despite the fact that metastases are usually the ultimate cause of deaths, the knowledge of the genetics of advanced stages of this disease is limited. In order to identify potential genetic abnormalities underlying the development of local and distant metastases in CRC patients, we have, by comparative genomic hybridization, compared the DNA copy number profiles of 10 primary carcinomas, 14 local recurrences, 7 peritoneal carcinomatoses, and 42 liver metastases from 61 CRC patients. RESULTS: The median number of aberrations among the primary carcinomas, local recurrences, carcinomatoses, and liver metastases was 10, 6, 13, and 14, respectively. Several genetic imbalances, such as gains of 7, 8q, 13q, and 20, and losses of 4q, 8p, 17p, and 18, were common in all groups. In contrast, gains of 5p and 12p were more common in the carcinomatoses than in other stages of the disease. With hierarchical cluster analysis, liver metastases could be divided into two main subgroups according to clusters of chromosome changes. CONCLUSIONS: Each stage of CRC progression is characterized by a particular genetic profile, and both carcinomatoses and liver metastases are more genetically complex than local recurrences and primary carcinomas. This is the first genome profiling of local recurrences and carcinomatoses, and gains of 5p and 12p seem to be particularly important for the spread of the CRC cells within the peritoneal cavity.
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Carcinoma/patología , Neoplasias Colorrectales/patología , Genoma Humano , Neoplasias Hepáticas/secundario , Carcinoma/genética , Aberraciones Cromosómicas , Análisis por Conglomerados , Neoplasias Colorrectales/genética , Humanos , Neoplasias Hepáticas/genética , Recurrencia Local de Neoplasia , Estadificación de Neoplasias , Hibridación de Ácido Nucleico/métodosRESUMEN
Desmoid tumors are benign neoplasms of the fibromatosis group. Data on their acquired chromosomal changes are sparse and, therefore, we wanted to ascertain what genomic losses and gains these tumors may have incurred. DNA was extracted from a total of 26 formalin-fixed, paraffin-embedded desmoid tumors followed by comparative genomic hybridization (CGH) and interphase fluorescence in situ hybridization (I-FISH) analyses. Ten of 12 informative tumors were normal by CGH; the two abnormal ones had loss of chromosome 6 and loss of 6q and gain of chromosome 20, respectively. I-FISH analyses with an alpha-satellite probe specific for chromosome 8 of 26 desmoids, including one tumor that by karyotyping had +i(8)(q10), showed no evident abnormalities. An explanation for the relatively high frequency of genomically normal tumors by CGH seen in this study may be sought in the fact that as many as 10 of the 12 informative tumors were abdominal desmoids, a subset of tumors also previously found to exhibit genomic changes only rarely. It is therefore possible that abdominal desmoids might be non-neoplastic tumors or neoplastic tumors with genetic changes too small to be discovered by CGH, whereas desmoid tumors from other locations exhibit detectable genomic changes at a significantly higher frequency.
Asunto(s)
Cromosomas Humanos Par 8 , Fibromatosis Abdominal/genética , Adulto , Anciano , Femenino , Humanos , Hibridación Fluorescente in Situ , Cariotipificación , Masculino , Persona de Mediana Edad , Hibridación de Ácido NucleicoRESUMEN
The majority of genetic studies of colorectal carcinogenesis have focused on changes found in primary tumors. Despite the fact that liver metastases are a leading cause of colorectal cancer deaths, the molecular genetic basis of the advanced disease stages remains poorly understood. We performed comparative genomic hybridization (CGH) on 17 liver metastases from colorectal carcinomas and compared the quantitative profile with the qualitative profile previously obtained with chromosome banding. An average of 12.6 aberrations per tumor was found by CGH. Chromosome 18 and chromosome arms 4q, 8p, and 17p were most frequently lost, whereas chromosomes 7 and 20 and chromosome arms 6p, 8q, and 13q were most frequently gained. We compared the chromosome banding and CGH data after converting the karyotypes into net copy number gains and losses. Ten tumors showed agreement between the findings of the two techniques, whereas five tumors did not (in two cases, no mitotic cells were obtained for banding analysis). All five discordant cases had a "simple" abnormal or normal karyotype, but revealed multiple changes by CGH. A likely explanation for this discrepancy is that in vitro growth before G-banding selected against the cancer cells. Interestingly, by comparing the CGH profiles of the "complex" vs. the "simple"/normal karyotype groups, deletion of 8p and gain of 16q were seen more frequently in the former group. The liver metastases had the same aberrations as seen in primary colorectal carcinomas, summarized in a literature survey. However, these aberrations were seen more frequently in liver metastases, which may be attributable to increased genetic instability.
Asunto(s)
Bandeo Cromosómico , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Análisis Citogenético/métodos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/secundario , Aberraciones Cromosómicas , Bandeo Cromosómico/métodos , Deleción Cromosómica , Femenino , Amplificación de Genes/genética , Humanos , Cariotipificación/métodos , Masculino , Hibridación de Ácido Nucleico/métodosRESUMEN
In order to investigate the genetics of testicular germ cell tumors (TGCTs), we examined 33 TGCTs, including 15 familial/bilateral and 18 sporadic tumors, using comparative genomic hybridization. The frequencies of the histological subtypes were comparable between the two groups. Gains of the whole or parts of chromosome 12 were found in 30 tumors (91%). Furthermore, increased copy number of the whole or parts of chromosomes 7, 8, 17, and X, and decreased copy number of the whole or parts of chromosomes 4, 11, 13, and 18 were observed in > or = 50% of the tumors. Sixteen smallest regions of overlapping changes were delineated on 12 different chromosomes. The chromosomal copy numbers of familial/bilateral and sporadic TGCTs were comparable, suggesting similar genetic pathways to disease in both groups. However, significant differences were observed between the two main histological subgroups. Gains from 15q and 22q were associated with seminomas (P = 0.005 and P = 0.02, respectively), whereas gain of the proximal 17q (17q11.2-21) and high-level amplification from chromosome arm 12p, and losses from 10q were associated with nonseminomas (P < 0.001, P = 0.04, and P = 0.03, respectively).