RESUMEN
Interleukin-1 beta (IL-1 beta) mediates a wide range of immune and inflammatory responses. The active cytokine is generated by proteolytic cleavage of an inactive precursor. A complementary DNA encoding a protease that carries out this cleavage has been cloned. Recombinant expression in COS-7 cells enabled the cells to process precursor IL-1 beta to the mature form. Sequence analysis indicated that the enzyme itself may undergo proteolytic processing. The gene encoding the protease was mapped to chromosomal band 11q23, a site frequently involved in rearrangement in human cancers.
Asunto(s)
Cromosomas Humanos Par 11 , Precursores Enzimáticos/genética , Metaloendopeptidasas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Caspasa 1 , Línea Celular , Bandeo Cromosómico , Clonación Molecular , Precursores Enzimáticos/biosíntesis , Precursores Enzimáticos/aislamiento & purificación , Humanos , Metaloendopeptidasas/biosíntesis , Metaloendopeptidasas/aislamiento & purificación , Datos de Secuencia Molecular , Neutrófilos/enzimología , Oligodesoxirribonucleótidos , Poli A/genética , Poli A/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , ARN/genética , ARN/aislamiento & purificación , ARN Mensajero/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/aislamiento & purificación , TransfecciónRESUMEN
The human chromosomal location of the gene encoding the phosphatidylinositol-3 kinase associated protein, p85 alpha, has been determined by analysis of its segregation in rodent-human hybrids and by chromosome in situ hybridization using a complementary DNA clone, GRB-1. The gene for p85 alpha is at chromosome region 5q13, perhaps near the gene encoding another receptor associated signal transducing protein, the GTPase activating protein.
Asunto(s)
Mapeo Cromosómico , Cromosomas Humanos Par 5 , Genoma Humano , Fosfotransferasas/genética , 1-Fosfatidilinositol 4-Quinasa , Deleción Cromosómica , HumanosRESUMEN
Extensive studies of loss of heterozygosity of 3p markers in renal cell carcinomas (RCCs) have established that there are at least three regions critical in kidney tumorigenesis, one most likely coincident with the von Hippel-Lindau gene at 3p25.3, one in 3p21 which may also be critical in small cell lung carcinomas, and one in 3p13-p14.2, a region which includes the 3p chromosome translocation break of familial RCC with the t(3;8)(p14.2;q24.1) translocation. A panel of rodent-human hybrids carrying portions of 3p, including a hybrid carrying the derivative 8 (der(8)(8pter-->8q24.1::3p14.2-->3pter)) from the RCC family, have been characterized using 3p anchor probes and cytogenetic methods. This 3p panel was then used to map a large number of genetically mapped probes into seven physical intervals between 3p12 and 3pter defined by the hybrid panel. Markers have been physically, and some genetically, placed relative to the t(3;8) break, such that positional cloning of the break is feasible.
Asunto(s)
Carcinoma de Células Renales/genética , Cromosomas Humanos Par 3 , Cromosomas Humanos Par 8 , Neoplasias Renales/genética , Translocación Genética , Animales , Deleción Cromosómica , Mapeo Cromosómico , Cricetinae , Sondas de ADN , ADN de Neoplasias/genética , Heterocigoto , Humanos , Células Híbridas/fisiología , Hibridación Fluorescente in Situ , MetafaseRESUMEN
Chromosomal region 11q23 participates in a number of reciprocal translocations with specific regions of chromosomes 4, 9, 19, and others. These translocations are associated with acute lymphocytic leukemia and acute myelomonocytic, monocytic, and myelogenous leukemia. From a yeast artificial chromosome containing human DNA derived from 11q23 we cloned a DNA fragment which can be used as a probe to detect rearrangements in leukemic cells from the majority of patients with the t(4;11), t(9;11), and t(11;19) translocations. The breakpoints cluster in a small DNA region of less than 5.8 kilobases.
Asunto(s)
Cromosomas Humanos Par 11 , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Translocación Genética/genética , Antígenos de Diferenciación de Linfocitos T/genética , Northern Blotting , Southern Blotting , Complejo CD3 , Mapeo Cromosómico , Cromosomas Humanos Par 19 , Cromosomas Humanos Par 4 , Cromosomas Humanos Par 9 , Clonación Molecular , ADN de Neoplasias/genética , Humanos , ARN Mensajero/genética , ARN Neoplásico/genética , Receptores de Antígenos de Linfocitos T/genética , Mapeo RestrictivoRESUMEN
The B-raf gene was localized to human chromosome region 7p11-7qter by Southern blot analysis of its segregation in a panel of rodent-human hybrids, using a B-raf cDNA clone. Chromosomal in situ hybridization refined the localization to 7q33-36. Analysis of genomic B-raf clones identified a B-raf pseudogene in addition to the active gene. Based on the chromosomal mapping data we conclude that the pseudogene is located near the active gene.
Asunto(s)
Cromosomas Humanos Par 7 , Proteínas Proto-Oncogénicas/genética , Secuencia de Bases , Southern Blotting , Mapeo Cromosómico , Humanos , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Proteínas Quinasas/genética , Proteínas Proto-Oncogénicas c-raf , Proto-Oncogenes , Seudogenes , Mapeo RestrictivoRESUMEN
FHIT (Fragile Histidine Triad), a putative tumor suppressor gene, was cloned from fetal brain and colon cDNA libraries. Portions of this gene are deleted in esophageal, colon, lung and breast tumors, but this gene has not been found altered in sporadic renal cell carcinomas. We report here an alternatively spliced form of this gene cloned from a kidney cDNA library. This cDNA is 1189 bp in length, and contains an additional 94 bp exon, designated exon 2a (E2a). This novel sequence is located between exon 2 and exon 3 of the FHIT gene's untranslated region and exon 2a is present in all normal kidney tissues and cell lines. Analyses performed on sporadic renal cell carcinoma (RCC) tissues and cell lines, show consistent loss of exon 8 of the FHIT cDNA in almost 60% of the cases. Interestingly, in a familial, as well as, in a metastatic RCC, derived from a patient with the sporadic form, exon 2a and exon 3 are also deleted. Northern analyses with the exon 2a of the familial and the metastatic RCC demonstrates concurrent loss of expression of a 4.4 kb transcript with the loss of the E2a sequence, suggesting that exon 2a of the FHIT gene may play an important role in the oncogenesis of renal cell carcinoma.
Asunto(s)
Ácido Anhídrido Hidrolasas , Empalme Alternativo , Carcinoma de Células Renales/genética , Genes Supresores de Tumor , Neoplasias Renales/genética , Proteínas de Neoplasias/genética , Proteínas/genética , Secuencia de Aminoácidos , Secuencia de Bases , Northern Blotting , Clonación Molecular , Humanos , Riñón , Datos de Secuencia Molecular , Reacción en Cadena de la PolimerasaRESUMEN
The transfection recombinant transforming gene, tre, originated from discontinuous human genetic elements after transfection of NIH3T3 cells with genomic DNA from a Ewing's sarcoma cell line. Probes for the three normally discontinuous human elements involved in the transfection recombinant were subcloned and used in conjunction with a panel of rodent-human hybrid cells to determine their normal location in the human genome. The leftmost (5') element derives from the long arm of chromosome 5 (5q); the internal fragment derives from human chromosome 18 proximal to the bcl-2 gene; and the rightmost (3') element derives from the long arm of chromosome 17 (17q) distal to an acute leukemia breakpoint at 17q21. In situ hybridization of the same probes to human metaphase chromosomes confirmed localization of these sequences to regions 5q23----q31, 18q12, and 17q12----q22.
Asunto(s)
Cromosomas Humanos Par 17 , Cromosomas Humanos Par 18 , Cromosomas Humanos Par 5 , Reordenamiento Génico , Oncogenes , Animales , Bandeo Cromosómico , Mapeo Cromosómico , Sondas de ADN , Humanos , Células Híbridas/citología , Hibridación de Ácido Nucleico , Mapeo RestrictivoRESUMEN
Tre is a recombinant gene isolated from NIH3T3 cells transfected with human Ewing's sarcoma DNA. It is composed of three major genetic elements derived, 5' to 3', from human chromosomes 5, 18 and 17. We report here on transcripts from the 3' domain of tre. The transcripts were cloned from a cDNA library of cytoplasmic poly(A)+ RNA from tre-transfected NIH3T3 tumor cells. The complete cDNA sequence, 8201 nucleotides, possessed an unusually long non-coding region and a translatable region with two open reading frames. In one cDNA clone, the presence of two insertions suggested the possibility of alternative splicing. The sequence mapped to the centromere-proximal region of 17q. Transfection-tumorigenicity assays with the open reading frames subcloned into expression vectors were positive for the reading frame adjacent to the 5' non-coding region and negative for the second, downstream, reading frame and the possible alternatively spliced versions of both reading frames. Analysis of the 786 amino acid sequence deduced from the 5' reading frame predicted a highly hydrophilic protein with two charge clusters suggesting nucleic acid-binding properties. When used as probe, the cloned sequence detected RNA transcripts in a wide variety of human cancer cells regardless of their lineage of origin from different tissues, but not in human cells from normal tissue.
Asunto(s)
Endopeptidasas , Proteínas de Fusión Oncogénica/genética , Proteínas Oncogénicas , Proteínas Proto-Oncogénicas/genética , Proto-Oncogenes , Células 3T3 , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cromosomas Humanos Par 17 , Clonación Molecular , ADN/genética , Expresión Génica , Humanos , Técnicas In Vitro , Ratones , Datos de Secuencia Molecular , ARN Mensajero/genética , Mapeo Restrictivo , Transcripción Genética , Células Tumorales Cultivadas , Ubiquitina TiolesterasaRESUMEN
AP-1060 is a newly established acute promyelocytic leukemia (APL) cell line from a multiple-relapse patient clinically resistant to both all-trans retinoic acid (ATRA) and arsenic trioxide (ATO). The line was initially derived as a granulocyte colony-stimulating factor-dependent strain that underwent replicative senescence and, following ethylnitrosourea treatment, as a phenotypically similar immortalized line. Immortalization was associated with broadened cytokine sensitivity but not growth autonomy, in contrast to three previously derived APL lines. Both the AP-1060 strain and line had shortened telomeres and low telomerase activity, while the line had higher expression of many genes associated with macromolecular synthesis. The karyotype was 46,XY,t(3;14)(p21.1;q11.2),t(15;17)(q22;q11)[100%]; the unique t(3;14) was observed in 4/9 t(15;17)-positive metaphase cells at previous relapse on ATRA therapy. The PML-RARalpha mRNA harbored a missense mutation in the RARalpha-region ligand-binding domain (Pro900Ser). This was associated with a right-shift and sharpening of the ATRA-induced maturation response compared to ATRA-sensitive NB4 cells, which corresponded to the transcriptional activation by PML-RARalphaPro900Ser of a cotransfected ATRA-targeted reporter vector in COS-1 cells. AP-1060 also manifested relative resistance to ATO-induced apoptosis at >/=1 microM, while 0.25 microM ATO stimulated limited atypical maturation. These findings suggest that AP-1060 will be useful for further assessing molecular elements involved in APL progression and drug response/resistance.
Asunto(s)
Línea Celular Tumoral , Resistencia a Antineoplásicos , Leucemia Promielocítica Aguda/patología , Trióxido de Arsénico , Arsenicales/farmacología , Técnicas de Cultivo de Célula/métodos , Citocinas/farmacología , Humanos , Cariotipificación , Mutación Missense , Óxidos/farmacología , Receptores de Ácido Retinoico/genética , Receptor alfa de Ácido Retinoico , Telomerasa/metabolismo , Telómero/ultraestructura , Tretinoina/farmacologíaRESUMEN
In order to contribute to the study of the molecular basis of leukemic cellular resistance to the induction of differentiation by all-trans retinoic acid (RA) we have generated and analyzed a mutant, RA-resistant HL-60 cell line. Molecular analysis of the retinoic acid receptor alpha (RARalpha) cDNA disclosed, in one of the two alleles, a novel mutation consisting of a 7-base deletion in the ligand binding domain that includes part of a FokI restriction endonuclease site previously described. As a consequence of this deletion and translational frame-shift, a stop signal is created that truncates the protein at codon 421, disrupting an essential functional component of the receptor. Transducing an epitope tagged RARalpha into the mutant is sufficient to inhibit clonal growth in the presence of RA. Standard cytogenetic analysis, fluorescent in situ hybridization (FISH) and comparative genomic hybridization (CGH) analysis revealed the presence of two RARalpha loci, and showed a composite karyotype with additional abnormalities with respect to the parental line, including a chromosome 8 insertion in a chromosome previously known as marker three.
Asunto(s)
Aberraciones Cromosómicas , Mutación , Receptores de Ácido Retinoico/genética , Tretinoina/farmacología , Alelos , Cromosomas Humanos Par 8 , Resistencia a Antineoplásicos , Marcadores Genéticos , Células HL-60 , Humanos , Hibridación Fluorescente in Situ , Receptor alfa de Ácido RetinoicoRESUMEN
One of the most important biological features of papillary transitional cell carcinoma (pTCC) of the urinary tract is its multicentricity and its tendency for recurrences. Two possible mechanisms, field effect and intramucosal seeding/spreading, have been proposed. The former theory hypothesizes that carcinogenic agents cause synchronous or metachronous malignant transformation of multiple urothelial cells (independent clonal origin), and the latter speculates that synchronous and metachronous tumors are derived from implantation or direct spreading of tumor cells (identical clonal origin). We tested these hypotheses by analyzing the methylation patterns of the androgen receptor gene (HUMARA) located at the X-chromosome. Thirty-five metachronous and synchronous, low-grade (grade 1 or 2), noninvasive pTCCs of the urinary tract from 10 heterozygous female patients were successfully analyzed using formalin-fixed, paraffin-embedded tissue. These included 16 recurrent bladder tumors from 4 patients, 10 metachronous bladder and ureter/renal pelvis tumors from 4 patients, and 9 multifocal tumors from 2 patients. All tumors are monoclonal as indicated by unbalanced methylation of HUMARA. Furthermore, same methylated allele was detected in multiple recurrent or multifocal tumors from any given patient, indicating their identical clonal origin. We conclude that low-grade, noninvasive pTCCs are monoclonal in nature. Synchronous or metachronous pTCCs have an identical clonal origin, strongly supporting the intramucosal seeding/spreading hypothesis.
Asunto(s)
Carcinoma Papilar/patología , Carcinoma de Células Transicionales/patología , Neoplasias Primarias Múltiples/patología , Neoplasias Primarias Secundarias/patología , Neoplasias Urológicas/patología , Carcinoma Papilar/genética , Carcinoma de Células Transicionales/genética , Células Clonales , Femenino , Humanos , Neoplasias Primarias Múltiples/genética , Neoplasias Primarias Secundarias/genética , Neoplasias Urológicas/genéticaRESUMEN
Endometrial hyperplasia is a common disorder that is now observed with increasing frequency in women treated with hormonal replacement therapy or with tamoxifen. This study was undertaken to determine whether genomic features of various forms of endometrial hyperplasias would allow their classification as a benign, premalignant, or malignant abnormality. Comparative genomic hybridization (CGH) was performed on endometrial glands microdissected by laser capture microscope from 19 archival endometrial samples, comprising 5 normal endometria, 1 polyp, 2 simple hyperplasias, 5 hyperplasias with nuclear abnormalities (atypical hyperplasias), and 4 low-grade and 2 high-grade endometrioid carcinomas, 1 with squamous component (adenoacanthoma). Genomic DNA, extracted from the glands and the squamous component in 1 case, was amplified by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR) and compared with sex-matched DNA by CGH. No genomic imbalances were observed in the normal samples, the polyp, or the simple hyperplasias. However, in atypical hyperplasia, regardless of the level of cytologic atypia, genomic abnormalities were observed that also occurred in endometrioid carcinomas. Chromosomes 1, 8, and 10 were most often affected. The results are compared with molecular genetic abnormalities recently reported in these lesions. This study strongly suggests that atypical endometrial hyperplasias are closely related to endometrioid carcinoma and should be considered precancerous lesions, contrary to simple hyperplasia, which is a benign disorder. The squamous component of one of the high-grade carcinomas showed genetic abnormalities similar to those of endometrioid carcinoma and therefore does not represent squamous metaplasia but is an integral part of the malignant process.
Asunto(s)
Aberraciones Cromosómicas , Hiperplasia Endometrial/genética , Neoplasias Endometriales/genética , Hibridación de Ácido Nucleico , Adulto , Anciano , Cromosomas Humanos Par 1 , Cromosomas Humanos Par 10 , Cromosomas Humanos Par 14 , Cromosomas Humanos Par 18 , Cromosomas Humanos Par 7 , Cromosomas Humanos Par 8 , Cromosomas Humanos Par 9 , Hiperplasia Endometrial/patología , Femenino , Humanos , Persona de Mediana EdadRESUMEN
Observation of genetic alterations that appear consistently in specific types and stages of cancer provides a strong impetus to cancer geneticists to focus their investigations on the exploration of such volatile regions of the human genome. Introduction of powerful molecular cytogenetic and molecular genetic methods in recent years permits more detailed analysis, which will help researchers in their efforts to determine if such areas of the human genome have a functional role in the initiation and progressive development of leukemias and solid tumors. This discussion will focus on several provocative molecular cytogenetic tools that are currently available to localize potential cancer-associated genes and on how these methods are being used in conjunction with the current modes of analysis, including cytogenetics and somatic cell genetics. In addition, we will explore how these methods will help to isolate and dissect recently discovered cancer-associated genes within the human genome. All of these methods used in combination with each other will provide essential DNA markers for future diagnostic and prognostic evaluation of cancer.
Asunto(s)
Mapeo Cromosómico , Neoplasias/genética , Animales , Aberraciones Cromosómicas , ADN de Neoplasias/aislamiento & purificación , Humanos , Hibridación de Ácido NucleicoRESUMEN
We report cytogenetic findings in a case of grade III chondrosarcoma. Complex clonal chromosome aberrations including monosomy of chromosomes 4, 8, 13, and a consistent t(5;14)(q23;p12) were observed in all cells. There were no structural or numerical anomalies involving chromosome 12. The complexity of the chromosome aberrations reflect the advanced stage of this chondrosarcoma; we suggest a possible involvement of the EXT1 gene located on chromosome 8.
Asunto(s)
Condrosarcoma/genética , Aberraciones Cromosómicas , Neoplasias Femorales/genética , Anciano , Condrosarcoma/patología , Femenino , Neoplasias Femorales/patología , Humanos , Cariotipificación , Monosomía , Recurrencia Local de Neoplasia , Translocación GenéticaRESUMEN
The integration sites of human papillomavirus (HPV) DNA within the cervical carcinoma cell line C4-I and a primary cervical tumor were mapped by in situ hybridization. Cloned cellular sequences flanking the integrated viral DNA were used as probes. For the cell line, the viral integration site was mapped to chromosome region 8q21-q22.3, while in the primary tumor chromosome band 3p21 was the target for integration. The HPV DNA integration appears to occur in the vicinity of fragile sites, oncogenes, and chromosome breakpoints that are characteristic of hematologic malignancies and solid tumors. The integration of HPV may thus promote chromosome changes in cancer cells.
Asunto(s)
Fragilidad Cromosómica , Mapeo Cromosómico , Oncogenes , Papillomaviridae/genética , Recombinación Genética , Neoplasias del Cuello Uterino/genética , Células Cultivadas , Bandeo Cromosómico , Sitios Frágiles del Cromosoma , ADN de Neoplasias/genética , ADN Viral/genética , Femenino , Marcadores Genéticos , Humanos , Cariotipificación , Masculino , Hibridación de Ácido Nucleico , Células Tumorales Cultivadas , Infecciones Tumorales por Virus/genética , Neoplasias del Cuello Uterino/microbiologíaRESUMEN
With a constant region probe (c-lambda) from the immunoglobulin light chain gene cluster, we performed in situ hybridization to bone marrow chromosome preparations from a patient with a Ph-positive form of acute lymphocytic leukemia. Results in this patient indicate that the immunoglobulin chain genes were involved in the 9;22 chromosome rearrangement.
Asunto(s)
Cromosomas Humanos 6-12 y X , Cadenas Ligeras de Inmunoglobulina/genética , Leucemia Linfoide/genética , Cromosoma Filadelfia , Mapeo Cromosómico , Genes , Humanos , Hibridación de Ácido NucleicoRESUMEN
This case presents a Caucasian girl diagnosed with early pre-B cell acute lymphoblastic leukemia at age 2 years. The only chromosomal anomaly detected in her bone marrow cells at this time was an add(12p). By age 4 years, she had a bone marrow and central nervous system (CNS) relapse of ALL and was treated with chemotherapy that included etoposide. She was in complete remission for 2 years following chemotherapy with etoposide, but later developed therapy-related acute myeloid leukemia (t-AML). At this time, a t(11;19)(q23;p13.3) rearrangement was detected in her bone marrow cells. The AML relapsed again 1 year after allogeneic bone marrow transplant (BMT). The presence of a chromosome 11 abnormality involving band 11q23 in this patient suggests that the transformation from ALL to t-AML was a consequence of etoposide included in her chemotherapy. Studies have shown that the 11q23 breakpoint in the t(11;19) rearrangement is consistent, and involves the MLL gene in t-AML patients. However, the breakpoint in 19p is variable in that it could be located either at 19p13.1 or 19p13.3 and thus could involve either of two genes: ELL (11-19 lysine-rich leukemia gene) on 19p13.1 or ENL (11-19 leukemia gene) on 19p13.3. In this study, the t(11;19)(q23;p13.3) was further characterized and the breakpoint regions were defined by fluorescence in situ hybridization (FISH) analysis.
Asunto(s)
Cromosomas Humanos Par 11 , Cromosomas Humanos Par 19 , Hibridación Fluorescente in Situ , Leucemia Mieloide Aguda/genética , Neoplasias Primarias Secundarias/genética , Proto-Oncogenes , Factores de Transcripción , Translocación Genética , Preescolar , Proteínas de Unión al ADN/genética , Femenino , N-Metiltransferasa de Histona-Lisina , Humanos , Proteína de la Leucemia Mieloide-LinfoideRESUMEN
We analyzed cytogenetic abnormalities in 10 cases of primary uveal melanoma. Clonal chromosomal abnormalities were present in nine cases. Chromosome 6 was most commonly affected (seven cases) and included gain of material from 6 and/or loss of material from 6q. Trisomy of chromosome 8 or gain in material from 8q, mostly in the form of an i(8q) resulting in three to five copies of the 8q segment was seen in six cases. Monosomy of chromosome 3 and rearrangements of chromosome 9 were less frequent and were altered in three cases each. Clinical, histopathologic, and cytogenetic abnormalities are correlated.
Asunto(s)
Aberraciones Cromosómicas , Melanoma/genética , Neoplasias de la Úvea/genética , Adulto , Anciano , Anciano de 80 o más Años , Cromosomas Humanos Par 6 , Cromosomas Humanos Par 8 , Femenino , Humanos , Masculino , Melanoma/patología , Persona de Mediana Edad , Neoplasias de la Úvea/patologíaRESUMEN
Nineteen cell lines derived from various malignant tissues of 15 patients with small cell carcinoma of the lung (SCCL) have been studied. The results showed heterogeneity in all cell lines, with no one consistent abnormality among them. Cell lines from 11 of the patients had minute and double minute chromosomes, and cell lines from 2 patients had abnormally banding regions, designated as ABRs, as distinguished from homogeneously staining regions (HSRs). The latter 2 and several of the former cell lines were derived from specimens taken before the patients were placed on therapy. All but 2 of the cell lines had a constant marker load, consisting of 24%-35% of the complement. Some markers remained stable through months and years of culture life, while other markers came and went. Chromosomes #1, #6 and #11 were most frequently involved in marker formation in the cell lines, and these were compared to similar markers in direct bone marrow preparations. Chromosome #1 markers were of variable structure, whereas #6 and #11 most often took the form of 6q- and 11p+ markers, with breakpoints most frequently at 6q23-25 and 11p11-12. A 3p- marker was found in a minority of cell lines. All of these markers were also found in direct marrow preparations from some patients with SCCL. Nonmonoclonal tumors arose from inoculation of bimodal cell lines into nude mice, but population selection by undetermined mechanism was evident. Cytogenetic parameters showed no positive correlation with hormone production by these cell lines.