Objective To investigate the levels of interleukin-15(IL-15) in children with acute leukemia and contribute to understand their function on acute leukemia about the process.Methods Every group of acute lymphocytic leukemia(ALL),acute nonlymphocytic leukemia(ANLL) or non-leukemia group had 20 children who did not receive any treatment. Peripheral blood of every group and 20 other healthy children were aspirated .The levels of IL-15 in serum was analyzed by enzyme linked immunosorbent assay (ELISA).Results The levels of IL-15 in ALL group,ANLL group, non-leukemia group and healthy children were (34.37?2.8) ng/L, (29.61?3.2 )ng/L, (117.54?3.9) ng/L, (122.23?4.2) ng/L. Compared with the levels of the healthy group or the non-leukemia group, the difference of ALL or ANLL was signicant in statistics(q=3.95,4.03 Pa0.05).Conclusion Detection of serum IL-15 levels may have clinical significance in judging the anti-tumor immune condition in children with leukemia.

Objective To investigate children′s myelodysplastic hematopoietic cells reaction to interleukin (IL)-15.Methods CD 34 + cells in bone marrow from 18 myelodysplast syndrome(MDS) patients were purified by an immunomagnetic beads sorting system. Apoptosis of hematopoietic precursors was assayed by propidium iodine staining and flow cytometric analysis.Results On 8th cultured day,when IL-15 concentration was between 0-100 ng/ml,it could suppress apoptosis of hematopoietic cells in MDS patients in a dose-and- time dependent manner. IL-15 in study group significanthy lower than that of control group.Conclusion IL-15 may partly suppress apoptosis of hematopoietic cells in MDS patients.

Objective To analyze the chemokine receptor CXCR4 expression in acute leukemic cells of children and its relationship with extramedullary infiltration.Methods The immunotypes of cases of acute leukemia in children and the expression of CXCR4 in marrow leukemic cells were studied by flow cytometry respectively. The relationship between CXCR4 expression and extramedullary infiltration of leukemic cells were analyzed by statistical method.Results The expression rates of CXCR4 in ALL children were higher than those in NALL children(P

The objective of study was to explore the role of vascular endothelial growth factor (VEGF) and its receptor-2 in pathogenesis of acute myeloid leukemia. The acute myeloid leukemia model was established on 20 mice with severe combined immunodeficiency (SCID) transplanted by HL-60 cells. The mice were divided into the normal control and test group randomly. The expression of VEGF was detected by enzyme linked immunosorbent assay (ELISA). The expression of VEGFR-2 mRNA was detected by RT-PCR. The results showed that the establishment of acute myeloid leukemia model was succeeded on all SCID mice by HL-60 cell transplantation. The expressions of VEGF and VEGFR-2 mRNAs could be determined on all mice. As compared with the normal control group, the expressions of VEGF and VEGFR-2 mRNAs in the test group significantly increased, but gradually increased during the course of disease. It is concluded that the abnormal expressions of VEGF and VEGFR-2 exist in mice with acute myeloid leukemia, which may be involved in the pathogenesis of AML.
Animals ; Bone Marrow ; pathology ; HL-60 Cells ; Humans ; Mice ; Mice, SCID ; Neoplasm Transplantation ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

To study the effect of cytochrome C on HL-60 cells in vitro and the mechanism of expression changes of relevant apoptotic genes, the inhibition rate of cytochrome C on HL-60 cells was detected by MTT, the morphology of HL-60 cells was observed by light microscopy and fluorescence microscopy, the changes of apoptosis rate and cell cycle were assayed by flow cytometry (FCM), DNA ladder was investigated on electrophoresis, the expression changes of bax and bcl-2 mRNA were examined by RT-PCR, when HL-60 cells were treated with different concentrations of cytochrome C for 24 hours. The results showed that the inhibition rate increased with increase of the cytochrome C concentration within 0 - 150 mg/L; when treated with 0 - 37.5 mg/L cytochtome C for 24 hours, the percentage of apoptotic HL-60 cells increased with the dose increasing, and the typical apoptotic cells and the apoptotic DNA ladder were observed. At the same time, within this range of concentration, the expression of bcl-2 mRNA decreased gradually and the expression of bax increased gradually. When the cytochrome C concentration was higher than 37.5 mg/L, the percentage of apoptotic HL-60 cells not increased, but decreased, while the cells necrosed. The above metioned results suggested that at certain range of concentration of cytochrome C, apoptosis or necrosis can be induced by cytochrome C, and cell cycle arrests at G(1) phase in HL-60 cells, the percentage of apoptotic cells and the changes of expression of bax and bcl-2 depend on the dose of cytochrome C. The mechanism that cytochtome C induced apoptosis in HL-60 cells may be related to the activation of bax and inhibition of bcl-2.
Apoptosis ; drug effects ; Cytochromes c ; pharmacology ; Dose-Response Relationship, Drug ; Flow Cytometry ; Gene Expression Regulation, Leukemic ; drug effects ; HL-60 Cells ; Humans ; Microscopy, Fluorescence ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; bcl-2-Associated X Protein ; genetics

The purpose was to study the responses of AML cell treated with cytochrome C and to explore the influence of cytochrome C on apoptosis of AML cell induced by daunorudicine (DNR). The differentiation of AML cell was detected by Wright-Giemsa staining and NBT test, the apoptosis of AML cell was assayed by flow cytometry and fluorescence microscopy. The results showed as follows: (1) different concentrations of cytochrome C could induce different effects on AML cells. Concentration of cytochrome C for differentiation was 10 microl/ml, for apoptosis was 20 microl/ml, and for necrosis was 40 microl/ml. (2) the apoptosis of AML cells decreased with the administration of cytochrome C in 10.0 microg/ml before treating AML cells with DNR (P < 0.01), but no change was shown with the administration of cytochrome C in 20.0 microg/ml (P > 0.05). (3) in reverse sequence, administrating of cytochrome C in 10 microl/ml and 20 microl/ml after treating AML cells with DNR, two different concentrations of cytochrome C could increase the apoptosis of AML cells (P < 0.01). It is suggested that cytochrome C may probably affect the apoptosis of AML cells induced by DNR.
Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Cytochromes c ; pharmacology ; Daunorubicin ; pharmacology ; Dose-Response Relationship, Drug ; Drug Synergism ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; pathology ; Microscopy, Fluorescence ; Tumor Cells, Cultured

To study the effect of interleukin-15 (IL-15) on the proliferation, differentiation and apoptosis of MDS CD34(+) cells, CD34(+) cells of high enrichment were separated by MACS system, and cultured in liquid media with different concentration of IL-15 in treated group and without IL-15 in the control group. Apoptosis of hematopoietic precursors was assayed by propidium iodine staining and cell by FCM, and the other MDS CD34(+) cells were stained by cytochemical staining after culture. The results showed that after culture with IL-15 the proliferation and differentiation of MDS CD34(+) cells were obviously promoted. It was found the every lineage of mature cells developed, the expressions of cell surface antigens CD71, CD33 and CD19 all increased in the MDS CD34(+) cell treated with IL-15. It is suggested that IL-15 stimulates the proliferation and differentiation of MDS CD34(+) cells, and partly shows anti-apoptosis effects which may be applicable to the therapy MDS.
Antigens, CD ; immunology ; Antigens, CD19 ; immunology ; Antigens, CD34 ; immunology ; Antigens, Differentiation, Myelomonocytic ; immunology ; Apoptosis ; drug effects ; Bone Marrow Cells ; drug effects ; immunology ; pathology ; Cell Cycle ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Flow Cytometry ; Humans ; Interleukin-15 ; pharmacology ; Microscopy, Fluorescence ; Myelodysplastic Syndromes ; blood ; immunology ; pathology ; Receptors, Transferrin ; immunology ; Sialic Acid Binding Ig-like Lectin 3

OBJECTIVETo study the effects of cadmium on hepatocellular DNA damage, expression of proto-oncogenes c-myc, c-fos, and c-jun as well as apoptosis in rats.

METHODSCadmium chloride at the doses of 5, 10, and 20 micromol/kg was given to rats by i.p. and there were 5 male SD rats in each group. Hepatocellular DNA damage was measured by single cell gel electrophoresis (or comet assay), while expression of proto-oncogenes c-myc, c-fos, and c-jun in rat hepatocytes were measured by Northern dot hybridization. C-Myc, c-Fos, and c-Jun were detected with immuno-histochemical method. Hepatocellular apoptosis was determined by TUNEL (TdT-mediated dUTP Nick End Labelling) and flow cytometry.

RESULTSAt the doses of 5, 10, and 20 micromol/kg, cadmium chloride induced DNA damage in rat hepatocytes and the rates of comet cells were 50.20%, 88.40%, and 93.80%, respectively. Results also showed an obvious dose-response relationship between the rates of comet cells and the dose of cadmium chloride (r = 0.9172, P < 0.01). Cadmium chloride at the doses of 5, 10, and 20 micromol/kg induced expression of proto-oncogenes c-myc, c-fos, and c-jun. The positive brown-yellow signal for c-myc, c-fos, and c-jun was mainly located in the cytoplasm of hepatocytes with immunohistochemical method. TUNEL-positive cells were detected in cadmium-treated rat livers. Apoptotic rates (%) of cadmium-treated liver cells at the doses of 5, 10, and 20 micromol/kg were (17.24 +/- 2.98), (20.58 +/- 1.35), and (24.06 +/- 1.77) respectively, being significantly higher than those in the control. The results also displayed an obvious dose-response relationship between apoptotic rates and the dose of cadmium chloride (r = 0.8619, P < 0.05).

CONCLUSIONCadmium at 5-20 micromol/kg can induce hepatocellular DNA damage, expression of proto-oncogenes c-myc, c-fos, and c-jun as well as apoptosis in rats.

Animals ; Apoptosis ; drug effects ; Cadmium ; toxicity ; DNA Damage ; Gene Expression Regulation ; drug effects ; Hepatocytes ; cytology ; drug effects ; metabolism ; Male ; Proto-Oncogene Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-fos ; genetics ; metabolism ; Proto-Oncogene Proteins c-jun ; genetics ; metabolism ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

The aim of this study was to investigate the effects of tyrosine kinase inhibitor PTK787 on cell proliferation, cell cycle and the expression of fak mRNA of human chronic myeloid leukemia (CML) cell line K562, and to explore the mechanism of PTK787 against acute myeloid leukemia. The MTT method was used to detect the effects of PTK787 in various concentrations and at different time points on proliferation of K562 cells; the flow cytometry was used to determine the effects of PTK787 in different concentrations on cell cycle of K562 cells; the RT-PCR was used to assay the expression of fak mRNA in K562 cells treated with PTK787 for 48 hours. The results showed that along with increasing of the concentration and prolonging of time, the inhibitory rate of PTK787 on K562 proliferation was gradually enhanced. The comparison between various concentration groups at same time or comparison between various time groups in same concentration showed significant differences (p < 0.05), in which the effect of 320 micromol/L PTK787 on cells was strongest, while the continuous increase of PTK787 concentration or prolong of action time did not enhance the inhibitory rate on K562 proliferation. With increasing of drug concentration, the cell proportion in G(1) phase gradually increased, the cell proportion in S phase gradually decreased, the comparison between various groups revealed significant differences (p < 0.05), however the continuous increase of drug concentration from 160 micromol/L did not obviously change the cell proportion in phases of cell cycle. With increasing of drug concentration, the expression of fak mRNA in K562 cells gradually reduced with significant differences between various groups (p < 0.05), but with continuous increase of drug concentration from 160 micromol/L, the effect of PTK787 on the expression of fak mRNA in K562 cells also did not obviously change. It is concluded that the PTK787 shows effect of anti-leukemia cells through inhibiting transformation of the K562 cells from G(1) phase into S phase and decreasing the expression of fak mRNA in cells.
Cell Cycle ; Cell Proliferation ; drug effects ; Focal Adhesion Kinase 1 ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Phthalazines ; pharmacology ; Pyridines ; pharmacology ; RNA, Messenger ; genetics

The objective of this study was to detect the level of plasma stromal cell-derived factor-1 (SDF-1) and the expression of CXCR4 (SDF-1 receptor in bone marrow cells) in children with Acute Leukemia (AL) and to investigate the relationship between the expression of CXCR4 and extramedullary infiltration. 48 children with acute leukemia and 20 with non-hematologic malignancies were selected into the AL group and the control group respectively. The peripheral plasma and bone marrow cells were collected. The level of SDF-1 in peripheral plasma was detected by ELISA and the expression of CXCR4 in bone marrow cells was determined by flow cytometry. The results showed that the levels of SDF-1 in peripheral plasma and the expression of CXCR4 in bone marrow cells of AL group was significantly higher than that of control group, among which the level of SDF-1 of the acute lymphoblastic leukemia (ALL) group was also higher than that of the acute myeloid leukemia (AML) group, the expression level of CXCR4 in the bone marrow cells of the extramedullary infiltration (EI) group was higher than that of the non-extramedullary Infiltration (NI) group, and all the differences between the both groups were significant. It is concluded that SDF-1 and CXCR4 express a high level in children with AL, which closely relates with the type of leukemia and the migration and infiltration of leukemia cells in bone marrow.
Acute Disease ; Adolescent ; Bone Marrow ; metabolism ; Bone Marrow Cells ; metabolism ; Case-Control Studies ; Chemokine CXCL12 ; blood ; metabolism ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Receptors, CXCR4 ; metabolism