RESUMO
Myeloproliferative neoplasms (MPN) are multiple disease entities characterized by clonal expansion of one or more of the myeloid lineages (i.e. granulocytic, erythroid, megakaryocytic and mast cell). JAK2 mutations, such as the common V617F substitution and the less common exon 12 mutations, are frequently detected in such tumor cells and have been incorporated into the diagnostic criteria published by the World Health Organization since 2008. However, the mechanism by which these mutations contribute to MPN development is poorly understood. We examined gene expression profiles of MPN patients focusing on genes in the JAK-STAT signaling pathway using low-density real-time PCR arrays. We identified the following 2 upregulated genes in MPN patients: a known target of the JAK-STAT axis, SOCS3, and a potentially novel target, SPI1, encoding PU.1. Induction of PU.1 expression by JAK2 V617F in JAK2-wildtype K562 cells and its downregulation by JAK2 siRNA transfection in JAK2 V617F-positive HEL cells supported this possibility. We also found that the ABL1 kinase inhibitor imatinib was very effective in suppressing PU.1 expression in BCR-ABL1-positive K562 cells but not in HEL cells. This suggests that PU.1 expression is regulated by both JAK2 and ABL1. The contribution of the two kinases in driving PU.1 expression was dominant for JAK2 and ABL1 in HEL and K562 cells, respectively. Therefore, PU.1 may be a common transcription factor upregulated in MPN. PU.1 is a transcription factor required for myeloid differentiation and is implicated in erythroid leukemia. Therefore, expression of PU.1 downstream of activated JAK2 may explain why JAK2 mutations are frequently observed in MPN patients.
Assuntos
Substituição de Aminoácidos/genética , Neoplasias da Medula Óssea/sangue , Neoplasias da Medula Óssea/genética , Janus Quinase 2/genética , Transtornos Mieloproliferativos/sangue , Proteínas Proto-Oncogênicas/genética , Transativadores/genética , Regulação para Cima/genética , Idoso , Neoplasias da Medula Óssea/enzimologia , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Janus Quinase 2/metabolismo , Masculino , Pessoa de Meia-Idade , Transtornos Mieloproliferativos/enzimologia , Transtornos Mieloproliferativos/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Transcrição STAT/genética , Fatores de Transcrição STAT/metabolismo , Transdução de Sinais/genética , Proteína 1 Supressora da Sinalização de Citocina , Proteína 3 Supressora da Sinalização de Citocinas , Proteínas Supressoras da Sinalização de Citocina/genética , Proteínas Supressoras da Sinalização de Citocina/metabolismoRESUMO
Feasibility of chromosomal manipulation in mammalian cells was first reported 15 years ago. Although this technique is useful for precise understanding of gene regulation in the chromosomal context, a limited number of laboratories have used it in actual practice because of associated technical difficulties. To overcome the practical hurdles, we developed a Cre-mediated chromosomal recombination system using fluorescent proteins and various site-specific recombinases. These techniques enabled quick construction of targeting vectors, easy identification of chromosome-rearranged cells, and rearrangement leaving minimum artificial elements at junctions. Applying this system to a human cell line, we successfully recapitulated two types of pathogenic chromosomal translocations in human diseases: MYC/IgH and BCR/ABL1. By inducing recombination between two loxP sites targeted into the same chromosome, we could mark cells harboring deletion or duplication of the inter-loxP segments with different colors of fluorescence. In addition, we demonstrated that the intrachromosomal recombination frequency is inversely proportional to the distance between two recombination sites, implicating a future application of this frequency as a proximity sensor. Our method of chromosomal manipulation can be employed for particular cell types in which gene targeting is possible (e.g. embryonic stem cells). Experimental use of this system would open up new horizons in genome biology, including the establishment of cellular and animal models of diseases caused by translocations and copy-number variations.
Assuntos
Cromossomos/ultraestrutura , DNA Nucleotidiltransferases/genética , Corantes Fluorescentes/metabolismo , Técnicas Genéticas , Vetores Genéticos , Animais , Linhagem Celular , Drosophila , Dosagem de Genes , Marcação de Genes , Humanos , Hibridização in Situ Fluorescente , Integrases/genética , Recombinação Genética , Translocação GenéticaRESUMO
Acute lymphoblastic leukemia (ALL) with chromosome aberration t(8;14)(q11.2;q32) mostly affects patients younger than 20 years old. One third of patients with this translocation have been reported to have Down syndrome. This translocation has been reported rarely in patients over the age of 50. Here we report a 71-year-old male ALL patient who carried t(8;14)(q11.2;q32). Fluorescence in situ hybridization (FISH) analysis revealed the involvement of CCAAT enhancer-binding protein delta (CEBPD) gene on chromosome 8, and IgH gene on chromosome 14. This case provides a new aspect for considering this clinical entity.
Assuntos
Cromossomos Humanos Par 14/genética , Cromossomos Humanos Par 8/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Translocação Genética , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Proteína delta de Ligação ao Facilitador CCAAT/genética , Aberrações Cromossômicas , Evolução Fatal , Genes de Cadeia Pesada de Imunoglobulina , Humanos , Hibridização in Situ Fluorescente , Masculino , Recidiva Local de Neoplasia/patologiaRESUMO
PURPOSE: Seventy to eighty percent of rhabdomyosarcoma (RMS) tumors retain wild-type p53. The tumor suppressor p53 plays a central role in inducing cell cycle arrest or apoptosis in response to various stresses. p53 protein levels are regulated by MDM2 through ubiquitin-dependent degradation. In this study, we evaluated whether nutlin-3, a recently developed small-molecule antagonist of MDM2, has an effect on p53-dependent cell cycle arrest and apoptosis in cultured human RMS cell lines. EXPERIMENTAL DESIGN: Five RMS cell lines with different p53 statuses and MDM2 expression levels were treated with nutlin-3. Gene expression patterns, cell viability, cell cycle, and apoptosis after nutlin-3 treatment, and antitumor activity of combination treatment with vincristine or actinomycin D were assessed. RESULTS: Significant p53 activation was observed in wild-type p53 cell lines after nutlin-3 treatment. p53 activation led to cell cycle arrest in parallel with increased p21 expression. Furthermore, these cell lines underwent p53-dependent apoptosis, concomitant with elevation of proapoptotic genes and activation of caspase-3. The effect of nutlin-3 was almost the same in terms of half maximal inhibitory concentration and apoptosis whether or not MDM2 was overexpressed. Nutlin-3 did not induce either cell cycle arrest or apoptosis in p53 mutant cell lines. A combination of vincristine or actinomycin D with nutlin-3 enhanced the antitumor activity in RMS cell lines with wild-type p53. CONCLUSIONS: Nutlin-3 effectively restored p53 function in both normal MDM2 expression and MDM2 overexpression RMS cell lines with wild-type p53. p53 restoration therapy is a potential therapeutic strategy for refractory RMS with wild-type p53.
Assuntos
Antibióticos Antineoplásicos/farmacologia , Apoptose , Ciclo Celular/efeitos dos fármacos , Imidazóis/farmacologia , Neoplasias de Tecido Muscular/metabolismo , Piperazinas/farmacologia , Proteínas Proto-Oncogênicas c-mdm2/antagonistas & inibidores , Rabdomiossarcoma/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Dactinomicina/farmacologia , Sinergismo Farmacológico , Humanos , Mutação de Sentido Incorreto/genética , Proteína Supressora de Tumor p53/agonistas , Proteína Supressora de Tumor p53/genética , Vincristina/farmacologiaRESUMO
Rhabdomyosarcoma is the most common soft tissue tumor seen in children and young adults, and it can be classified into 2 major histological subtypes, alveolar and embryonal. In the alveolar subtype, 2 recurrent chromosomal translocations, t(2;13)(q35;q14) and its variant t(1;13)(p36;q14), have been identified as the specific cytogenetic abnormalities. These translocations produce the PAX3-FOXO1 and PAX7-FOXO1 fusion genes, respectively. In the embryonal subtype, however, no recurrent chromosomal abnormalities have been identified. In this study, we analyzed the complex chromosomal translocation in one case with embryonal rhabdomyosarcoma by means of spectral karyotyping (SKY) and identified a novel translocation involving chromosome band 2q35, which is the locus of PAX3 gene. Furthermore, we identified the novel PAX3 rearrangement using fluorescence in situ hybridization (FISH) analysis. Additional identification of the partner gene may help disclose the molecular mechanism of the development of this embryonal subtype.
Assuntos
Mutação , Fatores de Transcrição Box Pareados/genética , Rabdomiossarcoma Embrionário/genética , Neoplasias Urogenitais/genética , Análise Citogenética , Análise Mutacional de DNA , Humanos , Lactente , Masculino , Proteínas de Fusão Oncogênica/genética , Fator de Transcrição PAX3 , Proteínas Recombinantes de Fusão/análise , Proteínas Recombinantes de Fusão/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
A 17-year-old boy developed therapy-related acute myeloid leukemia (t-AML) 3 years after the cessation of chemo- and radiotherapy for undifferentiated sarcoma of the liver. At the onset of the t-AML, his white blood cell count was 900/microL with a 46,XY,t(2;3)(p21;q26),del(5)(q?) karyotype. Despite intensive chemotherapy and two hematopoietic stem cell transplants, he died of the leukemia. At the terminal phase, his white blood cell count surpassed 30,000/microL and the Philadelphia (Ph) chromosome appeared. Expression of EVI1 in bone marrow cells was remarkably high at the onset of t-AML, although it was not detected at the end of therapy for the sarcoma. Polymerase chain reaction analysis of bone marrow cells revealed that mRNA for the bcr-abl chimera was negative at the onset of t-AML and positive at the terminal phase. These results suggest that EVI1 overexpression was the major factor contributing to leukemogenesis, and the late appearance of the Ph chromosome is closely associated with the progression to an aggressive form of leukemia.
Assuntos
Proteínas de Ligação a DNA/genética , Segunda Neoplasia Primária/genética , Cromossomo Filadélfia , Proto-Oncogenes/genética , Fatores de Transcrição/genética , Adolescente , Proteínas de Fusão bcr-abl/genética , Proteínas de Fusão bcr-abl/metabolismo , Regulação Leucêmica da Expressão Gênica , Humanos , Hibridização in Situ Fluorescente , Leucemia Mieloide Aguda/etiologia , Leucemia Mieloide Aguda/genética , Neoplasias Hepáticas/terapia , Proteína do Locus do Complexo MDS1 e EVI1 , Masculino , RNA Mensageiro/metabolismo , Sarcoma/terapia , Regulação para CimaRESUMO
A new cell line, designated UCH1, was established from a patient with splenic marginal zone lymphoma (SMZL). UCH1 cells feature a mature B-cell phenotype, characterized by surface IgM +, kappa+, CD5-, CD10-, CD19+ and CD20+. The BCL2 and BCL6 genes retained their germ-line configurations and overexpression of cyclin D1 was not detected. UCH1 cells carry numerical and structural aberrations in chromosome 3, but these were too complex to be analyzed with the conventional G-banding method. Spectral karyotyping (SKY) and fluorescence in situ hybridization analysis clearly demonstrated the presence of a balanced translocation between chromosomes 8 and 14 [t(8;14)(q24;q32)] in the complex aberrations involving chromosome 3. The results of Southern blot analysis supported this finding by showing rearrangement of the c-myc gene in UCH1 cells. SKY analysis also identified a translocation involving chromosome band 18q21, to which BCL2 and MALT1 genes were assigned, suggesting their implication in the development or progression of SMZL.
Assuntos
Linhagem Celular Tumoral , Cromossomos Humanos Par 3 , Linfoma/genética , Linfoma/patologia , Translocação Genética , Idoso , Linfócitos B/metabolismo , Rearranjo Gênico , Humanos , Imunoglobulina M/metabolismo , Imunofenotipagem , Hibridização in Situ Fluorescente , Cariotipagem , Leucócitos Mononucleares/metabolismo , Masculino , Proteínas Proto-Oncogênicas c-myc/metabolismoRESUMO
A new cell line, STR-428 was established from ascites tumor cells of a malignant effusion lymphoma patient without human herpes virus-8 (HHV-8) infection. STR-428 cells showed an immunophenotype of mature B-cells and produced few cytokines related to lymphomatous effusion. Karyotypic and genetic analysis revealed complex translocations including t(14;18)(q32;q21) effecting IgH/BCL2 and der(8)t(3;8)(q27;q24) involving c-MYC. STR-428 represents a unique, B-cell lymphoma cell line carrying concurrent rearrangement of BCL2 and c-MYC genes with features distinct from those of HHV-8-related primary effusion lymphoma. This cell line may be a valuable tool, other than HHV-8, to investigate the pathogenesis of primary lymphomatous effusion.
Assuntos
Herpesvirus Humano 8/fisiologia , Linfoma/genética , Derrame Pleural Maligno/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-myc/genética , Translocação Genética , Southern Blotting , Cromossomos Humanos Par 14/genética , Cromossomos Humanos Par 18/genética , Citocinas , Humanos , Técnicas Imunoenzimáticas , Imunofenotipagem , Hibridização in Situ Fluorescente , Linfoma/metabolismo , Linfoma/virologia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Células Tumorais CultivadasRESUMO
We conducted a retrospective analysis of the clinical features of 20 patients with severe eosinophilia at our institution, including 10 cases of hypereosinophilic syndrome (HES) (5 definite and 5 probable cases) and 10 cases of other eosinophilic disorders. Of the 20 patients, 14 initially received prednisolone treatment, which resulted in rapid improvement and normalization of eosinophilia within 8 weeks; however, 2 patients with splenomegaly showed poor control of eosinophilia in response to corticosteroid treatment. In addition, the FIP1L1-PDGFRA fusion gene was detected only in these 2 cases. One of the FIP1L1-PDGFRA - positive HES cases featured bone marrow fibrosis. Treatment of this patient with imatinib mesylate resulted in a dramatic improvement of eosinophilia, organomegaly, and the bone marrow fibrosis. Taken together, our data and previous reports suggest that FIP1L1-PDGFRA - positive HES is a distinct clinical entity with myeloproliferative features and showing a poor response to corticosteroid treatment.
Assuntos
Corticosteroides/uso terapêutico , Síndrome Hipereosinofílica/genética , Síndrome Hipereosinofílica/patologia , Transtornos Mieloproliferativos/genética , Proteínas de Fusão Oncogênica/genética , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/genética , Fatores de Poliadenilação e Clivagem de mRNA/genética , Adulto , Idoso , Medula Óssea/patologia , Eosinofilia/tratamento farmacológico , Eosinofilia/genética , Eosinofilia/patologia , Feminino , Fibrose/etiologia , Humanos , Síndrome Hipereosinofílica/tratamento farmacológico , Hibridização in Situ Fluorescente , Masculino , Pessoa de Meia-Idade , Prednisolona/uso terapêutico , Prognóstico , Estudos Retrospectivos , EsplenomegaliaRESUMO
Activation-induced cytidine deaminase (AID), which is essential to both class switch recombination and somatic hypermutation of the Ig gene, is expressed in many types of human B cell lymphoma/leukemia. AID is a potent mutator because it is involved in DNA breakage not only of Ig but also of other genes, including proto-oncogenes. Recent studies suggest that AID is required for chromosomal translocation involving cmyc and Ig loci. However, it is unclear whether AID plays other roles in tumorigenesis. We examined the effect of AID deficiency on the generation of surface Ig-positive B cell lymphomas in Emu-cmyc transgenic mice. Almost all lymphomas that developed in AID-deficient transgenic mice were pre-B cell lymphomas, whereas control transgenic mice had predominantly B cell lymphomas, indicating that AID is required for development of B but not pre-B cell lymphomas from cmyc overexpressing tumor progenitors. Thus, AID may play multiple roles in B cell lymphomagenesis.
Assuntos
Linfócitos B/citologia , Citidina Desaminase/fisiologia , Regulação Neoplásica da Expressão Gênica , Linfoma de Células B/metabolismo , Animais , Linfócitos B/enzimologia , Separação Celular , Dano ao DNA , Linfoma de Células B/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Modelos Biológicos , Mutação , Fenótipo , Proteínas Proto-Oncogênicas c-myc/metabolismo , Proteínas Proto-Oncogênicas c-pim-1/genética , Translocação GenéticaRESUMO
Some immortal cells use the alternative lengthening of telomeres (ALT) pathway to maintain their telomeres instead of telomerase. Previous studies revealed that homologous recombination (HR) contributes to the ALT pathway. To further elucidate molecular mechanisms, we inactivated Rad54 involved in HR, in mouse ALT embryonic stem (ES) cells. Although Rad54-deficient ALT ES cells showed radiosensitivity in line with expectation, cell growth and telomeres were maintained for more than 200 cell divisions. Furthermore, although MMC-stimulated sister chromatid exchange (SCE) was suppressed in the Rad54-deficient ALT ES cells, ALT-associated telomere SCE was not affected. This is the first genetic evidence that mouse Rad54 is dispensable for the ALT pathway.
Assuntos
Proteínas Nucleares/metabolismo , Telômero/metabolismo , Animais , Linhagem Celular , DNA Helicases , Células-Tronco Embrionárias/metabolismo , Camundongos , Camundongos Knockout , Proteínas Nucleares/deficiência , Proteínas Nucleares/genética , Recombinação Genética , Transdução de Sinais , Troca de Cromátide IrmãRESUMO
Nuclei isolated from green fluorescent protein-marked neurons in the cerebral cortex of juvenile mice (14-21 d after birth) were injected into enucleated oocytes that were allowed to develop into blastocysts. Embryonic stem (ES) cell lines were established from the inner cell mass of 76 cloned blastocysts after injecting 2026 neuronal nuclei. Some ES cells were injected individually into enucleated oocytes (nuclear transfer). Other ES cells were transferred into the blastocoeles of tetraploid blastocysts (tetraploid complementation). Two-cell embryos after nuclear transfer were transferred to the oviducts of surrogate mothers. Four (1.5%) of 272 nuclear-transferred two-cell embryos developed to term, and two (0.7%) developed into fertile adults. Nineteen (1.9%) of 992 tetraploid blastocysts receiving ES cells reached term, and 10 (1.0%) developed into adults. These findings demonstrate that some of the nuclei of differentiated neurons in the cerebral cortex of juvenile mice maintain developmental pluripotency.
Assuntos
Núcleo Celular/fisiologia , Núcleo Celular/ultraestrutura , Córtex Cerebral/fisiologia , Neurônios/citologia , Neurônios/fisiologia , Animais , Blastocisto/fisiologia , Córtex Cerebral/embriologia , Clonagem de Organismos , Embrião de Mamíferos , Marcadores Genéticos , Vetores Genéticos , Proteínas de Fluorescência Verde/análise , Proteínas de Fluorescência Verde/genética , Camundongos , Plasmídeos , Poliploidia , Células-Tronco/citologia , Células-Tronco/fisiologiaRESUMO
A 26-year-old man with idiopathic hypereosinophilic syndrome (HES) was treated with imatinib mesylate following a 5-year history of prednisolone therapy. The patient had hypereosinophilia (absolute eosinophil counts >1500/microL) occurring in cyclic oscillations as well as histologically diagnosed eosinophilic vasculitis, bursitis, and periodic soft-tissue swellings. Laboratory data revealed high levels of serum tryptase and increased numbers of mast cells in the bone marrow, but serum interleukin 5 levels were within the normal range. The disease initially responded well to 100 mg/day of imatinib mesylate but recurred 8 weeks later. Thereafter, a daily 200-mg dose was temporarily effective. Despite the response to imatinib, the FIP1L1-PDGFRA fusion gene was not detected by fluorescence in situ hybridization analysis. Additional molecular and cytogenetic studies showed neither translocations of platelet-derived growth factor receptor (PDGFR) genes nor mutations in the c-KIT or the PDGFR genes. Although imatinib mesylate is a choice of treatment for patients with HES, its precise molecular mechanism in individual cases remains to be clarified.
Assuntos
Antineoplásicos/farmacologia , Síndrome Hipereosinofílica/tratamento farmacológico , Piperazinas/farmacologia , Pirimidinas/farmacologia , Adulto , Benzamidas , Análise Mutacional de DNA , Humanos , Mesilato de Imatinib , Masculino , Receptores do Fator de Crescimento Derivado de Plaquetas/genética , Recidiva , Resultado do TratamentoRESUMO
A 2-year-old girl with Down syndrome (DS) developed acute megakaryoblastic leukemia (AMKL) following a transient myeloproliferative disorder (TMD). The blast cells showed an altered karyotype of 47,XX,r(7),+21c. Serial cytogenetic studies during the course of the illness showed rapid stepwise clonal chromosome changes, including a ring chromosome 7, associated with treatment refractoriness. We reviewed 10 published cases of Down syndrome-related AMKL (DS-AMKL) showing chromosome 7 abnormalities and found that these changes do not carry the same prognostic weight as for non-DS children. For DS-AMKL, therefore, other prognostic factors besides clonal cytogenetic changes need to be identified for planning optimal therapy.
Assuntos
Aberrações Cromossômicas , Transtornos Cromossômicos , Cromossomos Humanos Par 7 , Síndrome de Down/genética , Leucemia Megacarioblástica Aguda/genética , Pré-Escolar , Progressão da Doença , Síndrome de Down/complicações , Feminino , Humanos , Leucemia Megacarioblástica Aguda/complicações , Cromossomos em Anel , Cariotipagem Espectral , TrissomiaRESUMO
Diverse protocadherin-alpha genes (Pcdha, also called cadherin-related neuronal receptor or CNR) are expressed in the vertebrate brain. Their genomic organization involves multiple variable exons and a set of constant exons, similar to the immunoglobulin (Ig) and T-cell receptor (TCR) genes. This diversity can be used to distinguish neurons. Using polymorphisms that distinguish the C57BL/6 and MSM mouse strains, we analyzed the allelic expression of the Pcdha gene cluster in individual neurons. Single-cell analysis of Purkinje cells using multiple RT-PCR reactions showed the monoallelic and combinatorial expression of each variable exon in the Pcdha genes. This report is the first description to our knowledge of the allelic expression of a diversified receptor family in the central nervous system. The allelic and combinatorial expression of distinct variable exons of the Pcdha genes is a potential mechanism for specifying neuron identity in the brain.
Assuntos
Caderinas/genética , Variação Genética , Neurônios/metabolismo , Animais , Éxons , Expressão Gênica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Modelos Genéticos , Dados de Sequência Molecular , Família Multigênica , Células de Purkinje/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Activation-induced cytidine deaminase (AID) is required for the DNA cleavage step in immunoglobulin class switch recombination (CSR). AID is proposed to deaminate cytosine to generate uracil (U) in either mRNA or DNA. In the second instance, DNA cleavage depends on uracil DNA glycosylase (UNG) for removal of U. Using phosphorylated histone gamma-H2AX focus formation as a marker of DNA cleavage, we found that the UNG inhibitor Ugi did not inhibit DNA cleavage in immunoglobulin heavy chain (IgH) locus during CSR, even though Ugi blocked UNG binding to DNA and strongly inhibited CSR. Strikingly, UNG mutants that had lost the capability of removing U rescued CSR in UNG-/- B cells. These results indicate that UNG is involved in the repair step of CSR yet by an unknown mechanism. The dispensability of U removal in the DNA cleavage step of CSR requires a reconsideration of the model of DNA deamination by AID.
Assuntos
Linfócitos B/fisiologia , DNA Glicosilases/metabolismo , DNA/metabolismo , Genes de Imunoglobulinas , Switching de Imunoglobulina , Animais , Linfócitos B/enzimologia , Linfócitos B/imunologia , Linhagem Celular Tumoral , Citidina Desaminase/metabolismo , DNA Glicosilases/antagonistas & inibidores , DNA Glicosilases/genética , Reparo do DNA , Cadeias Pesadas de Imunoglobulinas/genética , Região de Troca de Imunoglobulinas , Camundongos , Mutação , Testes de Precipitina , Recombinação Genética , Transfecção , Uracila/metabolismo , Uracila-DNA Glicosidase , Proteínas Virais/metabolismoRESUMO
Acute myeloblastic leukemia cases carrying the translocation t(8;16) (p11;p13) are characterized by the M4 and M5 subtypes, erythrophagocytosis by the blast cells and a poor prognosis, suggesting a new clinical entity. The t(8;16) fuses the MOZ gene which encodes a histone acetyltransferase, located on 8p11 with the CBP gene which also encodes a histone acetyltransferase, located on 16p13, and recent reports suggested that the chimeric transcription MOZ-CBP is essential for leukemogenesis. A 68-year-old woman who had been treated mainly with paclitaxel and carboplatin for preceding ovarian cancer was admitted to our hospital, complaining of right breast mass. She was diagnosed as having breast cancer and acute monocytic leukemia (M5b). Cytogenetic study with spectral karyotyping analysis revealed the development of 47 XX, + 8, t(8;16)(p11;p13). Eleven cases of therapy-related t(8;16) leukemia including the present case have been reported, but prior treatment with paclitaxel and carboplatin-based chemotherapy has never been reported. The relation of histone acetylase and therapy-related leukemia is discussed.
Assuntos
Cromossomos Humanos Par 16 , Cromossomos Humanos Par 8 , Leucemia Monocítica Aguda/genética , Segunda Neoplasia Primária/genética , Translocação Genética , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Carboplatina/efeitos adversos , Transformação Celular Neoplásica/genética , Feminino , Humanos , Leucemia Monocítica Aguda/induzido quimicamente , Segunda Neoplasia Primária/induzido quimicamente , Neoplasias Ovarianas/complicações , Neoplasias Ovarianas/tratamento farmacológico , Paclitaxel/efeitos adversosRESUMO
We have developed spectral color banding (SCAN) as a new chromosome banding technique based on spectral analysis of differentially labeled chromosome band-specific painting probes. In this study, we succeeded in displaying a multicolor banding pattern for chromosome 3, which was almost identical to the pattern obtained with the corresponding G-banding. We applied this method to metaphase cells from different normal male donors with various levels of G-banding resolution, ranging from 250 to 850 bands per haploid set. The same multicolor banding pattern was observed in all samples regardless of the length of the chromosomes or the quality of the G-banding. We then used SCAN in a diffuse large B-cell lymphoma case for a complete analysis of the intrachromosomal change for chromosome 3, which could not be fully characterized by G-banding or even by spectral karyotyping (SKY). SCAN could detect the duplicated segment and identify the origin of the chromosome band on the basis of the specific spectral color of each band. This study demonstrates that SCAN is a useful tool for full characterization of chromosomal abnormalities not identified by SKY.