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ABSTRACT Acute erythroid leukemia (AEL) is an exceedingly uncommon but distinct hematological malignancy that shows neoplastic proliferation of erythroid precursors with maturation arrest and no significant myeloblasts. We describe an autopsy case of this rare entity in a 62-year-old man with co-morbidities. He underwent a bone marrow (BM) examination for pancytopenia during the first outpatient department visit, which revealed an increased number of erythroid precursors with dysmegakaryopoiesis suggesting the possibility of Myelodysplastic syndromes (MDS). Thereafter, his cytopenia got worsened, warranting blood and platelet transfusions. Four weeks later on the second BM examination, AEL was diagnosed based on morphology and immunophenotyping. Targeted resequencing for myeloid mutations revealed TP53 and DNMT3A mutations. He was initially managed along febrile neutropenia with the stepwise escalation of antibiotics. He developed hypoxia attributed to anemic heart failure. Subsequently, he had hypotension and respiratory fatigue pre-terminally and succumbed to his Illness. A complete autopsy showed infiltration of various organs by AEL and leukostasis. Besides, there was extramedullary hematopoiesis, arterionephrosclerosis, diabetic nephropathy (ISN-RPS class II), mixed dust pneumoconiosis, and pulmonary arteriopathy. The histomorphology of AEL was challenging, and the differential diagnoses were many. Thus, this case highlights the autopsy pathology of AEL, an uncommon entity with a strict definition, and its relevant differentials.
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Objective@#To investigate the synergistic lethal effect and mechanism of arsenic trioxide (ATO) and aclacinomycin (ACM) on human acute myeloid leukemia cell line KG-1a.@*Methods@#Colony-forming assay was used to detect the proliferation of KG-1a cells treated with different concentration of ATO and ACM. Compusyn software was used to analyze the synergistic effect of ATO and ACM. Flow cytometry and Wright's staining were used to analyze the apoptotic rate of KG-1a cells induced by combined treatment of ATO and ACM. Western blot was used to determine the expression of proteins associated with apoptosis.@*Results@#The cytotoxicity of arsenic trioxide or aclacinomycin alone was in a dose-dependent manner. Flow cytometry analysis showed that the apoptotic rate of KG-1a cells treated with both 0.4 μmol/L ATO and 10 nmol/L ACM was (34.5±3.1)%, significantly higher than (7.6±1.1)% of 0.4 μmol/L ATO treatment or (18.7±2.3) % of 10 nmol/L ACM treatment alone (P<0.05). The apoptotic rate of KG-1a cells treated with both 1.5 μmol/L ATO and 37.5 nmol/L ACM was (52.5±4.7)%, significantly higher than (19.1±3.2)% of 1.5 μmol/L ATO treatment or (27.7±2.2)% of 37.5 nmol/L ACM treatment alone (P<0.05). The apoptotic rate of KG-1a cells treated with both 3.0 μmol/L ATO and 75 nmol/L ACM was (61.3±4.5)%, significantly higher than (29.5±2.5)% of 3.0 μmol/L ATO treatment or (28.6±3.4) % of 75 nmol/L ACM treatment alone (P<0.05). In addition, the result of Wright's staining showed that combined treatment of ATO and ACM induced a more apparent phenotype of apoptosis when compared with single agent treatment. Compusyn software analysis showed that the combination index (CI) value of combined treatment group was less than 1, which indicated the synergistic effect of these two agents.@*Conclusions@#Combined treatment of ATO and ACM shows a synergistic lethal effect on human acute myeloid leukemia cell line KG-1a via activating the apoptotic pathway, which inhibits cell growth and induces apoptosis.
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Objective To observe the biological characteristics and analyse primary therapeutic response of acute erythroid leukemia.Methods The data of 28 patients primarily diagnosed as acute erythroid leukemia were analyzed.The patients were divided into with muhilineage dysplasia group and without muhilineage dysplasia group,and the morphology,immunology,cytogenetics and molecular biology characteristics and the complete remission rate of the first induction therapy were compared.Results There were 14 cases(50％)with muhilineage dysplasia,which involved in two lineage or trilineage.In 6 cases by flow cytometry,the myeloid blast immunophenotypes were common expressed.In 8 cases detected by karyotype analysis,5 cases were chromosomal abnormal,including 4 cases were complex chromosomal abnormal,1 case was trisomy 8.In 4 cases underwent WT1 detection,all of them were positive.The complete remission rate of the first induction therapy was 39.29％(11/28),the ratein the multilineage dysplasia group was 35.71％(5/14),and the ratein without multilineage dysplasia group was 42.86％(6/14),the difference had no statistical significance(P＞0.05).The complete remission rate of the complex chromosome group was 25.00％(1/4),the intermediate prognostic group was 50.00％(2/4).Conclusions Acute erythroid leukemia had special biological features different from other subtype AML:accompanyed with high frequency of multilineage dysplasia.The abnormality of karyotype were high,and it was often complex karyotype involved with chromosome 5 and/or chromosome 7,which had a low complete remission rate.The complete remission rate of chemotherapy was low,treatment effect was poor.
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Objective To investigate the biological characteristics of mouse erythrolenkemia cell FBL-3.Methods The morphological feature, growth characters, clone formation and immunochemistry of mouse erythroleukemia cell FBL-3 were examined by light microscope. The cell cycle distribution and expression of MHC were detected by flow cytometry. Drug sensitivity was measured by MTT assay. Tumorigenicity was evaluated after intravenous injection FBL-3 cells into C57BL/6 mice. Results FBL-3 cells were smaller,fusiform or polygon, adherence. Doubling time was 24.12 h. The clone formation rates were (35.23±1.44) %and (60.27±5.56) % at 14 th and 21 th day, respectively. The reactions for PAS and chloracetic acid were positive, while the POX, NAP and butanoic acid reactions were negative. The cell cycle distribution was as follow: G0/G1 phase (50.9±2.5) %, S phase (36.3±1.4) %, G2/M phase (13.8±0.8) %. The IC50 of FBL-3 cells to Ara-C, VDS, DDP, MMC and MTX were (0.49±0.04), (0.87±0. 09), (3.77±0.32), (1.66±0.16) μmol/L and (2.77±0.24) nmol/L, respectively. Chromosome number was at 34 to 41. MHC of FBL-3 cell was H-2b. Sexual gene Sry was positive. All C57BL/6 mice were morbidity with erythroblastic leukemia when FBL-3 cells had been intravenously inoculated. There was a linear relationship between the survival time and the number cell injected. The main targets for the leukemic FBL-3 cells were liver, spleen, kindey and lung. Conclusion FBL-3 cell has typical features of mouse leukemia cell, was easily cultured in vitro and tumorigenesised in C57BL/6 mice. FBL-3 cell could be as a satisfactory tool for the research of leukemia.
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AIM: To clone human ?-globin gene carrying a thalassemic mutation IVS II654(C→T) and establish a eukaryotic expression system for high-level expression of human ? IVS II654 gene in mouse erythroleukaemia(MEL) cells. METHODS: The fragments of human ? 654 gene isolated from the ? thalassemia patients homozygous for the ? 654 mutation were amplified by PCR, and cloned to plasmid pBGT51. Then, the human ? LCR and ? 654 gene were subcloned from plasmid pBGT51 to the stable mammalian expression vector pcDNA3.1+ together, and the MEL cells were transfected with this vector using commercially available cationic lipid FuGENE6. The MEL cells were induced for further maturation by DMSO and the expression of human ? 654 gene in the MEL cells was identified by RT-PCR. RESULTS: A mammalian expression system of human ? thalassemic mutation ?IVS II654(C→T) was established. CONCLUSION: The level and the reliability of expression of human ? 654 gene in the MEL cells in vitro are similar to that in vivo in human body. This may be a valuable gene therapy model for human ? thalassemic mutation ?IVS II654(C→T).
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Objective To investigate the photodynamic therapy(PDT)of human erythroleukemia cell line K562 and drug-resistance cell line K562/ADM using hexyl 5-aminolevulinate(He-ALA)in vitro.Methods Four groups were set:PDT group(photosensitizer with light irradiation),laser group(light irradiation only),dark-cytoxocity group(photosensitizer only)and normal control group(neither photosensitizer nor light irradiation).K562 cells and K562/ADM cells cultured were incubated with various concentrations of He-ALA(0.025,0.1,0.4 and 1.6mmol/L)for 4 hours,then illuminated with different light doses(4.5,9,18 and 36 J/cm2)using a laser with a wave length of 630nm.After 12 hours incubation,Wright's staining method was used to observe the cell's morphological changes,the survival rates of cells were analyzed by CCK8 assay,and the changes in colony-forming abilities of cells were analyzed by methylcellulose colony-forming assay.Results He-ALA or light irradiation alone had no evident cytotoxicity,while the application of He-ALA plus light irradiation effectively killed the leukemia cells.With the increase of the concentrations of He-ALA and the dosages of irradiation,the viability of cells decreased.Under the same conditions,the survival rate of K562 was significantly lower than that of K562/ADM(P