Effect of MiR-142-3p Targeting HOXA5 on Proliferation, Cycle Arrest and Apoptosis of Acute B Lymphocytic Leukemia Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect and molecular mechanism of miR-142-3p to the proliferation, cycle and apoptosis of acute B lymphocytic leukemia (B-ALL) cells by regulating the homeobox gene 5 (HOXA5) expression.@*METHODS@#Real-time fluorescence quantitative PCR was used to detect the expression levels of miR-142-3p and HOXA5 in human B-ALL cell Nalm6 cell line and human B lymphoblast Hmy2-cir cells. Nalm6 was transfected by using liposome transfection technology, miR-142-3p mimic, pcDNA-HOXA5 overexpression plasmid, miR-142-3p mimic+pcDNA-HOXA5 overexpression plasmid, and control. The binding site of HOXA5 and miR-142-3p was predicted according to microRNA.org, and the targeting relationship between miR-142-3p and HOXA5 gene was detected by double luciferase reporter gene experiment. The effect of miR-142-3p to the proliferation of Nalm6 cells was detected using the Cell Counting Box-8 (CCK-8) method and cell clone formation experiments. Flow cytometry was used to detect the effects of miR-142-3p to cell cycle distribution and apoptosis of Nalm6 cells. The expression levels of cell cycle-related proteins, including G@*RESULTS@#Compared with Hmy2-cir cells, miR-142-3p showed low expression in Nalm6 cells and HOXA5 showed high expression (P<0.05). MiR-142-3p and HOXA5 3'-UTR showed complementary binding regions, the luciferase activity of miR-142-3p mimic and wild-type HOXA5 3'-UTR was significantly lower than that of miR-142-3p negative control and wild-type HOXA5 3'-UTR (P<0.05). The proliferation of Nalm6 cells and the number of cell clones could be inhibited by miR-142-3p mimic after 48 and 72 hours of transfection (P<0.05), which causing G@*CONCLUSION@#MiR-142-3p can inhibit the proliferation of Nalm6 cells by targeting down-regulation the expression of HOXA5, arrest the G

Sobrevida global de pacientes con leucemia aguda en el Hospital de Especialidades Pediátricas de Chiapas, México / Childhood acute leukemia overall survival at the Hospital de Especialidades Pediátricas in Chiapas, México

Resumen: Introducción: A 10 años de la fundación del Hospital de Especialidades Pediátricas en Chiapas, México, es importante valorar la sobrevida global a 5 años de los pacientes con leucemia aguda bajo el régimen del Seguro Popular. Métodos: Estudio descriptivo y de sobrevida de 210 casos de leucemia aguda diagnosticados y tratados entre 2008 y 2012. Empleando curvas de Kaplan-Meier se analizó cada variedad de la enfermedad (B, T y mieloide), y para la leucemia B en función del grupo de riesgo, el sexo, la edad, los leucocitos al diagnóstico, los marcadores de superficie, el índice de DNA, el cariotipo y las translocaciones. Resultados: La edad, el sexo y la proporción de tipos de leucemia aguda (B = 85%; M = 10%; T = 5%) fueron similares al resto del país. El 20% de los pacientes estaban vivos a 5 años; el 53% habían fallecido y el 27% abandonaron el tratamiento. La sobrevida global a 5 años fue del 42% (B = 45%; T = 20%; M = 10%) (mediana: 38.8 meses; intervalo de confianza del 95%: 28.9-48.7). La mediana de «muy alto riesgo¼ fue de 7.7 contra 47 meses; no hubo diferencia entre riesgo habitual y alto riesgo. Los leucocitos < 50,000/µl al diagnóstico y CD10 positivo se asociaron con mejor sobrevida. En el momento del deceso, el 29% se encontraba en remisión. Conclusiones: La sobrevida de la leucemia aguda bajo el Seguro Popular fue desfavorable los primeros 5 años del Hospital de Especialidades Pediátricas. Se identificaron como contribuyentes la alta tasa de mortalidad temprana, de pacientes en remisión y el abandono. Además de revisar la atención médica, se requiere el estudio de elementos extrahospitalarios determinantes del abandono para mejorar el programa.

Abstract: Background: At the 10th anniversary of the Hospital de Especialidades Pediátricas in Chiapas, Mexico, it was important to assess the 5-year acute leukemia overall survival under the Seguro Popular program (Popular Insurance). Methods: A descriptive and survival study of 210 acute leukemia patients diagnosed and treated during 2008-2012 was performed. Kaplan-Meier survival curves were developed for all patients, each leukemia type (B, T and myeloid) and for B type related to risk group, age, sex, leukocytes, cell markers, DNA index, karyotype, and translocations. Results: Age, gender and proportion of leukemia types (B = 85%; M = 10%; T = 5%), were similar to other parts of the country. At the end of the 5-year treatment, 20% of the patients were alive, 53% had died and 27% had abandoned the treatment. Global survival was 42% (B = 45%; T = 20%; M = 10%) (median: 38.8 months; confidence interval of 95% = 28.9-48.7). Very high-risk median survival was 7.7 versus 47 months. There was no difference between standard and high-risk groups. The initial leukocyte count < 50,000/µL and CD10 positive were related to better B survival; no other variables were related. At the time of death, 29% of patients were in remission. Conclusions: Global survival of acute leukemia at Hospital de Especialidades Pediátricas under the Seguro Popular during its first 5 years was surprisingly poor given the medical resources available through the insurance. Early mortality, death during remission and high desertion rates contributed to these results. A detailed revision of treatment protocols and reasons for abandoning treatment is mandatory.

Correlación de la t(9;22), t(12;21) e hiperdiploidía de ADN con el inmunofenotipo y la tasa de proliferación de células B neoplásicas en niños con leucemia linfoblástica aguda de precursores / Correlation of the t(9;22), t(12;21), and DNA hyperdiploid content with immunophenotype and proliferative rate of leukemic B-cells of pediatric patients with B-cell acute lymphoblastic leukemia

Introducción. Del 60 al 80 % de los pacientes con leucemia linfoblástica aguda de precursores B presentan alteraciones genéticas que influyen en el pronóstico de la enfermedad y en la biología del tumor. Objetivo. Analizar distintas alteraciones genéticas en leucemia linfoblástica aguda de precursores B en niños, y su relación con el inmunofenotipo y con la tasa de proliferación, en comparación con precursores B normales. Materiales y métodos. En 44 pacientes se evaluó, por citometría de flujo, el inmunofenotipo, el contenido de ADN y la proliferación, y por RT-PCR, las traslocaciones t(9;22), t(12;21), t(4;11) y t(1;19). Mediante un análisis jerarquizado de conglomerados se identificaron los patrones inmunofenotípicos de expresión asociados a las traslocaciones, tomando como referencia precursores B normales. Resultados. La cuantificación del ADN mostró que el 21 % de los casos de leucemia linfoblástica aguda de precursores B eran hiperdiploides de índice alto y, el 47,7 %, hiperdiploides de índice bajo. La presencia de hiperdiploidía se asoció con mayor proliferación tumoral y con inmunofenotipos aberrantes, que incluyeron expresión anormal de CD10, TdT, CD38 y CD45 y un mayor tamaño de los linfoblastos. La presencia de t(9;22) y t(12;21) discrimina células normales de células tumorales con aberraciones en la expresión de CD19, CD20, CD13, CD33, CD38, CD34 y CD45. Conclusiones. El perfil de aberraciones fenotípicas detectado en conjunto con anormalidades en la proliferación tumoral, se asocia de forma significativa con hiperdiploidiía de ADN y discrimina de forma clara linfoblastos con t(9;22) y t(12;21) de los precursores B normales. La identificación de estos parámetros será de gran utilidad como herramienta para la clasificación y seguimiento de los pacientes.

Introduction: Between 60 and 80% of patients with B-cell acute lymphoblastic leukemia show genetic abnormalities which influence the prognosis of the disease and the biology of the tumor. Objective: To analyze different genetic abnormalities in acute B lymphoblastic leukemia in children, its relationship with the immunophenotype and the proliferative rate compared with normal B cell precursors. Materials and methods: We assessed immunophenotype, DNA content and proliferative rate in 44 samples by flow cytometry, and translocations t(9;22), t(12;21), t(4;11), and t(1;19) by RT-PCR. Using a hierarchical cluster analysis, we identified some immunophenotypic patterns associated to genetic abnormalities when compared with normal B cell precursors. Results: DNA quantification showed that 21% of the cases had high hyperdiploidy and 47.7% has low hyperdiploidy. The presence of hyperdiploidy was associated with increased tumor proliferation and aberrant immunophenotypes, including abnormal expression of CD10, TdT, CD38, and CD45 and an increased size of the lymphoblasts. The presence of t(9;22) and t(12;21) discriminates normal cells from tumor cells with aberrant immunophenotype in the expression of CD19, CD22, CD13, CD33, CD38, CD34, and CD45. Conclusions: The aberrant immunophenotype profile detected in neoplastic cells along with abnormalities in the proliferative rate were significantly associated with DNA hyperdiploidy and clearly distinguished lymphoblasts with t(9;22) and t(12;21) from normal B cell precursors. The identification of these parameters is useful as a tool for classification and monitoring of these patients.

Immunophenotypic characterization of the leukemic B-cells from Iranian patients with chronic lymphocytic leukemia: association between CD38 expression and disease progression

Patients with B-cell chronic lymphocytic leukemia [B-CLL] have heterogeneous clinical courses, thus several biological parameters need to be added to the current clinical staging systems to predict disease outcome. Recent immunophenotypic studies performed mainly in Western populations have demonstrated the prognostic value of CD38 and ZAP-70 expression in B-CLL. To investigate the expression pattern of a variety of membrane antigens on leukemic cells from Iranian patients with CLL and to find out if there are any differences in the expression of these markers between indolent and progressive groups. In the present study, peripheral blood samples from 87 Iranian patients with B-CLL were analysed by flow cytometry. In all cases, the neoplastic cells displayed B-CLL phenotype [CD5[+]/CD19[+]/sIg[+]]. The vast majority of the cases expressed CD23, but failed to stain for CD3 or CD14. The leukemic cells of most patients expressed CD27 [84/87, 95.4%] and CD45RO [74/87, 83.9%] molecules, suggesting a memory B-cell phenotype. Comparison between the indolent [n=42] and progressive [n=37] patients revealed significantly higher frequency and intensity of CD38 expression in progressive group [40.5%] compared to indolent [11.9%] patients [p<0.05]. None of the other membrane antigens were differentially expressed in these two groups of patients. Our results obtained in an Asian ethnic population confirm and extend previous findings obtained from Western populations regarding the association of CD38 expression and disease progression in B-CLL

Leucemia de células peludas / Hairy cell leukemia

Se describen 4 casos de pacientes con leucemia de células peludas en dos generaciones de una misma familia; se comenta el tratamiento y la evolución de los casos, enfatizando el papel de la herencia en las leucemias