Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 88
Filter
1.
Article in Chinese | WPRIM | ID: wpr-880129

ABSTRACT

OBJECTIVE@#To investigate the expression of WTAP gene in acute myeloid leukemia (AML) and its clinical significance.@*METHODS@#74 acute myeloid leukemia patients with non-M3 type and 19 normal donors were selected, and real-time quantitative polymerase chain reaction was used to detect the mRNA expression level of WTAP gene in their bone marrow cells. The relationship between the mRNA expression level of WTAP gene and the clinical characteristics was analyzed.@*RESULTS@#The relative mRNA expression of WTAP gene in the non-M3 AML group was significantly higher than that in the healthy control group, and the difference showed statistically significant (P0.05) according to the classification of FAB. The mRNA expression level of WTAP gene in FLT3-ITD mutated AML patients was higher than that in FLT3-ITD unmutated group (P=0.016), and the mRNA expression level of WTAP gene in AML patients with CEBPα mutation was lower than that in CEBPα unmutated group (P=0.016). The expression level of WTAP mRNA was positively correlated with WT1 expression (r=0.6866, P0.05). The expression level of WTAP mRNA showed no obvious effect on the complete remission of patients after first treatment. The different expression level of WTAP gene at initial diagnosis showed also no effect on the overall survival time of patients.@*CONCLUSION@#The expression level of WTAP gene is increasing in new diagnosed non-M3 acute myeloid leukemia. There is a positive correlation between the expression level of WTAP gene and the expression level of WT1 fusion gene. WTAP mRNA always shows higher expression in patients with FLT3-ITD mutation than that in patients without FLT3-ITD mutation, and shows lower expression in patients with CEBPα mutation than that in unmutated group.


Subject(s)
Cell Cycle Proteins , Humans , Karyotype , Leukemia, Myeloid, Acute/genetics , Mutation , Prognosis , RNA Splicing Factors , Remission Induction , fms-Like Tyrosine Kinase 3/genetics
2.
Article in Chinese | WPRIM | ID: wpr-880096

ABSTRACT

OBJECTIVE@#To investigate the relationship between acute myeloid leukemia (AML) patients ASXL2, ZBTB7A gene mutations and the prognosis.@*METHODS@#42 AML Patients treated in our hospital from January 2014 to January 2016 were selected and ASXL2 and ZBTB7A genes of their bone marrow samples were sequenced, the genetic characteristics and prognosis of core-binding factor-AML(CBF-AML) patients with ASXL2 and ZBTB7A mutations were analyzed.@*RESULTS@#ASXL2 (33.3%) and ZBTB7A (9.5%) mutations were found in t (8; 21) AML patients. Compared with wild-type, patients with ASXL2 mutations showed significantly higher white blood cell count at diagnosis [(9.49±1.85)×10@*CONCLUSION@#ASXL2 and ZBTB7A mutations are frequently found in t (8; 21) AML patients. The mutation of ASXL2 and ZBTB7A genes shows no significant effect on the prognosis of AML patients.


Subject(s)
Cell Line, Tumor , DNA-Binding Proteins/genetics , Humans , Leukemia, Myeloid, Acute/genetics , Mutation , Oncogene Proteins, Fusion/genetics , Prognosis , Repressor Proteins/genetics , Transcription Factors/genetics
3.
Article in Chinese | WPRIM | ID: wpr-880095

ABSTRACT

OBJECTIVE@#To investigate the expression of microRNA-370 (miR-370) and microRNA-203 (miR-203) in the serum of patients with acute myeloid leukemia(AML), and to analyze its clinical diagnosis and prognostic significance.@*METHODS@#57 patients with acute myeloid leukemia were enrolled as experimental group, and 21 healthy people were enrolled as control group. The fasting venous blood of the personal in the two groups were collected. The expression of miR-370 and miR-203 of the personal in each groups were detected by real-time fluorescent quantitative PCR. The receiver operating characteristic (ROC) curve was plotted to detected the diagnostic values of serum miR-370, miR-203, and the Kaplan-Meier method was used to estimate the relationship between expression and overall survival of the patients.@*RESULTS@#Compared with healthy controls, serum miR-370 expression was significantly decreased in AML patients(P<0.05), and serum miR-203 expression was also significantly decreased (P<0.05). ROC curve analysis showed that the expression of serum miR-370 and miR-203 could be used to distinguish acute myeloid leukemia and healthy people. The area under the ROC curve of miR-370 was 0.909, and the sensitivity and specificity were 91.46% and 100.00%, respectively. The area under the ROC curve of miR-203 was 0.895, and the sensitivity and specificity were 83.45% and 89.71%, respectively. Serum levels of miR-370 and miR-203 were closely related to overall survival in AML patients.@*CONCLUSION@#The expression of miR-370 and miR-203 is decreased in the serum of patients with AML and may be a new markers for the diagnosis and prognosis of AML.


Subject(s)
Biomarkers, Tumor , Humans , Kaplan-Meier Estimate , Leukemia, Myeloid, Acute/genetics , MicroRNAs , Prognosis , ROC Curve
4.
Article in Chinese | WPRIM | ID: wpr-880094

ABSTRACT

OBJECTIVE@#To analyze the relationship of the expression of transcription factor MYB targeted regulation by miR-96 to cell invasion and apoptosis in pediatric acute myeloid leukemia (AML).@*METHODS@#A total of 65 children with AML in The 928 Hospital of PLA Joint Logistics Support Forces from January 2017 to November 2019 were selected, including 35 cases diagnosed as primary AML and 30 cases as complete remission AML. Thirty children with immune thrombocytopenia were selected as control group. The clinical characteristics were analyzed and compared between the two groups. The levels of miR-96 and MYB in peripheral blood samples were detected by qRT-PCR and compared between the two groups. The miR-96 mimics and its negative control (NC), inhibitor-miR-96 and its NC transfected HL60 cells induced by liposome (Lipofectamine 2000), respectively, Then the expression levels of MYB were detected with Western blot and compared among four HL60 cell groups. The invasion ability of four HL60 cell groups were detected with Transwell assay. The cell proliferation ability of four HL60 cell groups were detected with MTT at 24 h, 48 h, and 72 h, respectively. The apoptosis rates of four HL60 cell groups were detected with flow cytometry.@*RESULTS@#Compared with control group, the level of miR-96 in AML children were higher, but MYB lower (P0.05). The promotion of over-expression level of miR-96 on the invasion ability of HL 60 cells was confirmed by Transwell assay. MTT assay showed that miR-96 could promote the proliferation of HL60 cells, inhibit the apoptosis of HL60 cells, and the effect was time-dependent manner (r=0.804). The inhibition of miR-96 on HL60 cells apoptosis was also confirmed with flow cytometry.@*CONCLUSION@#MiR-96 has significant negative effect on invasion and apoptosis of AML cells by targeting regulation MYB, and it might be a potential novel strategy for pediatric AML treatment.


Subject(s)
Apoptosis , Cell Line, Tumor , Cell Proliferation , Child , HL-60 Cells , Humans , Leukemia, Myeloid, Acute/genetics , MicroRNAs/genetics , Proto-Oncogene Proteins c-myb
5.
Article in Chinese | WPRIM | ID: wpr-880090

ABSTRACT

OBJECTIVE@#To investigate the expression of CD73 in acute myeloid leukemia (AML) patients with NPM1 mutant and wild-type, and to evaluate the therapeutic efficacy and prognosis of CD73 to the AML patients.@*METHODS@#160 patients with AML treated in our hospital from June 2015 to June 2019 were enrolled, and 40 non-AML bone marrow samples from healthy people were selected as controls during the same period. The expression of CD73 in healthy people, NPM1 mutation and NPM1 wild-type AML patients were compared, and the relationship between the expression of CD73 and its clinicopathological characteristics, as while as efficacy in AML patients were analyzed. The patients were followed up, and the influence of CD73 to the prognosis of different AML patients was analyzed.@*RESULTS@#The positive expression rate of CD73 in AML patients (23.75%) was significantly higher than that in the healthy control group (0.62%), and the positive expression rate of CD73 in AML patients with NPM1 mutation (74.75%) was significantly higher than that with NPM1 wild-type (25.51%) (both P<0.001). AML patients with CD73 positive expression was associated with age, FAB typing, disease risk classification, and NPM1 gene mutation (both P<0.05). The overall survival rate of AML patients with NPM1 gene mutation was 75.98%, which was significantly higher than the patients with NPM1 wild-type (34.68%)(P<0.001), the median survival time of AML patients with NPM1 gene mutation in the CD73@*CONCLUSION@#The expression of CD73 was increased in AML patients with NPM1 gene mutation, and CD73 showed different prognostic significance in AML patients with different NPM1 gene mutation. The combination of clinicopathologic features, CD73 expression and NPM1 gene in AML patients is helpful to determine their prognosis and guide the formulation of relevant treatment plans.


Subject(s)
Humans , Leukemia, Myeloid, Acute/genetics , Mutation , Nuclear Proteins/genetics , Prognosis , Survival Rate , fms-Like Tyrosine Kinase 3
6.
Article in Chinese | WPRIM | ID: wpr-880088

ABSTRACT

OBJECTIVE@#To establish a method for rapid detection and typing of NPM1 mutations in acute myeloid leukemia (AML) by fluorescence melting curve analysis technology.@*METHODS@#A pair of primers and a fluorescent single-stranded probe (molecule beacon) were designed for the mutant genes mutA, mutB, mutD in exon 12 of nucleopsin (NPM1) and wild type. With a real-time qPCR, the A, B, and D gene mutations of NPM1 were detected and typed by different-melting curve peak value of the probe through RT-PCR.@*RESULTS@#This method could detected the mutations of A, B, and D in NPM1 effectively with a sensitivity of 1%. Furthermore, 62 AML clinical samples were evaluated by the method. In the results, the detection rate and typing of NPM1 mutations were consistent with the sequencing results of clinical samples.@*CONCLUSION@#There are three features in the method of fluorescence melting curve analysis: stable PCR system, easy to operate, and the easily distinguishable results. The method might meet the demand for rapid typing of NPM1 gene mutation in early diagnosis or concomitant diagnosis of AML.


Subject(s)
Exons , Genotype , Humans , Leukemia, Myeloid, Acute/genetics , Mutation , Nuclear Proteins/genetics
7.
Article in Chinese | WPRIM | ID: wpr-880083

ABSTRACT

OBJECTIVE@#To analyze the prognostic factors of AML children with CBFβ/MYH11 positive.@*METHODS@#Twenty-eight children with CBFβ/MYH11 positive treated in our hospital from May 2012 to June 2018 were selected, the clinical data and curative were analyzed and evaluated.@*RESULTS@#Five-year OS and 5-year EFS rate of CBFβ/MYH11 positive AML children was 76.8% and 64.0% efficacy, respectively. Univariate analysis results showed that the OS rate of CBFβ/MYH11 positive AML children with WBC<60.0×10@*CONCLUSION@#WBC level and XRCC-Thr241Met genotype at initial diagnosis are the major affecting factors for prognosis of AML children with CBFβ/MYH11 positive.


Subject(s)
Child , Chromosome Inversion , Genotype , Humans , Leukemia, Myeloid, Acute/genetics , Myosin Heavy Chains , Oncogene Proteins, Fusion , Prognosis
8.
Article in Chinese | WPRIM | ID: wpr-880081

ABSTRACT

OBJECTIVE@#To investigate the mutational spectrum and its prognostic significance in patients with acute myeloid leukemia (AML).@*METHODS@#The clinical data of 93 patients with newly diagnosed AML who underwent gene mutation detection by high-throughput sequencing (HTS) from March 2014 to April 2018 in our hospital was analyzed retrospectively. The distribution of mutant genes was summarized and the prognostic factors for intermediate-risk acute myeloid leukemia (IR-AML) were analyzed.@*RESULTS@#Among 93 AML patients, 88.17% had at least one gene mutation, and 53.76% patients showed more than one recurrent genetic mutation. CEBPA showed the highest mutation frequency (20.4%), followed by ASXL1, TET2, NRAS, FLT3-ITD, NPM1, IDH2, DNMT3A, and their mutation frequency were higher than 10%. IDH1/2 and NPM1, ASXL1 and U2AF1, FLT3 and NPM1 often co-occured (P 100×10@*CONCLUSION@#There are co-occurring mutation patterns between the mutated genes. IDH2 mutations relates with poor prognosis and possesses potential to be molecules for model of IR-AML prognostic stratification. Genetic testing based on HTS contributes to revealing the pathogenic mechanism of AML, and is significant for evaluating the prognosis of patients with AML.


Subject(s)
High-Throughput Nucleotide Sequencing , Humans , Leukemia, Myeloid, Acute/genetics , Middle Aged , Mutation , Prognosis , Retrospective Studies
9.
Braz. j. med. biol. res ; 54(2): e9173, 2021. tab, graf
Article in English | LILACS, ColecionaSUS | ID: biblio-1142586

ABSTRACT

This study aimed to explore the correlation of kinesin family member 2A (KIF2A) expression with disease risk, clinical characteristics, and prognosis of acute myeloid leukemia (AML), and investigate the effect of KIF2A knockdown on AML cell activities in vitro. Bone marrow samples were collected from 176 AML patients and 40 healthy donors, and KIF2A expression was measured by real-time quantitative polymerase chain reaction. Treatment response, event-free survival (EFS), and overall survival (OS) were assessed in AML patients. In vitro, KIF2A expression in AML cell lines and CD34+ cells (from healthy donors) was measured, and the effect of KIF2A knockdown on AML cell proliferation and apoptosis in HL-60 and KG-1 cells was detected. KIF2A expression was greater in AML patients compared to healthy donors, and receiver operating characteristic curve indicated that KIF2A expression predicted increased AML risk (area under curve: 0.793 (95%CI: 0.724-0.826)). In AML patients, KIF2A expression positively correlated with white blood cells, monosomal karyotype, and high risk stratification. Furthermore, no correlation of KIF2A expression with complete remission or hematopoietic stem cell transplantation was found. Kaplan-Meier curves showed that KIF2A expression was negatively correlated with EFS and OS. In vitro experiments showed that KIF2A was overexpressed in AML cell lines (KG-1, HL-60, ME-1, and HT-93) compared to CD34+ cells, moreover, cell proliferation was reduced but apoptosis was increased by KIF2A knockdown in HL-60 and KG-1 cells. In conclusion, KIF2A showed potential to be a biomarker and treatment target in AML.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Leukemia, Myeloid, Acute/diagnosis , Leukemia, Myeloid, Acute/genetics , Kinesin/genetics , Biomarkers, Tumor/genetics , Survival Rate , Risk Factors , Apoptosis , HL-60 Cells , Cell Proliferation , Gene Knockdown Techniques
10.
Braz. j. med. biol. res ; 54(2): e9869, 2021. tab, graf
Article in English | LILACS, ColecionaSUS | ID: biblio-1142585

ABSTRACT

Severe blockage in myeloid differentiation is the hallmark of acute myeloid leukemia (AML). Trdmt1 plays an important role in hematopoiesis. However, little is known about the function of Trdmt1 in AML cell differentiation. In the present study, Trdmt1 was up-regulated and miR-181a was down-regulated significantly during human leukemia HL-60 cell differentiation after TAT-CT3 fusion protein treatment. Accordingly, miR-181a overexpression in HL-60 cells inhibited granulocytic maturation. In addition, our "rescue" assay demonstrated that Trdmt1 3′-untranslated region promoted myeloid differentiation of HL-60 cells by sequestering miR-181a and up-regulating C/EBPα (a critical factor for normal myelopoiesis) via its competing endogenous RNA (ceRNA) activity on miR-181a. These findings revealed an unrecognized role of Trdmt1 as a potential ceRNA for therapeutic targets in AML.


Subject(s)
Humans , Leukemia, Myeloid, Acute/genetics , MicroRNAs/genetics , DNA (Cytosine-5-)-Methyltransferases/genetics , Cell Differentiation , HL-60 Cells
11.
Rev. cuba. hematol. inmunol. hemoter ; 36(3): e1164, jul.-set. 2020. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1156440

ABSTRACT

Introducción: En el Instituto de Hematología e Inmunología se realiza el estudio molecular de las leucemias mieloides agudas (LMA). Para las leucemias mieloides agudas no promielocíticas (LPM) se determinan cuatro biomarcadores: los genes de fusión RUNX1-RUNX1T1 y CBF(-MYH11, la duplicación interna en tándem del gen FLT3 (DIT FLT3) y la mutación A del gen NPM1 (NPM1-A). Objetivo: Determinar la frecuencia de estos cuatro biomarcadores, en pacientes cubanos con leucemias mieloides agudas primaria no promielocíticas. Métodos: Se incluyeron 91 pacientes entre niños y adultos, estudiados en el Instituto durante tres años desde el debut. A partir de ARN de sangre medular se obtuvo ADN complementario por transcripción inversa; se amplificaron los fragmentos correspondientes mediante la reacción en cadena de la polimerasa y el producto se analizó por electroforesis capilar. Resultados: El RUNX1-RUNX1T1 apareció en el 24,2 por ciento, fue más frecuente en los pacientes pediátricos y disminuyó significativamente con la edad. El CBFβ-MYH11 solo se encontró en adultos (4,8 por ciento). La NPM1-A con 41 por ciento fue mayoritaria entre los adultos. La DIT FLT3 se observó en el 21,6 por ciento y no mostró relación con la edad. NPM1-A y DIT FLT3 fueron las aberraciones con mayor presencia simultánea. Conclusiones: Por primera vez se describe la frecuencia de los cuatro biomarcadores moleculares en los pacientes cubanos con leucemias mieloides agudas primaria no promielocíticas; su comportamiento fue similar a lo descrito por otros autores, aunque se encontraron algunas particularidades(AU)


Introduction: At the Institute of Hematology and Immunology, the molecular study of acute myeloid leukemias (AML) is carried out. For nonpromyelocytic acute myeloid leukemias, four biomarkers are determined: the RUNX1-RUNX1T1 and CBF(-MYH11 fusion genes, the internal tandem duplication of the FLT3 gene (DIT FLT3), and the A mutation of the NPM1 gene (NPM1-A). Objective: To determine the frequency of these four biomarkers in Cuban patients with nonpromyelocytic primary acute myeloid leukemias. Methods: 91 patients were included, children and adults, who were studied at the Institute for three years from their disease debut. Complementary DNA was obtained from medullary blood RNA by reverse transcription. The corresponding fragments were amplified by polymerase chain reaction and the product was analyzed by capillary electrophoresis. Results: RUNX1-RUNX1T1 appeared in 24.2 percent; it was more frequent in pediatric patients and decreased significantly with age. CBFβ-MYH11 was found only in adults (4.8 percent). NPM1-A, accounting for 41 percent, represented the majority among adults. FLT3 DIT was observed in 21.6 por ciento and was not related to age. NPM1-A and DIT FLT3 were the disorders with the greatest concurrence. Conclusions: For the first time, the frequency of the four molecular biomarkers is described in Cuban patients with primary non-promyelocytic acute myeloid leukemias. Its characterization was similar to that described by other authors, although some peculiarities were found(AU)


Subject(s)
Humans , Biomarkers , Leukemia, Myeloid, Acute/genetics , Polymerase Chain Reaction , DNA, Complementary , Reverse Transcription , Electrophoresis, Capillary
12.
Article in Chinese | WPRIM | ID: wpr-880787

ABSTRACT

OBJECTIVE@#To explore the rate and distribution of Runt- related transcription factor 1 (RUNX1) gene mutations in patients with acute myeloid leukemia (AML) and the correlation of these mutations with the clinical characteristics and survival outcomes of the patients.@*METHODS@#The genomic DNA extracted from the bone marrow of 158 patients with newly diagnosed AML for PCR amplification of RUNX1 gene and sequence analysis to identify the mutations. The mutations of ASXL1, DNMT3A, TET2, FLT3, CEBPA, NPM1, IDH2, NRAS and c-KIT genes were also examined to analyze their association with RUNX1 gene mutations.@*RESULTS@#Among the 158 AML patients, 19 (12.0%) were found to have RUNX1 mutations in A166G (9 cases), A142T (6 cases) and A162L (4 cases). RUNX1 mutations were more frequent in elderly patients (@*CONCLUSIONS@#RUNX1 gene mutations are associated with an adverse prognosis of patients with AML.


Subject(s)
Core Binding Factor Alpha 2 Subunit/genetics , Humans , Leukemia, Myeloid, Acute/genetics , Mutation , Prognosis , Remission Induction
13.
Journal of Experimental Hematology ; (6): 1985-1990, 2020.
Article in Chinese | WPRIM | ID: wpr-880003

ABSTRACT

OBJECTIVE@#To investigate the clinical significance of the targeted next-generation sequencing assay for patients with suspected myeloid malignancies.@*METHODS@#A total of 39 hematopenia patients with suspected myeloid malignamies in Department of Hematology of The Affiliated Huai'an No.1 People's Hospital of Nanjing Medical University from January 2018 to April 2019 were treated, 20 hot spot genes of myelodysplastic syndrome (MDS) were detected.@*RESULTS@#Regarding the diagnostic type, there were 7 cases of idiopathic cytopenia of undetermined significance (ICUS), 8 cases of clonal cytopenias of undetermined significance (CCUS) and 24 cases of myeloid myeloid malignancies which included 18 cases of MDS, 4 cases of myelodysplastic/myeloproliferative neoplasms (MDS/MPN) and 2 cases of acute myeloid leukemia. Positive mutation was detected in 70.8% (17/24) of myeloid malignancy patients , and 72.7% (16/22) in MDS and MDS/MPN patients. The main mutation types were ASXL1, TET2 and RUNX1. Compared with gene negative group, there were no significant differences in sex, age (<60 years old or ≥60 years old), proportion of bone marrow blast cells (<5% or≥5%) and cytogenetics (good, medium and poor) (P>0.05). Furthermore, all 8 CCUS patients showed positive mutation, and the incidence of double or multiple mutation in CCUS group was significantly lower than that of the MDS and MDS/MPN group (37.5% vs 54.5%) (P=0.002). The mutation types between the two groups were similar, and there was no significant difference in variant allele frequency (P>0.05).@*CONCLUSION@#Our results suggest that there are high rates of double or multiple mutations in myeloid malignancies, especially in patients with MDS and MDS/MPN. Targeted sequencing assay can improve the diagnosis of myeloid malignancies, and guide clinical treatment.


Subject(s)
Humans , Leukemia, Myeloid, Acute/genetics , Middle Aged , Mutation , Myelodysplastic Syndromes/genetics , Myelodysplastic-Myeloproliferative Diseases , Patients
14.
Journal of Experimental Hematology ; (6): 1880-1884, 2020.
Article in Chinese | WPRIM | ID: wpr-879987

ABSTRACT

OBJECTIVE@#To investigate the expression of CD123 in patients with acute myeloid leukemia (AML) and its relationship between clinical features, concomitant fusion gene or gene mutation, efficacy and prognosis.@*METHODS@#365 patients with newly diagnosed AML (except M3) treated in the First Affiliated Hospital of Zhengzhou University were enrolled and retrospective analysis, and multi-parameter flow cytometry was performed to detect the expression of CD123 in myeloid leukemia cell population. CD123≥20% was defined as positive. Clinical features, concomitant fusion gene or gene mutation, efficacy and prognosis of CD123@*RESULTS@#The positive rate of CD123 in 365 newly diagnosed AML patients was 38.9%. Compared with the CD123@*CONCLUSION@#CD123 positive indicates that AML patients have higher tumor burden and are more difficult to reach remission. It is an independent risk factor for OS and EFS in patients with normal karyotype and intermediate risk, which is important to evaluate the prognosis of patients with AML without specific prognostic marker.


Subject(s)
Humans , Interleukin-3 Receptor alpha Subunit , Karyotype , Leukemia, Myeloid, Acute/genetics , Mutation , Patients , Prognosis , Retrospective Studies
15.
Journal of Experimental Hematology ; (6): 1853-1858, 2020.
Article in Chinese | WPRIM | ID: wpr-879983

ABSTRACT

OBJECTIVE@#To investigate the effect of miR-29b-3p on apoptosis and proliferation of acute myeloid leukemia (AML) cells by targeting signal transducer and activator of transcription 3 (STAT3).@*METHODS@#TargetScan and miRanda online databases were used to predict the binding sites of miR-29b-3p and STAT3 3'UTR. The targeting relationship between them was estimated by Dual-Luciferase reporter assay experiment. After miR-29b-3p over-expression, qPCR and Western blot were used to detect the expression of STAT3 mRNA and proteins, flow cytometry to determine the apoptosis of AML cells, and MTS to detect the changes of cell proliferation in each group.@*RESULTS@#Dual-Luciferase reporter assay confirmed that STAT3 was the target gene of miR-29b-3p. After miR-29b-3p overexpression, the expression of STAT3 mRNA and protein decreased. Compared with the control groups, the proliferation of AML cells in the overexpression group decreased and the apoptosis increased (P<0.05).@*CONCLUSION@#MiR-29b-3p can inhibit the proliferation and induce apoptosis of AML cells by down-regulating STAT3.


Subject(s)
Apoptosis , Cell Proliferation , Humans , Leukemia, Myeloid, Acute/genetics , MicroRNAs/genetics , STAT3 Transcription Factor/genetics
16.
Chinese Medical Journal ; (24): 2829-2839, 2020.
Article in English | WPRIM | ID: wpr-877939

ABSTRACT

BACKGROUND@#Acute myeloid leukemia (AML) is a malignant hematological disease, originating from hematopoiesis stem cell differentiation obstruction and clonal proliferation. New reagents or biologicals for the treatment of AML are urgently needed, and exosomes have been identified as candidate biomarkers for disease diagnosis and prognosis. This study aimed to investigate the effects of exosomes from bone marrow mesenchymal stem cells (BMSCs) on AML cells as well as the underlying microRNA (miRNA)-mediated mechanisms.@*METHODS@#Exosomes were isolated using a precipitation method, followed by validation using marker protein expression and nanoparticle tracking analysis. Differentially expressed miRNAs were identified by deep RNA sequencing and confirmed by quantitative real-time polymerase chain reaction (qPCR). Cell proliferation was assessed by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt method, and cell cycle progression and apoptosis were detected by flow cytometry. Functional gene expression was analyzed by qPCR and Western blotting (WB). Significant differences were determined using Student's t test or analysis of variance.@*RESULTS@#BMSCs-derived exosomes effectively suppressed cell proliferation (both P < 0.0001 at 10 and 20 μg/mL) and cell cycle progression (P < 0.01 at G0-G1 stage), and also significantly enhanced cell apoptosis (P < 0.001) in KG-1a cells. There were 1167 differentially expressed miRNAs obtained from BMSCs-derived exosomes compared with KG-1a cell-derived exosomes (P < 0.05). Knockdown of hsa-miR-124-5p in BMSCs abrogated the effects of BMSCs-derived exosomes in regulating KG-1a such as the change in cell proliferation (both P < 0.0001 vs. normal KG-1a cell [NC] at 48 and 72 h). KG-1a cells treated with BMSCs-derived exosomes suppressed expression of structural maintenance of chromosomes 4 (P < 0.001 vs. NC by qPCR and P < 0.0001 vs. NC by WB), which is associated with the progression of various cancers. This BMSCs-derived exosomes effect was significantly reversed with knockdown of hsa-miR-124-5p (P < 0.0001 vs. NC by WB).@*CONCLUSIONS@#BMSCs-derived exosomes suppress cell proliferation and cycle progression and promote cell apoptosis in KG-1a cells, likely acting through hsa-miR-124-5p. Our study establishes a basis for a BMSCs-derived exosomes-based AML treatment.


Subject(s)
Apoptosis/genetics , Cell Proliferation/genetics , Exosomes/genetics , Humans , Leukemia, Myeloid, Acute/genetics , Mesenchymal Stem Cells , MicroRNAs/genetics
17.
Braz. j. med. biol. res ; 52(6): e8424, 2019. tab, graf
Article in English | LILACS | ID: biblio-1001535

ABSTRACT

Although rare, CALM/AF10 is a chromosomal rearrangement found in immature T-cell acute lymphoblastic leukemia (T-ALL), acute myeloid leukemia, and mixed phenotype acute leukemia of T/myeloid lineages with poor prognosis. Moreover, this translocation is detected in 50% of T-ALL patients with gamma/delta T cell receptor rearrangement, frequently associated with low expression of transcription factor CCAAT/enhancer-binding protein alpha (CEBPA). However, the relevance of CEBPA low expression for CALM/AF10 leukemogenesis has not yet been evaluated. We generated double mutant mice, which express the Lck-CALM/AF10 fusion gene and are haploinsufficient for the Cebpa gene. To characterize the hematopoiesis, we quantified hematopoietic stem cells, myeloid progenitor cells, megakaryocyte-erythrocyte progenitor cells, common myeloid progenitor cells, and granulocyte-macrophage progenitor cells. No significant difference was detected in any of the progenitor subsets. Finally, we tested if Cebpa haploinsufficiency would lead to the expansion of Mac-1+/B220+/c-Kit+ cells proposed as the CALM/AF10 leukemic progenitor. Less than 1% of bone marrow cells expressed Mac-1, B220, and c-Kit with no significant difference between groups. Our results showed that the reduction of Cebpa gene expression in Lck-CALM/AF10 mice did not affect their hematopoiesis or induce leukemia. Our data corroborated previous studies suggesting that the CALM/AF10 leukemia-initiating cells are early progenitors with lymphoid/myeloid differentiating potential.


Subject(s)
Animals , Rabbits , Leukemia, Myeloid, Acute/genetics , CCAAT-Enhancer-Binding Protein-alpha/genetics , Haploinsufficiency/genetics , Hematopoiesis/genetics , Phenotype , Transcription Factors/genetics , Translocation, Genetic/genetics , Mice, Transgenic , Acute Disease , Flow Cytometry , Genotype
18.
Säo Paulo med. j ; 136(4): 361-367, July-Aug. 2018. tab, graf
Article in English | LILACS | ID: biblio-962733

ABSTRACT

ABSTRACT CONTEXT: Complex karyotypes in acute myeloid leukemia (AML) are characterized by an overall low response rate with frequent relapses after clinical treatment. CASE REPORT: Here, we describe the case of a 61-year-old obese female with clinically diagnosed AML who presented a complex karyotype involving an uncommon abnormality: ring chromosome 11. Immunophenotypic analysis confirmed the diagnosis. Classical and molecular cytogenetic analyses, using GTG banding and FISH (fluorescence in situ hybridization), revealed the presence of complex structural rearrangement involving r(11), add(12)(p13), der(5) and der(13). CONCLUSIONS: Molecular cytogenetic analysis is suitable for better identification and characterization of chromosomal rearrangements in AML. Case reports like this, as well as population-based studies, are necessary for understanding the karyotypic changes that occur in humans.


Subject(s)
Humans , Female , Middle Aged , Ring Chromosomes , Leukemia, Myeloid, Acute/genetics , Translocation, Genetic , In Situ Hybridization, Fluorescence , Cytogenetic Analysis , Karyotype
19.
Einstein (Säo Paulo) ; 15(3): 355-358, July-Sept. 2017.
Article in English | LILACS | ID: biblio-891404

ABSTRACT

ABSTRACT Acute myeloid leukemia is a hematopoietic stem cell neoplastic disease associated with high morbidity and mortality. The presence of FLT3 internal tandem duplication mutations leads to high rates of relapse and decreased overall survival. Patients with FLT3 internal tandem duplication are normally treated with hematopoietic stem cell transplantation in first complete remission. Nevertheless, the incidence of post-transplant relapse is considerable in this group of patients, and the management of this clinical condition is challenging. The report describes the outcomes of patients with FLT3 internal tandem duplication positive acute myeloid leukemia who relapsed after allogeneic hematopoietic stem cell transplantation and were treated with the combination of re-induction chemotherapy, donor lymphocyte infusion, sorafenib and azacitidine. Three cases are described and all patients achieved prolonged complete remission with the combined therapy. The combination of induction chemotherapy followed by donor lymphocyte infusion, and the maintenance with azacitidine and sorafenib can be effective approaches in the treatment of post-hematopoietic stem cell transplant and relapsed FLT3 internal tandem duplication positive acute myeloid leukemia patients. This strategy should be further explored in the context of clinical trials.


RESUMO A leucemia mieloide aguda é uma doença neoplásica de células-tronco hematopoiéticas com alta morbimortalidade. A presença de mutações de duplicação em tandem de FLT3 leva a altas taxas de recorrência e a menor sobrevida global. Os pacientes com duplicação em tandem de FLT3 são normalmente tratados com transplante de células-tronco hematopoiéticas na primeira remissão completa. No entanto, a incidência de recidiva pós-transplante é considerável neste grupo de pacientes, e a conduta, nestes casos, é um desafio. O relato descreve os resultados do tratamento de pacientes com leucemia mieloide aguda positiva e duplicação em tandem de FLT3 que recidivaram depois do transplante alogênico de células-tronco hematopoiéticas e que foram tratados com combinação de quimioterapia de reindução, infusão de linfócitos de doador, sorafenib e azacitidina. São descritos três casos, e todos os pacientes apresentaram remissão completa prolongada com a terapia combinada. A combinação de quimioterapia de indução, seguida de infusão de linfócitos do doador, e a manutenção com azacitidina e sorafenib podem ser abordagens eficazes no tratamento da recorrência pós-transplante em pacientes com leucemia mieloide aguda e duplicação em tandem de FLT3. Essa estratégia deve ser mais explorada no contexto de ensaios clínicos.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Phenylurea Compounds/administration & dosage , Azacitidine/administration & dosage , Leukemia, Myeloid, Acute/therapy , Niacinamide/analogs & derivatives , Lymphocyte Transfusion , fms-Like Tyrosine Kinase 3/genetics , Induction Chemotherapy , Antineoplastic Agents/administration & dosage , Recurrence , Leukemia, Myeloid, Acute/genetics , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Treatment Outcome , Niacinamide/administration & dosage , Combined Modality Therapy/methods , Neoplasm Recurrence, Local/therapy
20.
Rev. AMRIGS ; 61(1): 19-24, jan.-mar. 2017. tab, graf
Article in Portuguese | LILACS | ID: biblio-849061

ABSTRACT

Introdução: Análise do perfil citogenético de pacientes com Leucemia mieloide aguda (LMA) e correlação com dados clínico-epidemiológicos e seus respectivos prognósticos. Métodos: Busca de registros nos prontuários de pacientes com diagnóstico de LMA no período de janeiro de 2009 a dezembro de 2014. Mensurou-se o perfil citogenético da amostra, estratificando-a por faixa etária e classificação conforme FAB e OMS para posterior correlação dos dados de prognóstico e distribuição epidemiológica obtidos com os dados disponíveis na literatura médica atual. Resultados: Foram selecionados 38 pacientes para análise, com idades entre 1 e 84 anos. Todos os pacientes possuíam cariotipagem e 76,3% apresentavam alterações citogenéticas; 13 pacientes apresentavam cariótipo favorável (CF), 14, cariótipo intermediário (CI), 10, cariótipo desfavorável (CD) e 1 apresentava LMA secundária. No grupo com CF, a média de idade foi de 25 anos (p=0,0151); no grupo com CD, foi de 39,8%. No grupo com CF, 69,2% obtiveram remissão completa com protocolo de tratamento; no grupo com CI, 18,18% obtiveram a remissão completa, e por fim, no grupo com CD nenhum caso obteve remissão completa. Houve tendência à plaquetopenia mais significativa no grupo com CI. Conclusão: Ficou nítida a correlação das alterações citogenéticas observadas com sua respectiva importância prognóstica, corroborando a necessidade da determinação do cariótipo para o adequado manejo dos pacientes com LMA (AU)


Introduction: An analysis of the cytogenetic profile of patients with acute myeloid leukemia (AML) and correlation with clinical and epidemiological data and their respective prognoses. Methods: Search of records in the medical charts of patients diagnosed with AML from January 2009 to December 2014. The cytogenetic profile of the sample was measured by stratifying it by age group and classification according to FAB and WHO for subsequent correlation of prognostic and epidemiological distribution data obtained from the available data of the medical literature. Results: Thirty-eight patients were selected for analysis, ranging from 1 to 84 years of age. All patients had karyotyping and 76.3% presented cytogenetic alterations; 13 patients had favorable karyotype (FK), 14 intermediate karyotype (IK), 10 unfavorable karyotype (UK), and 1 had secondary AML. In the FK group, the mean age was 25 years (p = 0.0151); in the UK group, 39.8%. In the FK group, 69.2% achieved complete remission with treatment protocol; in the IK group, 18.18% had complete remission, and finally, in the UK group, no case achieved complete remission. There was a trend towards more significant thrombocytopenia in the IK group. Conclusion: The correlation of the observed cytogenetic changes with their respective prognostic importance was clear, corroborating the need to determine the karyotype for an adequate management of patients with AML (AU)


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Leukemia, Myeloid, Acute/epidemiology , Leukemia, Myeloid, Acute/genetics , Brazil/epidemiology , Chromosome Aberrations , Cross-Sectional Studies , Cytogenetic Analysis
SELECTION OF CITATIONS
SEARCH DETAIL