The study was purposed to investigate the expression of CD73 on bone marrow nucleated cells (BMMNC) in various leukemia subtypes and its relationship with cell differentiation of leukemia. Immunocytochemistry staining and Wright-Giemsa staining of BMMNC from 75 cases of leukemia, 11 cases of myelodysplastic syndrome (MDS), 13 cases of non-leukemic patients and 9 healthy adults were performed, and the CD73(+) ratio in BMMNC and its relationship with differentiation of leukemia cells were analyzed. The results showed that the ratios of CD73(+) in BMMNC of com-B ALL, pre-B ALL and PLL were significantly higher than those in B-CLL (p < 0.05). CD73(+) ratios in AML subtypes of M(1), M(2a), t (8; 21), t (15; 17), M(4) and M(5) were markedly higher than those in MDS respectively, but in M(6) and MDS were lower and had no statistical difference between them. CD73(+) ratios in T-ALL, B-CLL, M(6), MDS, non-leukemia patients and healthy adults were close to each other and all of them were lower than those in B-ALL and other AML subtypes. It is concluded that the expression of CD73 is associated with leukemia subtype, differentiation and development. The higher differentiation of leukemia cells, the lower of CD73 expression in myeloid and B lymphoid leukemia, but T-ALL does not meet this pattern.
5'-Nucleotidase ; metabolism ; Adolescent ; Adult ; Cell Differentiation ; Humans ; Leukemia ; metabolism ; pathology ; Leukemia, Lymphocytic, Chronic, B-Cell ; metabolism ; Leukemia, Myeloid, Acute ; metabolism ; Myelodysplastic Syndromes ; metabolism ; Young Adult

Hypercalcemia in accelerated phase of chronic myelogenous leukemia (CML) is very rare. Its pathogenesis is considered humoral hypercalcemia of malignancies mediated by parathyroid hormone-related protein (PTHrP). In severe hypercalcemia, calcifications in kidneys, skin, vessels, heart, and stomach may occur. Our two cases were admitted because of severe hypercalcemia in accelerated phase of CML. On Tc-99m methylene diphosphonate (MDP) bone scintigraphies, a marked tracer accumulation was seen in the lung, heart, stomach and kidney. We report increased tracer accumulation of multiple organs on Tc-99m MDP bone scintigraphy in two rare hypercalcemic patients with CML.
Adult ; Bone Diseases/radionuclide imaging* ; Bone Diseases/etiology* ; Calcinosis/radionuclide imaging ; Calcinosis/etiology ; Case Report ; Human ; Hypercalcemia/radionuclide imaging* ; Hypercalcemia/etiology* ; Leukemia, Myeloid, Chronic/metabolism ; Leukemia, Myeloid, Chronic/complications* ; Male ; Middle Age ; Proteins/metabolism ; Technetium/diagnostic use

To explore the effect of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (ASODN) on telomerase activity in primary acute myeloid leukemia (AML) cells and chronic myeloid leukemia (CML) cells, polymerase chain reaction enzyme-linked immunosorbent assay (PCR-ELISA) was used to determine telomerase activity. The expression levels of hTERT protein were assayed by flow cytometry using immunofluorescence labeling. Immunofluorescence assay showed that the expression levels of hTERT protein in AML and CML cells decreased with time after hTERT ASODN treatment. There was no difference in hTERT protein levels between control and sense oligodeoxynucleotide (SODN)-treated cells. Telomerase activity decreased when AML and CML cells were treated with ASODN for 48 h. Telomerase activity of AML and CML cells was significantly inhibited when the cells treated with ASODN for 72 h. There was no difference in telomerase activity levels between control and hTERT SODN-treated cells. It was concluded that hTERT ASODN could inhibit hTERT protein expression level, thus decreasing the telomerase activity in primary AML and CML cells.
Adolescent ; Adult ; DNA-Binding Proteins ; Female ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; enzymology ; Leukemia, Myeloid, Acute ; enzymology ; Male ; Middle Aged ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Telomerase ; antagonists & inhibitors ; metabolism

This study was aimed to compare the expression level of eIF4E in patients with leukemia and normal controls, and to explore its role in leukemogenesis. White blood cells were collected in 76 leukemia patients and 10 healthy volunteers. The mRNA and protein expressions of eIF4E were detected by QT-PCR and Western blot in 39 cases of acute myeloid leukemia (AML), 15 cases of chronic myeloid leukemia (CML), 22 cases of acute lymphocytic leukemia (ALL) and 10 healthy volunteers as normal controls. The results demonstrated that compared with normal controls, the absolute expression levels of eIF4E mRNA increased in patients with AML, ALL and CML in blastic phase (P < 0.05), but had no significant change between groups of CML in chronic and accelerated phase although some increasing in group of CML in accelerated phase. The relative expression level of eIF4E mRNA had no significant change in AML, ALL, CML groups except the two subtypes of leukemia M4 and M5. Furthermore, the protein expression level in group of CML in accelerated phase and blastic phase and all acute leukemia patients including AML and ALL were higher than that in normal controls (P < 0.05). It is concluded that although its mRNA relative expressions had no significant change in most leukemia patients, the absolute expression level of eIF4E mRNA and its protein expression is up-regulated in most leukemia patients, which may play an important role in leukemogenesis, so the eIF4E may be a promising target for leukemia therapy and eIF4E-targeted therapy may be an option especially for the relapse and refractory leukemia.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Eukaryotic Initiation Factor-4E ; genetics ; metabolism ; Female ; Humans ; Leukemia ; genetics ; metabolism ; pathology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Leukemia, Myeloid, Acute ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; RNA, Messenger ; genetics ; Young Adult

BACKGROUND: Adenosine deaminase (ADA), an enzyme involved in purine metabolism, catalyzes the hydrolytic deamination of adenosine or 2-deoxyadenosine to inosine or 2-deoxyinosine. Human ADA consists of three molecular forms: ADA1, ADA1+CP, and ADA2. The two ADA isoenzymes represent two different gene products and have different tissue distributions, and their concentrations in serum appear to reflect different pathological conditions or physiological responses. Elevation of serum ADA activity has been described especially in leukemia and lymphoma. The purpose of this study was to evaluate the clinical utility of ADA isoenzyme determination in the diagnosis of leukemia. METHODS: We studied the activity of serum ADA and its isoenzyme in 44 leukemic patients. The study population consisted of 17 patients with acute lymphoblastic leukemia (ALL), 23 with acute myeloid leukemia (AML), and 4 with chronic myelogenous leukemia (CML). ADA isoenzyme was measured by erythro-9- (2-hydroxy-3-nonyl) adenine [EHNA] inhibitory assay using the Hitachi 7170 autoanalyzer. RESULTS: The rates of abnormally high total ADA activity were 100% for ALL, 60.8% for AML, and 50% for CML. In isoenzyme pattern, there was a clear difference between ALL and AML. High level of ADA1 activity was found in patients with ALL (P <0.01). The ADA1/ADA2 ratio was significantly higher (P <0.001) in ALL than AML. There was a correlation between ADA1 and absolute number of peripheral blasts in AML (r=0.840). CONCLUSIONS: It is concluded that the measurement of ADA isoenzyme may be a useful biochemical marker for leukemic diagnosis.
Adenine ; Adenosine Deaminase* ; Adenosine* ; Biomarkers ; Deamination ; Diagnosis ; Humans ; Inosine ; Isoenzymes ; Leukemia* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Leukemia, Myeloid, Acute ; Lymphoma ; Metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Tissue Distribution

The aim of this study was to investigate the expression status of the helicase antigen (HAGE) transcript and its clinical significance in patients with acute myeloid leukemia (AML) and chronic myeloid leukemia (CML). The expression of HAGE cDNA in bone marrow mononuclear cells from AML and CML patients was detected by using real-time quantitative PCR. The results indicated that overexpression of HAGE transcript (117.12% - 9842.70%, median 434.96%) was detected in 14.8% (11/74) AML patients. AML patients with HAGE cDNA expression were significantly older than those HAGE-negative patients (median 67 and 45 years, respectively, p = 0.001). HAGE cDNA expression was more frequently present among the patients with acute monoblastic leukemia (M(4) and M(5), 7 of 20, 35.0%), compared to the patients with acute non-monoblastic leukemia (M(1), M(2), M(3) and M(6), 4 of 54, 7.4%) (p = 0.007). 28.6% (8/28) cases with normal karyotypes showed HAGE cDNA overexpression, significantly higher than 7.5% (3 of 40) in those with chromosomal abnormalities (p = 0.041). Overexpression of HAGE transcript was found in 9 (34.6%) CML cases and more frequently observed at accelerated phase and blast crisis (4/4, 100%) than that at chronic phase (5/22, 22.7%) (p = 0.008). It is concluded that HAGE cDNA expression is relevant to specific subtypes of AML and to the progression of CML.
Blast Crisis ; Bone Marrow Cells ; metabolism ; DEAD-box RNA Helicases ; genetics ; metabolism ; DNA, Complementary ; Gene Expression ; Humans ; Karyotype ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics

In order to investigate the suppressor of cytokine signaling-1 (SOCS-1) expression in peripheral blood mononuclear cells (PBMNC) of patients with acute and chronic myeloid leukemia and analyze its clinical significance, RT-PCR method was used for detecting SOCS-1 mRNA expression in PBMNC of 50 newly diagnosed patients. The result showed that positive expressions of SOCS-1 were observed in 4 of 25 patients with AML (16.00%), in 11 of 25 patients with CML (44.00%) and none in 10 normal controls. The differences between patients with AML and normal controls, and between patients with CML and normal controls were statistically significant. In CML group, 2 out of 12 cases with non-progression (chronic phase), 9 of 13 cases with progression showed the positive expression, the difference between two subgroups was statistically significant. Those CML patients with SOCS-1 mRNA expression had poor response to IFN-alpha. When they transformed into accelerated phase, SOCS-1 mRNA expression was more persistently and frequently observed, and no response to IFN-alpha was observed. Most of them had very poor prognosis. It is concluded that the SOCS-1 mRNA can be detected in the PBMNC of the patients with acute and chronic myeloid leukemia. The SOCS-1 mRNA expression in the patients with CML is higher than that in patients with AML, and it is higher in accelerated phase and blast crisis significantly. This phenomenon is highly related to the reaction of IFN-alpha and prognosis.
Humans ; Interferon-alpha ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Suppressor of Cytokine Signaling 1 Protein ; Suppressor of Cytokine Signaling Proteins ; biosynthesis ; genetics

OBJECTIVETo explore the specificity of anti-phosphotyrosine monoclonal antibody PY20 in bcr-abl+ cells and its possible clinical applications.

METHODSBcr-abl cell lines( K562, MEG-01) and bcr-abl- cells lines( Jurkat, MCF-7 )were stained with PY20. Phosphotyrosine protein of K562 and MEG-01 cells was detected by flow cytometry before and after treatment with imatinib. Phosphotyrosine protein in bone marrow cells from 49 patients with chronic myeloid leukemia (CML), Ph+ acute lymphoblastic leukemia(Ph(+) -ALL), Ph- ALL, acute myeloid leukemia (AML-M1, M2, M3, M5, FAB classification), chronic lymphocytic leukemia (CML) and 3 normal donor. Positive cells over 5% of total cells was considered positive cases for phosphotyrosine protein. The level of tyrosine phosphorylation was determined by median fluorescence intensity (MFI).

RESULTSBcr-abl cell lines and marrow cells from 10 CML patients and 8 ALL patients were all PY20-positive, while bcr-abl- cell lines and marrow cells from 18 leukemia patients and 3 normal donor were all PY20-negative. MFI decreased remarkably after blocked by imatinib in K562 cells and MEG-01 cells. The positive cell percent of marrow cells from 10 newly diagnosed CML patients and 9 imatinib-sensitive CML patients was (54.20 +/- 19.82)% and (14.84 +/- 6.17)% (P < 0.05), while that of 2 cases of imatinib-resistant was 64.3% and 57.2%. There was significant difference of MFI between imatinib-resistant patients and imatinib-sensitive patients (99.42 +/- 4.87 vs 46.41 +/- 4.67, P < 0.01).

CONCLUSIONPY20 monoclonal antibody is highly specific for bcr-abl+ cells. It might be useful in rapid detection of bcr-abl+ cells and sensitivity to imatinib of CML patients.

Antibodies, Monoclonal ; analysis ; Bone Marrow Cells ; metabolism ; Flow Cytometry ; Fusion Proteins, bcr-abl ; analysis ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Leukemia, Myeloid, Acute ; metabolism ; Phosphotyrosine ; analysis ; immunology

This study was aimed to investigate the expression level of CIAPIN1 mRNA in leukemia patients and explore its significance in leukemias. The fresh bone marrow was collected from 112 newly diagnosed leukemia patients, the total RNA was extracted by means of TRIzoL, the cDNA was synthesized, the expression of CIAPIN1 mRNA was detected by real-time quantitative PCR using β-actin as internal reference; 10 normal healthy persons were selected as controls. The results showed that the expression of CIAPIN1 mRNA was statistically higher in acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL) and chronic phase chronic myeloid leukemia (CML) patients than that in normal persons (p < 0.05); but there was no statistical difference between chronic lymphocytic leukemia (CLL) and normal persons (p > 0.05). It is concluded that the CIAPIN1 gene higher expresses in MNC of newly diagnosed leukemia patients, up-regulation of CIAPIN1 expression may play an important role in pathogenesis of leukemia.
Adult ; Bone Marrow Cells ; metabolism ; Case-Control Studies ; Female ; Gene Expression Regulation, Leukemic ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Leukemia ; genetics ; metabolism ; pathology ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Young Adult

To evaluate the expression of cyclin dependent kinase inhibitor P27(Kip1) in leukemia and to investigate its clinical significance, the P27(Kip1) protein in bone marrow or peripheral blood samples from 82 cases of leukemia was measured by Western blot and enhanced chemoluminescence (ECL). The results showed that the expression of P27(Kip1) protein in ALL was higher than that in ANLL (P = 0.033) and also that in CML (P = 0.008). P27(Kip1) expression in CLL was higher than that in CML too (P = 0.017). In acute leukemia, the effective rate (CR and PR) of initial chemical therapy in the group of P27(Kip1) > 0.655 was higher than that in the group of P27(Kip1) < or = 0.655, P = 0.041. For ANLL and ALL patients, the survival time in the group of P27(Kip1) > 0.655 was longer than that in the group of P27(Kip1) < or = 0.655, P = 0.0065. There were similar statistical significance for ANLL and ALL patients, P = 0.0271 and P = 0.0266 respectively. There was a negative correlation between chromosomal abnormalities and P27(Kip1) expression in ALL patients (r = -0.775, P = 0.04). The expression of P27(Kip1) protein appeared nothing to do with sex, age, white blood cell number, blast cell number in peripheral blood, serum LDH or uric acid. In conclusion, the expression level of P27(Kip1) protein is in relation to the effect of initial chemical therapy and survival time, so that the lower P27(Kip1) expression may associated with poor prognosis in acute leukemia.
Adolescent ; Adult ; Aged ; Blotting, Western ; Cell Cycle Proteins ; analysis ; Child ; Child, Preschool ; Chromosome Aberrations ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; Leukemia, Lymphocytic, Chronic, B-Cell ; drug therapy ; genetics ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; drug therapy ; genetics ; metabolism ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; metabolism ; Survival Rate ; Tumor Suppressor Proteins ; analysis