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1.
Cancer Sci ; 112(5): 1888-1898, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33560542

RESUMEN

Circular RNAs (circRNAs) have been identified to exert vital biological functions and can be used as new biomarkers in a number of tumors. However, little is known about the functions of circRNAs in myelodysplastic syndrome (MDS). Here, we aimed to investigate circRNA expression profiles and to investigate the functional and clinical value of circRNAs in MDS. Differential expression of circRNAs between MDS and control subjects was analyzed using circRNA arrays, in which we identified 145 upregulated circRNAs and 224 downregulated circRNAs. Validated by real-time quantitative PCR between 100 MDS patients and 20 controls, three upregulated (hsa_circRNA_100352, hsa_circRNA_104056, and hsa_circRNA_104634) and three downregulated (hsa_circRNA_103846, hsa_circRNA_102817, and hsa_circRNA_102526) circRNAs matched the arrays. The receiver operating characteristic curve analysis of these circRNAs showed that the area under the curve was 0.7266, 0.8676, 0.7349, 0.7091, 0.8806, and 0.7472, respectively. Kaplan-Meier survival analysis showed that only hsa_circRNA_100352, hsa_circRNA_104056, and hsa_circRNA_102817 were significantly associated with overall survival. Furthermore, we generated a competing endogenous RNA network focused on hsa_circRNA_100352, hsa_circRNA_104056, and hsa_circRNA_102817. Analyses using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes showed that the three circRNAs were linked with some important cancer-related functions and pathways.


Asunto(s)
Biomarcadores de Tumor/metabolismo , Síndromes Mielodisplásicos/metabolismo , ARN Circular/metabolismo , Anciano , Anemia Refractaria/genética , Anemia Refractaria/metabolismo , Anemia Refractaria con Exceso de Blastos/genética , Anemia Refractaria con Exceso de Blastos/metabolismo , Anemia Sideroblástica/genética , Anemia Sideroblástica/metabolismo , Área Bajo la Curva , Biomarcadores de Tumor/genética , Médula Ósea/metabolismo , Estudios de Casos y Controles , Regulación hacia Abajo , Femenino , Humanos , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/sangre , Síndromes Mielodisplásicos/genética , Síndromes Mielodisplásicos/mortalidad , ARN Circular/genética , Curva ROC , Reacción en Cadena en Tiempo Real de la Polimerasa , Estadísticas no Paramétricas , Regulación hacia Arriba
2.
Blood ; 135(25): 2271-2285, 2020 06 18.
Artículo en Inglés | MEDLINE | ID: mdl-32202636

RESUMEN

SETD2, the histone H3 lysine 36 methyltransferase, previously identified by us, plays an important role in the pathogenesis of hematologic malignancies, but its role in myelodysplastic syndromes (MDSs) has been unclear. In this study, low expression of SETD2 correlated with shortened survival in patients with MDS, and the SETD2 levels in CD34+ bone marrow cells of those patients were increased by decitabine. We knocked out Setd2 in NUP98-HOXD13 (NHD13) transgenic mice, which phenocopies human MDS, and found that loss of Setd2 accelerated the transformation of MDS into acute myeloid leukemia (AML). Loss of Setd2 enhanced the ability of NHD13+ hematopoietic stem and progenitor cells (HSPCs) to self-renew, with increased symmetric self-renewal division and decreased differentiation and cell death. The growth of MDS-associated leukemia cells was inhibited though increasing the H3K36me3 level by using epigenetic modifying drugs. Furthermore, Setd2 deficiency upregulated hematopoietic stem cell signaling and downregulated myeloid differentiation pathways in the NHD13+ HSPCs. Our RNA-seq and chromatin immunoprecipitation-seq analysis indicated that S100a9, the S100 calcium-binding protein, is a target gene of Setd2 and that the addition of recombinant S100a9 weakens the effect of Setd2 deficiency in the NHD13+ HSPCs. In contrast, downregulation of S100a9 leads to decreases of its downstream targets, including Ikba and Jnk, which influence the self-renewal and differentiation of HSPCs. Therefore, our results demonstrated that SETD2 deficiency predicts poor prognosis in MDS and promotes the transformation of MDS into AML, which provides a potential therapeutic target for MDS-associated acute leukemia.


Asunto(s)
Anemia Refractaria con Exceso de Blastos/patología , Calgranulina B/fisiología , N-Metiltransferasa de Histona-Lisina/deficiencia , N-Metiltransferasa de Histona-Lisina/fisiología , Leucemia Mieloide Aguda/etiología , Anemia Refractaria con Exceso de Blastos/genética , Anemia Refractaria con Exceso de Blastos/metabolismo , Animales , Calgranulina B/biosíntesis , Calgranulina B/genética , Transformación Celular Neoplásica , Células Cultivadas , Decitabina/farmacología , Regulación hacia Abajo , Regulación Leucémica de la Expresión Génica , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/patología , Código de Histonas/efectos de los fármacos , N-Metiltransferasa de Histona-Lisina/biosíntesis , N-Metiltransferasa de Histona-Lisina/genética , Proteínas de Homeodominio/genética , Humanos , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/mortalidad , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Síndromes Mielodisplásicos/patología , Proteínas de Complejo Poro Nuclear/genética , Proteínas de Fusión Oncogénica/genética , Pronóstico , Proteínas Recombinantes/uso terapéutico , Factores de Tiempo , Análisis de Matrices Tisulares , Transcriptoma
3.
Leukemia ; 29(5): 1092-103, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25428262

RESUMEN

The splicing factor SF3B1 is the most commonly mutated gene in the myelodysplastic syndrome (MDS), particularly in patients with refractory anemia with ring sideroblasts (RARS). We investigated the functional effects of SF3B1 disruption in myeloid cell lines: SF3B1 knockdown resulted in growth inhibition, cell cycle arrest and impaired erythroid differentiation and deregulation of many genes and pathways, including cell cycle regulation and RNA processing. MDS is a disorder of the hematopoietic stem cell and we thus studied the transcriptome of CD34(+) cells from MDS patients with SF3B1 mutations using RNA sequencing. Genes significantly differentially expressed at the transcript and/or exon level in SF3B1 mutant compared with wild-type cases include genes that are involved in MDS pathogenesis (ASXL1 and CBL), iron homeostasis and mitochondrial metabolism (ALAS2, ABCB7 and SLC25A37) and RNA splicing/processing (PRPF8 and HNRNPD). Many genes regulated by a DNA damage-induced BRCA1-BCLAF1-SF3B1 protein complex showed differential expression/splicing in SF3B1 mutant cases. This is the first study to determine the target genes of SF3B1 mutation in MDS CD34(+) cells. Our data indicate that SF3B1 has a critical role in MDS by affecting the expression and splicing of genes involved in specific cellular processes/pathways, many of which are relevant to the known RARS pathophysiology, suggesting a causal link.


Asunto(s)
Regulación Neoplásica de la Expresión Génica , Células Madre Hematopoyéticas/citología , Síndromes Mielodisplásicos/genética , Fosfoproteínas/genética , Ribonucleoproteína Nuclear Pequeña U2/genética , Células Madre/citología , Empalme Alternativo , Anemia Refractaria con Exceso de Blastos/genética , Anemia Refractaria con Exceso de Blastos/metabolismo , Antígenos CD34/metabolismo , Ciclo Celular , Diferenciación Celular , Proliferación Celular , Exones , Femenino , Perfilación de la Expresión Génica , Técnicas de Silenciamiento del Gen , Heterocigoto , Homeostasis , Humanos , Células K562 , Masculino , Mutación , Síndromes Mielodisplásicos/metabolismo , Fosfoproteínas/metabolismo , Mutación Puntual , ARN/genética , Empalme del ARN , Factores de Empalme de ARN , Ribonucleoproteína Nuclear Pequeña U2/metabolismo , Análisis de Secuencia de ARN
4.
Am J Hematol ; 89(8): E109-15, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24723457

RESUMEN

The SF3B1 mutation can be detected in patients with myelodysplastic syndrome (MDS), but the report regarding the association of this mutation with other genetic alterations and its stability during disease progression is limited. In this study, SF3B1 mutations were identified in 10% of total cohort of 479 MDS patients and 61.8% of 34 patients with refractory anemia with ring sideroblasts (RARS). SF3B1 mutations were closely associated with older age, higher platelet counts, lower lactate dehydrogenase levels, good-risk cytogenetics, and mutations of DNMT3A, but inversely related to ASXL1 mutations. Most SF3B1-mutated patients had concurrent other genetic alterations, including DNMT3A and RUNX1 mutations. There was no prognostic difference between patients with SF3B1 mutations and those without. Sequential studies in 417 samples from 142 patients demonstrated that all SF3B1-mutated patients retained the same mutations during disease evolution with the exception of two patients who lost the mutation after allogeneic hematopoietic stem cell transplantation, whereas none of the SF3B1-wild patients acquired a novel mutation during clinical follow-ups. In conclusion, the patients with SF3B1 mutations had distinct clinic-biologic features. SF3B1 mutations, accompanied with other genetic alterations, especially DNMT3A mutations, may play a role in the development of MDS, but have little role in disease progression.


Asunto(s)
Anemia Refractaria con Exceso de Blastos/genética , ADN (Citosina-5-)-Metiltransferasas/genética , Tasa de Mutación , Síndromes Mielodisplásicos/genética , Fosfoproteínas/genética , Ribonucleoproteína Nuclear Pequeña U2/genética , Adolescente , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Anemia Refractaria con Exceso de Blastos/metabolismo , Anemia Refractaria con Exceso de Blastos/patología , Plaquetas/metabolismo , Plaquetas/patología , Subunidad alfa 2 del Factor de Unión al Sitio Principal/genética , Subunidad alfa 2 del Factor de Unión al Sitio Principal/metabolismo , ADN (Citosina-5-)-Metiltransferasas/metabolismo , ADN Metiltransferasa 3A , Progresión de la Enfermedad , Femenino , Humanos , L-Lactato Deshidrogenasa/genética , L-Lactato Deshidrogenasa/metabolismo , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/metabolismo , Síndromes Mielodisplásicos/patología , Fosfoproteínas/metabolismo , Factores de Empalme de ARN , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Ribonucleoproteína Nuclear Pequeña U2/metabolismo
5.
Cytometry B Clin Cytom ; 86(6): 383-96, 2014 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-24106013

RESUMEN

BACKGROUND: Single Cell Network Profiling (SCNP) is a multiparametric flow cytometry-based assay that quantifiably and simultaneously measures changes in intracellular signaling proteins in response to in vitro extracellular modulators at the single cell level. Myelodysplastic syndrome (MDS) is a heterogeneous clonal disorder of hematopoietic stem cells that occurs in elderly subjects and is characterized by dysplasia and ineffective hematopoiesis. The functional responsiveness of MDS bone marrow (BM) hematopoietic cells, including functionally distinct myeloid and erythroid precursor subsets, to hematopoietic growth factors (HGF) and the relationship of modulated signaling to disease characteristics is poorly understood. METHODS: SCNP was used first to examine the effects of age on erythropoietin (EPO) and granulocyte colony stimulating factor (GCSF)-induced signaling in myeloid, nucleated red blood cells (nRBC), and CD34 expressing cell subsets in healthy BM (n = 15). SCNP was then used to map functional signaling profiles in low risk (LR) MDS (n = 7) for comparison to signaling in samples from healthy donors and to probe signaling associations within clinically defined subgroups. RESULTS: In healthy BM samples, signaling responses to HGF were quite homogeneous (i.e., tightly regulated) with age-dependent effects observed in response to EPO but not to GCSF. Despite the relatively small number of samples assayed in the study, LR MDS could be classified into distinct subgroups based on both cell subset frequency and signaling profiles. CONCLUSIONS: As a correlate of underlying genetic abnormalities, signal transduction analyses may provide a functional and potentially clinically relevant classification of MDS. Further evaluation in a larger cohort is warranted.


Asunto(s)
Células de la Médula Ósea/metabolismo , Péptidos y Proteínas de Señalización Intercelular/farmacología , Síndromes Mielodisplásicos/metabolismo , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Anemia Refractaria con Exceso de Blastos/metabolismo , Anemia Refractaria con Exceso de Blastos/patología , Células de la Médula Ósea/patología , Células Eritroides/metabolismo , Células Eritroides/patología , Eritropoyetina/farmacología , Citometría de Flujo/métodos , Factor Estimulante de Colonias de Granulocitos/metabolismo , Factor Estimulante de Colonias de Granulocitos/farmacología , Células Madre Hematopoyéticas/metabolismo , Células Madre Hematopoyéticas/patología , Humanos , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Persona de Mediana Edad , Síndromes Mielodisplásicos/patología , Células Mieloides/metabolismo , Células Mieloides/patología , Transducción de Señal , Adulto Joven
6.
DNA Cell Biol ; 31(10): 1563-70, 2012 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22873198

RESUMEN

Myelodysplastic syndrome (MDS) is a heterogeneous group of clonal hematopoietic stem cell diseases. Approximately 30% of patients with MDS will develop acute myeloid leukemia (AML). Immune dysregulation may contribute to MDS initiation and progression. The altered expression and clonal expansion of the Vß repertoire were observed in patients with MDS. To further examine the characteristic features of γδ(+)T cells in MDS, we investigated the distribution pattern and clonal expansion capacity of the T-cell receptor (TCR) Vγ and Vδ repertoire in patients with refractory anemia with excess of blasts (RAEB) and compared the difference between groups of patients with RAEB and RAEB-AML. Thirty-one patients with newly diagnosed MDS-RAEB were enrolled, and 9 of the 31 patients with RAEB developed AML (RAEB-AML). The TCR Vγ subfamily expression frequencies were similar in the RAEB and RAEB-AML patient groups. The number of the TCR Vδ subfamilies expressed in the RAEB group was higher than that in the RAEB-AML group. In most cases, a significantly higher Vδ4 subfamily expression frequency (63.64%, 14/22) could be detected in the RAEB group, whereas only 11.11% (1/9) was found in the RAEB-AML group (p=0.0079). At least one clonally expanded TCR Vδ subfamily member was detected in all cases in both groups. Vδ3 was the most frequent clonally expanded T cell subfamily member found in the RAEB and RAEB-AML group, while the most frequent clonally expanded T cell subfamily member in the RAEB-AML group was Vδ8 (87.5%, 7/8), which was significantly higher than that in the RAEB group (42.86%, 9/21; p=0.0307). In conclusion, the TCR Vδ subfamily expression pattern exhibited a marked restriction in patients with RAEB-AML. The lower Vδ4 frequency and higher clonally expanded Vδ8 T cell alterations were the characteristic features found in RAEB-AML. These results provide new data regarding the immunodeficiency and immune reactive characteristics of patients with RAEB and RAEB-AML.


Asunto(s)
Anemia Refractaria con Exceso de Blastos/metabolismo , Leucemia Mieloide Aguda/metabolismo , Síndromes Mielodisplásicos/metabolismo , Receptores de Antígenos de Linfocitos T gamma-delta/biosíntesis , Linfocitos T/metabolismo , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Progresión de la Enfermedad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven
7.
J Clin Exp Hematop ; 52(1): 63-6, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22706533

RESUMEN

A retrospective analysis of 71 patients newly diagnosed with refractory cytopenia with unilineage dysplasia (RCUD) revealed that 12 developed refractory anemia with an excess of blasts or acute myeloblastic leukemia. Before the diagnosis of RCUD was made, phenotypes of cells in the bone marrow (BM) blast region were analyzed using flow cytometry. Patients with RCUD were divided into two groups ; those with no progression (Group A) and those with disease progression later on (Group B). The cell composition in the BM blast region differed significantly between the groups : Group A showed higher percentages of B lymphoid cells but lower percentages of myeloid cells. A cut-off value of 20 for the CD33/CD10 ratio in the BM blast region clearly separated Group A from Group B. These results suggest that cell composition in the BM blast region evaluated by flow cytometry may indicate the progression of RCUD.


Asunto(s)
Anemia Refractaria con Exceso de Blastos/patología , Crisis Blástica/patología , Leucemia Mieloide Aguda/patología , Células Progenitoras Mieloides/patología , Células Precursoras de Linfocitos B/patología , Adulto , Anciano , Anciano de 80 o más Años , Anemia Refractaria con Exceso de Blastos/metabolismo , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Crisis Blástica/metabolismo , Progresión de la Enfermedad , Femenino , Estudios de Seguimiento , Humanos , Leucemia Mieloide Aguda/metabolismo , Masculino , Persona de Mediana Edad , Células Progenitoras Mieloides/metabolismo , Neprilisina/metabolismo , Células Precursoras de Linfocitos B/metabolismo , Lectina 3 Similar a Ig de Unión al Ácido Siálico
8.
Adv Ther ; 28 Suppl 2: 10-5, 2011 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21431505

RESUMEN

The management of myelodysplastic syndromes (MDS) in elderly patients is a significant clinical problem. The therapeutic options range from observation alone for patients with low-risk disease, lenalidomide for patients with 5q-syndrome, to 5-azacytidine (5-AZA) for patients with higher risk of disease. In this paper, we summarize the clinical course of three patients with high-risk MDS treated with 5-AZA as well as the management and supportive care measures for adverse events. As expected, based on available clinical trials data, the agent resulted in clinical and hematological improvement in these patients with acceptable side effects. 5-AZA is an attractive option for elderly patients with high-risk MDS.


Asunto(s)
Anemia Refractaria con Exceso de Blastos , Azacitidina , Médula Ósea , Esperanza de Vida , Síndromes Mielodisplásicos , Pancitopenia , Factores de Edad , Anciano , Anemia Refractaria con Exceso de Blastos/etiología , Anemia Refractaria con Exceso de Blastos/metabolismo , Anemia Refractaria con Exceso de Blastos/fisiopatología , Anemia Refractaria con Exceso de Blastos/terapia , Antimetabolitos Antineoplásicos/administración & dosificación , Antimetabolitos Antineoplásicos/efectos adversos , Azacitidina/administración & dosificación , Azacitidina/efectos adversos , Transfusión de Componentes Sanguíneos , Médula Ósea/patología , Médula Ósea/fisiopatología , Examen de la Médula Ósea , Humanos , Masculino , Monitoreo Fisiológico , Síndromes Mielodisplásicos/complicaciones , Síndromes Mielodisplásicos/metabolismo , Síndromes Mielodisplásicos/patología , Síndromes Mielodisplásicos/fisiopatología , Síndromes Mielodisplásicos/terapia , Pancitopenia/complicaciones , Pancitopenia/metabolismo , Pancitopenia/fisiopatología , Pancitopenia/terapia , Resultado del Tratamiento
9.
Exp Hematol ; 37(11): 1310-7, 2009 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-19748549

RESUMEN

OBJECTIVE: To investigate the contribution of cellular senescence to the progression and prognosis of myelodysplastic syndrome (MDS). MATERIALS AND METHODS: We have analyzed the expression of p16INK4a in bone marrow mononuclear cells or CD34(+) cells from 53 patients with MDS, 12 acute myeloid leukemia (AML), and 11 healthy controls. Additionally, We have assessed quantitatively senescence-associated beta-galactosidase (SA-beta-gal) staining on bone marrow mononuclear cells from MDS and AML patients, HL60 and SHI-1 leukemia cell lines, and healthy control cells. RESULTS: An upregulated expression of senescence-associated molecular marker p16INK4a was found in MDS compared with healthy controls, while a lower expression of p16INK4a was observed in AML compared with healthy controls. International Prognostic Scoring System score was negatively correlated with the percentage of p16INK4a-positive cells. The SA-beta-gal activity measured by mean percentage of positive cells was significantly higher in MDS cases when compared with controls. Meanwhile, percentage of SA-beta-gal-positive cells was also remarkably higher in dysplastic cells of MDS when compared to nondysplastic cells from MDS. CONCLUSIONS: These results of our present study suggested an accelerated cellular senescence occurred in MDS, and the cellular senescence may be involved in the progression and prognosis of MDS.


Asunto(s)
Senescencia Celular , Leucemia Mieloide/patología , Síndromes Mielodisplásicos/patología , Enfermedad Aguda , Adolescente , Adulto , Anciano , Anemia Refractaria/metabolismo , Anemia Refractaria/patología , Anemia Refractaria con Exceso de Blastos/metabolismo , Anemia Refractaria con Exceso de Blastos/patología , Biomarcadores , Línea Celular Tumoral/química , Línea Celular Tumoral/patología , Inhibidor p16 de la Quinasa Dependiente de Ciclina/análisis , Femenino , Genes p16 , Humanos , Leucemia Mieloide/metabolismo , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/metabolismo , Proteínas de Neoplasias/análisis , ARN Mensajero/análisis , ARN Neoplásico/análisis , Índice de Severidad de la Enfermedad , Adulto Joven , beta-Galactosidasa/análisis
10.
Blood ; 113(26): 6541-8, 2009 Jun 25.
Artículo en Inglés | MEDLINE | ID: mdl-19389880

RESUMEN

This study investigated the immunogenicity of Wilms tumor gene product 1 (WT1)-peptide vaccination in WT1-expressing acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) patients without curative treatment option. Vaccination consisted of granulocyte-macrophage colony-stimulating factor subcutaneously days 1 to 4, and WT1.126-134 peptide and 1 mg keyhole limpet hemocyanin on day 3. The initial 9 patients received 4 vaccinations biweekly, then monthly, and the subsequent 10 patients received continual biweekly vaccination. Seventeen AML patients and 2 refractory anemia with excess blasts patients received a median of 11 vaccinations. Treatment was well tolerated. Objective responses in AML patients were 10 stable diseases (SDs) including 4 SDs with more than 50% blast reduction and 2 with hematologic improvement. An additional 4 patients had clinical benefit after initial progression, including 1 complete remission and 3 SDs. WT1 mRNA levels decreased at least 3-fold from baseline in 35% of patients. In 8 of 18 patients, WT1-tetramer(+) T cells increased in blood and in 8 of 17 patients in bone marrow, with a median frequency in bone marrow of 0.18% at baseline and 0.41% in week 18. This WT1 vaccination study provides immunologic, molecular, and preliminary evidence of potential clinical efficacy in AML patients, warranting further investigations.


Asunto(s)
Anemia Refractaria con Exceso de Blastos/terapia , Vacunas contra el Cáncer/uso terapéutico , Inmunoterapia Activa , Leucemia Mieloide/terapia , Fragmentos de Péptidos/inmunología , Proteínas WT1/inmunología , Enfermedad Aguda , Adyuvantes Inmunológicos/administración & dosificación , Anciano , Anciano de 80 o más Años , Anemia Refractaria con Exceso de Blastos/genética , Anemia Refractaria con Exceso de Blastos/inmunología , Anemia Refractaria con Exceso de Blastos/metabolismo , Linfocitos T CD8-positivos/inmunología , Vacunas contra el Cáncer/inmunología , Eritema/etiología , Femenino , Genes del Tumor de Wilms , Humanos , Esquemas de Inmunización , Inmunoterapia Activa/efectos adversos , Inyecciones Intradérmicas , Inyecciones Subcutáneas , Leucemia Mieloide/genética , Leucemia Mieloide/inmunología , Leucemia Mieloide/metabolismo , Masculino , Persona de Mediana Edad , Proteínas de Neoplasias/inmunología , Fragmentos de Péptidos/administración & dosificación , Fragmentos de Péptidos/efectos adversos , ARN Mensajero/biosíntesis , ARN Neoplásico/biosíntesis , Terapia Recuperativa , Especificidad del Receptor de Antígeno de Linfocitos T , Proteínas WT1/administración & dosificación , Proteínas WT1/efectos adversos
11.
Genet Mol Res ; 7(4): 1245-50, 2008.
Artículo en Inglés | MEDLINE | ID: mdl-19065759

RESUMEN

The molecular pathogenesis of myelodysplastic syndromes (MDS) is poorly understood. In order to expand our knowledge of genetic defects in MDS, we determined the overall profile of genes expressed in bone marrow from patients with refractory anemia with excess blasts (RAEB) by serial analysis of gene expression (SAGE). The present report describes a partial transcriptome of RAEB bone marrow derived from 56,694 sequenced tags that provides information about expressed gene products. This is the first attempt to determine an overall profile of gene expression specifically in RAEB at diagnosis using SAGE, which should be useful in the understanding of the physiopathology of MDS and in identifying the genes involved.


Asunto(s)
Perfilación de la Expresión Génica/métodos , Expresión Génica , Síndromes Mielodisplásicos/genética , Adulto , Anciano , Anemia Refractaria con Exceso de Blastos/genética , Anemia Refractaria con Exceso de Blastos/metabolismo , Médula Ósea/metabolismo , Etiquetas de Secuencia Expresada , Femenino , Humanos , Masculino , Persona de Mediana Edad
13.
Pathol Int ; 57(10): 645-51, 2007 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-17803653

RESUMEN

Wilms' tumor 1 (WT1) is a useful marker for the diagnosis of acute leukemia and myelodysplastic syndromes (MDS). In the current study quantitative reverse transcription-polymerase chain reaction and immunostaining were used simultaneously to examine the relationship between WT1 RNA and protein level and also to evaluate WT1 as a tool to differentiate aplastic anemia (AA) and MDS refractory anemia (RA). Three types of WT1 messages (total, exon 5(+) and KTS(+)) and WT1 immunostaining of these diseases were analyzed. An increase of all three WT1 messages in high-grade MDS and acute leukemia was observed as compared with the normal control, whereas there was no significant difference in WT1 message between AA and RA, suggesting that WT1 message is not a good tool to discriminate AA and RA. No significant difference was observed between normal and RA, except for exon 5 message. Three WT1 message levels had a significant correlation, suggesting that the total WT1 message is sufficient for clinical practice. Positive immunostaining of WT1 was observed only in the portion of acute leukemia and overt leukemia (OL) transformed from MDS with a high WT1 message level, suggesting the relatively high detection threshold of WT1 protein with the immunostaining method.


Asunto(s)
Regulación de la Expresión Génica , Leucemia Mieloide Aguda/metabolismo , Síndromes Mielodisplásicos/metabolismo , ARN Mensajero/metabolismo , Proteínas WT1 , Adulto , Anciano , Anemia Aplásica/genética , Anemia Aplásica/metabolismo , Anemia Aplásica/patología , Anemia Refractaria con Exceso de Blastos/genética , Anemia Refractaria con Exceso de Blastos/metabolismo , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Diagnóstico Diferencial , Femenino , Humanos , Técnicas para Inmunoenzimas , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/genética , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/diagnóstico , Síndromes Mielodisplásicos/genética , ARN Neoplásico/análisis , Proteínas WT1/genética , Proteínas WT1/metabolismo
14.
Br J Haematol ; 138(5): 644-50, 2007 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-17686059

RESUMEN

A-kinase anchor protein 12 (AKAP12) is a scaffold protein that participates in mitotic regulation and other signalling processes and probably exerts tumour suppressor function. We hypothesized that epigenetic repression of the AKAP12 gene might occur in malignant myeloid disorders. This study demonstrated that the 5' CpG island of AKAP12 was unmethylated in normal haematopoietic progenitors and granulocytes but exhibited profound methylation in Kasumi-1 and SKNO-1 leukaemic myeloblasts. Correspondingly, AKAP12 was expressed in normal progenitors but transcriptionally silent in leukaemic blasts. Re-expression of AKAP12 in Kasumi-1 and SKNO-1 cells was accomplished by treatment with MS275 alone or in combination with zebularine, indicating epigenetic mechanisms of gene repression. AKAP12 hypermethylation was found in one case of refractory anaemia with excess blasts (RAEB) and two cases of acute myeloid leukaemia (AML) in a panel of 21 blood or bone marrow samples from children with malignant myeloid disorders including refractory cytopenia, RAEB, juvenile myelomonocytic leukaemia and AML. While AKAP12 function has not been previously linked to leukaemogenesis, our results raise the possibility that epigenetic silencing of AKAP12 is involved in myeloid malignancies.


Asunto(s)
Proteínas de Ciclo Celular/genética , Metilación de ADN , ADN de Neoplasias/genética , Leucemia Mieloide/genética , Síndromes Mielodisplásicos/genética , Proteínas de Anclaje a la Quinasa A , Adolescente , Anemia Refractaria con Exceso de Blastos/genética , Anemia Refractaria con Exceso de Blastos/metabolismo , Proteínas de Ciclo Celular/metabolismo , Niño , Preescolar , Epigénesis Genética , Femenino , Silenciador del Gen , Humanos , Lactante , Recién Nacido , Leucemia Mieloide/metabolismo , Leucemia Mielomonocítica Aguda/genética , Leucemia Mielomonocítica Aguda/metabolismo , Masculino , Síndromes Mielodisplásicos/metabolismo , Proteínas de Neoplasias/metabolismo , Regiones Promotoras Genéticas , ARN Mensajero/genética , ARN Neoplásico/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Células Tumorales Cultivadas
16.
Am J Clin Pathol ; 125(6): 884-94, 2006 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-16690488

RESUMEN

We used a new method of data presentation and analysis, termed antigen mapping, to characterize recurring myeloblast phenotypic abnormalities in a series of 28 cases of myelodysplastic syndrome (MDS), including refractory anemia with ringed sideroblasts (RARS), refractory anemia with multilineage dysplasia (RCMLD), and refractory anemia with excess blasts (RAEB). Abnormal patterns of CD34 and CD117 expression were present in 50% of RARS, 68% of RCMLD, and 100% of RAEB cases. The presence of decreased myeloblast CD45 density, increased CD13 and CD34 density, and increased expression of CD11c and CD4(dim) were MDS grade-related. There was a direct relationship between the number of myeloblast phenotypic abnormalities (phenotypic score) and MDS grade. The myeloblast phenotypic scores also were correlated highly with International Prognostic Scoring System scores and risk categories. We found the antigen mapping technique to be an efficient data presentation and analysis method for the detection of MDS-associated abnormalities of antigen distribution and density.


Asunto(s)
Anemia Refractaria con Exceso de Blastos/metabolismo , Antígenos CD34/metabolismo , Médula Ósea/metabolismo , Células Precursoras de Granulocitos/metabolismo , Proteínas Proto-Oncogénicas c-kit/metabolismo , Anciano , Anciano de 80 o más Años , Anemia Refractaria con Exceso de Blastos/patología , Biomarcadores/metabolismo , Médula Ósea/patología , Mapeo Epitopo , Femenino , Células Precursoras de Granulocitos/patología , Humanos , Inmunofenotipificación , Masculino , Persona de Mediana Edad , Fenotipo
17.
Leuk Lymphoma ; 47(3): 451-60, 2006 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-16396768

RESUMEN

Recent data suggest that vascular endothelial growth factor (VEGF) is produced in neoplastic cells in various myeloid neoplasms and plays a key role as an autocrine regulator and mediator of angiogenesis. We examined the expression of VEGF in paraffin-embedded bone marrow sections obtained from normal donors (n = 5) and 46 patients with myelodysplastic syndromes [MDS, French-American-British (FAB)-type refractory anemia (RA), n = 10; refractory anemia with ringed sideroblasts (RARS), n = 10; refractory anemia with excess blasts (RAEB), n = 10; RAEB in transformation (RAEB-T), n = 8; chronic myelomonocytic leukemia (CMML), n = 8] by immunohistochemistry using an anti-VEGF antibody. In normal bone marrow, the anti-VEGF antibody was found to react with myeloid progenitor cells, immature monocytic cells, plasma cells and megakaryocytes, but not with erythroid cells or mature granulocytic cells. Higher levels of VEGF were found in patients with MDS, subtypes RAEB, RAEB-T and CMML, compared to patients with RA or RARS, or the normal bone marrow. These differences were found to result from expression of VEGF in immature myeloid cells in RAEB, RAEB-T and CMML. The microvessel density was also higher in patients with RAEB-T and CMML compared to RA and RARS or the normal bone marrow. Expression of VEGF mRNA was demonstrable in isolated neoplastic cells by reverse transcriptase-polymerase chain reaction in all patients examined. In aggregate, these data show that VEGF is expressed in bone marrow cells in patients with MDS. The amount of expressed VEGF is related to the percentage of immature myeloid cells (blasts and monocytic progenitors) and correlates with the FAB category.


Asunto(s)
Médula Ósea/patología , Síndromes Mielodisplásicos/clasificación , Síndromes Mielodisplásicos/patología , Factores de Crecimiento Endotelial Vascular/biosíntesis , Factores de Crecimiento Endotelial Vascular/genética , Adulto , Anciano , Anciano de 80 o más Años , Anemia Refractaria con Exceso de Blastos/metabolismo , Anemia Refractaria con Exceso de Blastos/patología , Médula Ósea/irrigación sanguínea , Médula Ósea/metabolismo , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/metabolismo , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Valor Predictivo de las Pruebas , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Factores de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Receptores de Factores de Crecimiento Endotelial Vascular/genética , Receptores de Factores de Crecimiento Endotelial Vascular/metabolismo
18.
Anal Quant Cytol Histol ; 27(3): 152-6, 2005 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-16121636

RESUMEN

OBJECTIVE: To study the histomorphometric features of megakaryocytic elements in bone marrow trephine biopsies of various subtypes of myelodysplastic syndrome (MDS). STUDY DESIGN: Comparative image morphometry using hematoxylin-eosin-stained and CD 61-stained trephine biopsies was carried out on 40 cases of MDS and 10 normal subjects to analyze the megakaryocytes objectively. The various variables analyzed were number of megakaryocytes and micromegakaryocytes, area, perimeter and diameter of the megakaryocytic elements. RESULTS: The mean number of megakaryocytes was lower in cases of MDS as compared to the normals in all except for a single case of hypoplastic MDS, in which the megakaryocytes were more abundant (3.6 per high-power field [hpf]). No micromegakaryocytes were observed in the 2 cases of chronic myelomonocytic leukemia. The mean area, perimeter and diameter of megakaryocytes decreased significantly on immunostaining with CD 61. CONCLUSION: The mean number of megakaryocytes per hpf was lower in the cases of MDS as compared to normal cases on hematoxylin-eosin. However, on CD 61 staining the number of megakaryocytes per hpf increased in cases of MDS. Micromegakaryocytes were seen in scanty numbers in the normals but increased in MDS cases and increased significantly on CD 61 immunostaining. The mean area, perimeter and diameter of megakaryocytes decreased significantly on immunostaining with CD 61, indicating the increased numbers of micromegakaryocytes in MDS. Hence, immunostaining is an efficient method of detecting increased numbers of megakaryocytes and micromegakaryocytes that would ordinarily be missed by using routine hematoxylin-eosin staining.


Asunto(s)
Células de la Médula Ósea/patología , Citometría de Imagen/métodos , Síndromes Mielodisplásicos/patología , Anemia Aplásica/metabolismo , Anemia Aplásica/patología , Anemia Refractaria/metabolismo , Anemia Refractaria/patología , Anemia Refractaria con Exceso de Blastos/metabolismo , Anemia Refractaria con Exceso de Blastos/patología , Anemia Sideroblástica/metabolismo , Anemia Sideroblástica/patología , Biopsia , Células de la Médula Ósea/química , Recuento de Células , Histocitoquímica , Humanos , Inmunohistoquímica , Integrina beta3/análisis , Integrina beta3/inmunología , Leucemia Mielomonocítica Crónica/metabolismo , Leucemia Mielomonocítica Crónica/patología , Megacariocitos/química , Megacariocitos/patología , Síndromes Mielodisplásicos/metabolismo , Estadísticas no Paramétricas
19.
Chin Med J (Engl) ; 118(7): 548-54, 2005 Apr 05.
Artículo en Inglés | MEDLINE | ID: mdl-15820085

RESUMEN

BACKGROUND: The inhibitor of apoptosis (IAP) gene family is involved in the suppression of apoptotic cell death as well as an increasing number of seemingly unrelated cellular functions. It is not known, however, whether IAP expression in malignant hematopoietic cells is affected by chemotherapeutic agents such as homoharringtonine (HHT). In this study, we investigated mRNA expression levels of IAPs, especially survivin, in various hematopoietic cell lines in relation with apoptosis induced by HHT. METHODS: Semiquantitative reverse transcriptase polymerase chain reaction was used to determine survivin mRNA levels. Cell apoptosis was examined by flow cytometry. Cell viability and proliferation assay was evaluated by MTT. The experiments were performed on the malignant hematopoietic cell lines MUTZ-1, K562, Jurkat, RMPI and HL60, with or without survivin antisense-oligodeoxynucleotides (AS-ODN) and HHT. RESULTS: The expression levels of survivin mRNA were variable in the cell lines and negatively correlated to HHT induced cell apoptosis. Survivin AS-ODN significantly decreased mRNA level of survivin, but not those of bax and bcl-2. Survivin also inhibited MUTZ-1 cell growth and induced apoptosis in a dose dependent manner. AS-ODN and HHT showed synergistic effect on MUTZ-1 cell growth. CONCLUSION: The apoptotic effect of HHT on the hematopoietic cell lines is associated with decreased level of survivin expression. Survivin could be a new marker for drug sensitivity and a new target for cancer treatment.


Asunto(s)
Anemia Refractaria con Exceso de Blastos/patología , Apoptosis/efectos de los fármacos , Harringtoninas/farmacología , Leucemia/patología , Proteínas Asociadas a Microtúbulos/genética , Anemia Refractaria con Exceso de Blastos/metabolismo , Ciclo Celular , Línea Celular , Homoharringtonina , Humanos , Proteínas Inhibidoras de la Apoptosis , Leucemia/metabolismo , Proteínas de Neoplasias , Oligonucleótidos Antisentido/farmacología , Proteínas Proto-Oncogénicas c-bcl-2/genética , ARN Mensajero/análisis , Survivin , Proteína X Asociada a bcl-2
20.
Leuk Res ; 27(7): 583-91, 2003 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-12681357

RESUMEN

Myelodysplastic syndromes (MDS) are characterized by peripheral blood cytopenias despite hypercellularity of the bone marrow regarded as the result of ineffective hematopoiesis mainly caused by apoptosis. In this study, we examined the role of tumor necrosis factor (TNF)-induced apoptosis in the bone marrow cells of MDS patients. The constitutive expression of mRNA for TNF receptors (TNFR), including TNFRI and TNFRII, and the adapter molecules, such as the TNF receptor-associated death domain protein (TRADD), Fas-associated death domain protein (FADD), receptor interacting protein (RIP) and TNF receptor-associated factor 2 (TRAF-2) were analyzed by reverse transcriptase (RT)-PCR in bone marrow samples from control, MDS and AML cases. In bone marrow cells from refractory anemia (RA) patients, there was a significant increase in TNFRI expression as compared with control subjects. The expression of TNFRII was also up-regulated in RA cases. In contrast, RA with excess of blasts (RAEB), RAEB in transformation (RAEB-T) and AML cases revealed increased expression of TNFRII, whereas the expression of TNFRI was comparable to control subjects. Immunohistochemical staining revealed that the TNFRI, as well as TNFRII of MDS bone marrow was expressed mainly in hematopoietic cells, but not in macrophage-lineage stromal cells at the protein level. An increased constitutive expression of mRNA for TRADD, FADD and RIP and decreased expression of mRNA for TRAF-2 were observed in bone marrow cells from MDS patients, especially from RA patients, as compared with controls, although the differences were not significant. In many of the AML bone marrow samples, strong expression of TRAF-2 mRNA was marked, while expression levels of other proteins were similar to those in control subjects. These data suggested enhanced signaling by the TNFRI-TRADD-FADD pathway and suppressed signaling by the TRAF-2 pathway in RA. Thus, TNF-alpha-induced apoptosis may play a role in ineffective hematopoiesis in "early stage MDS" bone marrow, although the regulatory mechanisms for TNF-alpha-induced signaling would be complicated and not be simply explained only by these pathways.


Asunto(s)
Antígenos CD/genética , Células de la Médula Ósea/metabolismo , Regulación Neoplásica de la Expresión Génica , Síndromes Mielodisplásicos/genética , Receptores del Factor de Necrosis Tumoral/genética , Transducción de Señal , Adulto , Anciano , Anciano de 80 o más Años , Anemia Refractaria/metabolismo , Anemia Refractaria/patología , Anemia Refractaria con Exceso de Blastos/metabolismo , Anemia Refractaria con Exceso de Blastos/patología , Antígenos CD/metabolismo , Apoptosis/efectos de los fármacos , Células de la Médula Ósea/patología , Regulación hacia Abajo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas/genética , Proteínas/metabolismo , ARN Mensajero/metabolismo , Proteína Serina-Treonina Quinasas de Interacción con Receptores , Receptores del Factor de Necrosis Tumoral/metabolismo , Receptores Tipo I de Factores de Necrosis Tumoral , Receptores Tipo II del Factor de Necrosis Tumoral , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor 1 Asociado a Receptor de TNF , Factor 2 Asociado a Receptor de TNF , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Regulación hacia Arriba , Receptor fas/genética , Receptor fas/metabolismo
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