ABSTRACT
OBJECTIVE@#To establish an animal model of acute B lymphoblastic leukemia (B-ALL) with minimal residual disease.@*METHODS@#The transplanted tumor was formed by subcutaneous injection of 2×107 Nalm-6 cells, and the body weight, activity status and tumor formation status of nude mice were observed. Peripheral blood, bone marrow, liver and spleen and other tissues of nude mice were taken for pathological examination to understand whether the success of subcutaneous modeling was accompanied by systemic metastasis.@*RESULTS@#There were 2×107 Nalm-6 cells injected subcutaneously in nude mice, (11.0±2.5) days later, the tumors of (3-4) × (3-4) mm were observed, the body weight of the nude mice was reduced and activity showed no limited. Infiltration of tumor cells in liver, spleen and bone marrow were observed in pathological sections.@*CONCLUSION@#The animal model of subcutaneous tumor of B-ALL was successfully established in nude mice.
Subject(s)
Animals , Humans , Mice , Body Weight , Disease Models, Animal , Mice, Nude , Neoplasm Transplantation , Neoplasm, Residual , Precursor Cell Lymphoblastic Leukemia-LymphomaABSTRACT
OBJECTIVE@#To study the effect of 280 nm-LED ultraviolet irradiation on the proliferation of acute promyelocytic leukemia (APL) HL-60 cells under hypoxic conditions and related mechanism.@*METHODS@#HL-60 cells in the logarithmic growth phase were selected and divided into control, hypoxia, ultraviolet and hypoxia+ultraviolet groups. The cells in the hypoxia group were treated with cobalt chloride (with a final concentration of 150 μmol/L), those in the ultraviolet group were irradiated by 280 nm-LED ultraviolet with an energy intensity of 30 J/m, and those in the hypoxia+ultraviolet group were treated with cobalt chloride and then irradiated by 280 nm-LED ultraviolet. After 48 hours of treatment, the cells were placed under an invert microscope to observe cell morphology. CCK-8 assay was used to measure the inhibition rate of cell proliferation. Annexin V-FITC/PI double staining flow cytometry was used to evaluate cell apoptosis. Quantitative real-time PCR was used to measure the mRNA expression of Bcl-2. Each experiment above was repeated three times independently.@*RESULTS@#Compared with the control group, the experimental groups showed shrinkage, decreased brightness, and disordered arrangement of cells, and the number of cells decreased over the time of culture. There were significant differences in the inhibition rate of cell proliferation and cell apoptosis rate among the groups (P<0.01), and the hypoxia+ultraviolet group showed the strongest inhibition of cell proliferation and induction of cell apoptosis, followed by the ultraviolet group and the hypoxia group. Compared with the control group, the other three groups had a gradual reduction in the mRNA expression of Bcl-2, and the hypoxia+ultraviolet group had a significantly greater reduction than the hypoxia and ultraviolet groups (P<0.01).@*CONCLUSIONS@#Both hypoxia and ultraviolet irradiation can inhibit the proliferation of HL-60 cells and induce cell apoptosis, and ultraviolet irradiation has a better effect on proliferation inhibition and cell apoptosis under hypoxic conditions than under normoxic conditions, possibly by downregulating the mRNA expression of Bcl-2.
Subject(s)
Humans , Apoptosis , Cell Hypoxia , Cell Proliferation , Leukemia, Promyelocytic, AcuteABSTRACT
OBJECTIVE@#To discuss the effect of insulin and metformin on a methylation and glycolipid metabolism of peroxisome proliferator-activated receptor γ coactivator-1A (PPARGC1A) of rat offspring with gestational diabetes mellitus (GDM).@*METHODS@#A total of 45 pregnant rats received the intraperitoneal injection of streptozotocin to establish the pregnant rat model of GDM. A total of 21 pregnant rats with GDM were randomly divided into three groups, with 7 rats in each group, namely the insulin group, metformin group and control group. Rats in the insulin group received the abdominal subcutaneous injection of 1 mL/kg recombinant insulin glargine at 18:00 every day. Rats in the metformin group received the intragastric infusion of metformin hydrochloride at 18:00 every day, with the first dose of 300 mg/kg. The doses of two groups were adjusted every 3 d to maintain the blood glucose level at 2.65-7.62 mmol/L. Rats in the control group received the intragastric infusion of 1 mL normal saline at 18:00 every day. After the natural delivery of pregnant rats, 10 offspring rats were randomly selected from each group. At birth, 4 wk and 8 wk after the birth of offspring rats, the weight of offspring rats was measured. The blood glucose level of offspring rats was measured at 4 wk and 8 wk, while the level of serum insulin, triglyceride and leptin was measured at 8 wk.@*RESULTS@#The weight of offspring rats at birth in the insulin group and metformin group was significantly lower than the one in the control group (P 0.05). The fasting blood glucose and random blood glucose in the insulin group and metformin group at 4 wk and 8 wk were all significantly lower than ones in the control group (P 0.05). The expression of PPARGC1A mRNA in the insulin group and metformin group was significantly higher and the methylation level of PPARGC1A was significantly lower than the one in the control group (P 0.05). Insulin and leptin at 8 wk in the insulin group and metformin group were significantly higher, while triglyceride was significantly lower than the one in the control group (P 0.05).@*CONCLUSIONS@#GDM can induce the methylation of PPARGC1A of offspring rats to reduce the expression of PPARGC1A mRNA and then cause the disorder of glycolipid metabolism when the offspring rats grow up; the insulin or metformin in the treatment of pregnant rats with GDM can reduce the methylation level of PPARGC1A and thus improve the abnormal glycolipid metabolism of offspring rats.
ABSTRACT
Objective: To discuss the effect of insulin and metformin on a methylation and glycolipid metabolism of peroxisome proliferator-activated receptor γ coactivator-1A (PPARGC1A) of rat offspring with gestational diabetes mellitus (GDM). Methods: A total of 45 pregnant rats received the intraperitoneal injection of streptozotocin to establish the pregnant rat model of GDM. A total of 21 pregnant rats with GDM were randomly divided into three groups, with 7 rats in each group, namely the insulin group, metformin group and control group. Rats in the insulin group received the abdominal subcutaneous injection of 1 mL/kg recombinant insulin glargine at 18:00 every day. Rats in the metformin group received the intragastric infusion of metformin hydrochloride at 18:00 every day, with the first dose of 300 mg/kg. The doses of two groups were adjusted every 3 d to maintain the blood glucose level at 2.65-7.62 mmol/L. Rats in the control group received the intragastric infusion of 1 mL normal saline at 18:00 every day. After the natural delivery of pregnant rats, 10 offspring rats were randomly selected from each group. At birth, 4 wk and 8 wk after the birth of offspring rats, the weight of offspring rats was measured. The blood glucose level of offspring rats was measured at 4 wk and 8 wk, while the level of serum insulin, triglyceride and leptin was measured at 8 wk. Results: The weight of offspring rats at birth in the insulin group and metformin group was significantly lower than the one in the control group (P 0.05). The fasting blood glucose and random blood glucose in the insulin group and metformin group at 4 wk and 8 wk were all significantly lower than ones in the control group (P 0.05). The expression of PPARGC1A mRNA in the insulin group and metformin group was significantly higher and the methylation level of PPARGC1A was significantly lower than the one in the control group (P 0.05). Insulin and leptin at 8 wk in the insulin group and metformin group were significantly higher, while triglyceride was significantly lower than the one in the control group (P 0.05). Conclusions: GDM can induce the methylation of PPARGC1A of offspring rats to reduce the expression of PPARGC1A mRNA and then cause the disorder of glycolipid metabolism when the offspring rats grow up; the insulin or metformin in the treatment of pregnant rats with GDM can reduce the methylation level of PPARGC1A and thus improve the abnormal glycolipid metabolism of offspring rats.
ABSTRACT
<p><b>OBJECTIVE</b>To retrospectively analyze the clinical characteristics and the treatment outcomes of older children with acute lymphoblastic leukemia (ALL), and to evaluate the multicenter cooperation regimen (ALL-2005).</p><p><b>METHODS</b>The clinical data of 103 newly diagnosed ALL children aged 10 to 18 years old from five hospitals were enrolled in this study. They were all received ALL-2005 protocol. The clinical characteristics, the event-free survival (EFS), the overall survival (OS) and the prognostic analysis were evaluated.</p><p><b>RESULTS</b>(1) Of the 103 patients, 62 were boys and 41 girls, with a median age of 12.3 years old. According to immunophenotyping, 90 (87.4% ) of 103 patients were diagnosed as B-ALL and 13 (12.6%) as T-ALL. According to risk factor, 65 (63.1%) were in intermediate risk group (MR-ALL) and 38 (36.9%) in high risk group (HR-ALL). Central nervous system leukemia (CNSL) happened in 4 (3.9%) patients at diagnosis. Of the 89 patients received chromosome test, 58 (65.2%) obtained the test results, including 21(36.2%) with aberrational chromosomes and 37 (63.8%) with normal karyotype. Of 81 patients received molecular biological test, 16 (19.8%) were positive for fusion gene. (2) After induction therapy, 97 (94.2%) obtained complete remission (CR). Twenty-eight patients relapsed with a median time of 11.9 months (ranged 2.9-57.8 months), and 38 (36.9%) patients died during the treatment. As of September 30, 2012, the median follow-up was 47 months (ranged 0.4-92.6 months). The 5-year EFS and 5-year OS of ALL patients were (60.2 ± 4.8)% and(64.1 ± 4.7)%. The 5-year EFS of MR-ALL and HR-ALL were (73.8 ± 5.5)% and (31.6 ± 8.3)% (P<0.01), the 5-year OS of MR- ALL and HR-ALL were (78.5 ± 5.1)% and (35.9 ± 8.0)% (P<0.01), respectively. (3) Cox proportion hazard regression model analysis indicated that age of 14-18 years old and BCR- ABL translocation or t(9;22) were independent risk prognostic factor for 5-year EFS.</p><p><b>CONCLUSION</b>The incidence and prognosis in older childhood ALL were related with age, risk and biological characteristics. BCR-ABL translocation or t(9;22) was the risk factor of prognosis. ALL- 2005 protocol was recommended as the regimen for older childhood ALL.</p>
Subject(s)
Adolescent , Child , Female , Humans , Male , Disease-Free Survival , Incidence , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Diagnosis , Therapeutics , Prognosis , Risk Factors , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To study the effect of bacillus Callmette-Guérin (BCG) on cytotxicity of cytotoxic T lymphocyte (CTL) from human peripheral blood of children with acute lymphoblastic leukemia (ALL) for killing HL-60 cells in vitro.</p><p><b>METHODS</b>The mononuclear cells were isolated from peripheral blood of ALL children and healthy children, and were cultured with RPMI1640, interleukin-2 (IL-2), phytohemagglutinin (PHA) and BCG.The growth of CTLs was observed by light microscopy. The proportions of CD3, CD3+CD4+ and CD3+CD8+ were determined by flow cytometry 10 days after culture. MTT method was performed to detect the cytotoxicity of CTLs for killing HL-60 cells.</p><p><b>RESULTS</b>Neither the cell number nor the volume of CTLs changed significantly within 2 days of culture, but both began increasing on the 3rd day of culture and reached a peak on the 6-10th days. On the 10th day of culture, the cell number of CTLs in the BCG treatment group was much higher than in the group without BCG treatment. The CD3+CD8+ proportion in the leukemia group was much higher than in the control group. With the effect of BCG, the CD3+CD8+ proportion of the two groups became much higher. The cytotoxicity of CTLs for killing HL-60 cells in the leukemia group was weaker than in the control group.</p><p><b>CONCLUSIONS</b>BCG along with IL-2 and PHA promotes the proliferation of CTLs and enhances the ability of CTLs in killing HL-60 cells.</p>
Subject(s)
Child, Preschool , Female , Humans , Male , Cytotoxicity, Immunologic , HL-60 Cells , Interleukin-2 , Pharmacology , Lymphocyte Activation , Mycobacterium bovis , Allergy and Immunology , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Allergy and Immunology , T-Lymphocytes, Cytotoxic , Allergy and ImmunologyABSTRACT
This study was purposed to investigate the effect of crocin on the proliferation in vitro and immune function of dendritic cells (DC) derived from the bone marrow of children with acute leukemia. The mononuclear cells were isolated from bone marrow of leukemia children by Ficoll-Hypaque. The experiment was divided into six groups: blank control group (A), crocin 1.25 mg/ml group (B), cytokines (rhGM-CSF 75 ng/ml+rhIL-4 75 ng/ml+rhTNF-α 50 ng/ml) group (C), cytokines+crocin 0.3125, 1.25 or 5.0 mg/ml groups (D, E, F). The numbers of DC were counted and the phenotypes of DC were determined by flow cytometry on the ninth day of culture. The DC of different groups were mixed with T cells just separated from peripheral blood of another children with acute lymphoblastic leukemia, and cultured with rhIL-2 200 U/ml for 5 d. The function of DC was detected by mixed lymphocyte reaction (MLR). The results indicated that the test groups and control group all obtained a certain amount of typical DC, but the DC numbers in test groups were all higher than those in control group (P < 0.01). Cultured for 9 days, the rates of CD1a(+), CD83(+), and HLA-DR(+) in group C, D, E, F were higher than group A (P < 0.01). There was no statistically significant difference between A and B groups (P > 0.05). MLR showed that with the increasing of DC, the stimulation index of T cells in group A and B was not rising (P > 0.05); the stimulated index of T cells in group C and E was significantly rising, there was statistically significant difference between them (P < 0.01). When the number of stimulated cells was the same, the stimulation index of T cell in group E was the highest (P < 0.01). It is concluded that the capability of DC proliferation promoted by crocin alone is lower than that of its combination with rhGM-CSF, rhIL-4 and rhTNF-α, but the crocin can synergically promote the maturity of DC cooperating with rhGM-CSF, rhIL-4 and rhTNF-α. The DC induced by crocin can particularly enhance the proliferation of T cells.
Subject(s)
Child , Humans , Bone Marrow Cells , Cell Biology , Carotenoids , Pharmacology , Cell Proliferation , Dendritic Cells , Cell Biology , Leukemia , Pathology , Lymphocyte Culture Test, Mixed , T-Lymphocytes , Cell Biology , Tumor Cells, CulturedABSTRACT
This study was purposed to explore the anti-leukemia effect of bacillus calmette-guerin vaccine (BCG) on the human myeloid leukemia cell xenograft models. An animal model was established by inoculating the human myeloid leukemia HL-60 cells into the BALB/c (8 - 10 weeks of age) nude mice. The mice were randomly divided into two groups: control group and experimental group. Nude mice in control group were injected with physiological saline, while those of experimental group were given BCG and inactivated BCG respectively. The tumor growth was assayed by using caliper. The survival time of nude mice was determined. Necrotic extent and morphological changes of tumor were observed and examined by HE staining and immunohistochemical method. The results indicated that on 5th-7th days after tumor inoculated, 2 - 3 mm tumor mass could be observed. The tumor volume increased over the time. HE staining of tumor tissues showed that there were different degrees of tumor necrosis in BCG group and inactivated BCG group. Immunohistochemistry results demonstrated that CD20 positive cells were obviously observed in the necrotic area of BCG group, compared with the control group and inactivated BCG group. It is concluded that human myeloid leukemia HL-60 cells have been successfully transplanted in nude mice, and the systemic metastasis occurs along with the prolongation of time. BCG inoculation can delay the tumor growth and prolong the survival time of nude mice with leukemia, suggesting that BCG has an antitumor effect.
Subject(s)
Animals , Humans , Mice , BCG Vaccine , Therapeutic Uses , HL-60 Cells , Leukemia, Myeloid , Therapeutics , Mice, Inbred BALB C , Mice, Nude , Xenograft Model Antitumor AssaysABSTRACT
This study was to investigate the proliferative inhibition and apoptosis of human leukemia HL-60 cells induced by crocin and their possible mechanisms. The cell viability was tested by cell counting. The morphology of HL-60 cells was observed by fluorescence microscopy. The MTT assay was used to evaluate the inhibitory effect of crocin on the growth of HL-60 cells. Flow cytometry was used to measure the cell cycle. RT-PCR was used to detect bcl-2 and bax expression. The results indicated that the growth of HL-60 cells was inhibited remarkably in the dose and time dependent way. When the crocin concentration was higher than 5 mg/ml, the percentage of apoptotic HL-60 cells was not increased, on the contrary this percentage decreased, the cells manifested necrosis. Flow cytometry profiles revealed that cells were blocked in G₀/G₁ phase, the cell proliferation was inhibited obviously at 5 mg/ml. RT-PCR detection revealed that the expression of bcl-2 was down-regulated strikingly and bax was up-regulated. It is concluded that the crocin can inhibit the proliferation of HL-60 cells effectively, and block cells in G₀/G₁ phase. The mechanisms by which crocin induced apoptosis in HL-60 cells may be related to the inhibition of bcl-2 and activation of bax.
Subject(s)
Humans , Apoptosis , Carotenoids , Pharmacology , Therapeutic Uses , Cell Cycle , Cell Proliferation , Gene Expression Regulation, Leukemic , HL-60 Cells , Leukemia , Drug Therapy , Metabolism , Phytotherapy , Proto-Oncogene Proteins c-bcl-2 , Metabolism , bcl-2-Associated X Protein , MetabolismABSTRACT
The aim of this study was to explore the effect of muramyl dipeptide (MDP) on proliferation of dendritic cells (DCs) from bone marrow of children with acute leukemia in vitro. The mononuclear cells were isolated from bone marrow of children with acute leukemia to induce dendritic cells. The experiment was divided into 4 groups. The control group: MNC + RPMI 1640 medium; test group 1: MNC + MDP; test group 2: MNC + rhGM-CSF + IL-4 + rhTNFα; test group 3: MNC + rhGM-CSF + IL-4 + rhTNFα + MDP. The growth of DCs was observed by inverted microscope every day; the number of DCs in different groups were counted, the immunophenotypes of DCs were detected by flow cytometry on day 8 of culture. The results indicated that a certain number of typical DCs could be detected in all experimental groups. The DC number in control and 3 test groups were (0.85 ± 0.23) x 10⁵/L, (2.31 ± 0.24) x 10⁵/L, (3.26 ± 0.37) x 10⁵/L and (4.16 ± 0.34) x 10⁵/L, respectively, among which DC number is in all 3 test groups were higher than that in control group (p < 0.01), the DC number in test group 1 was lower than that in test groups 2 and 3 (p < 0.01), while it in test group 3 was higher than that in test group 2 (p < 0.01). The percentages of HLA-DR in control, test group 1, 2 and 3 were 19.98 ± 3.74, 37.24 ± 4.32, 58.81 ± 2.08 and 77.48 ± 5.57 respectively; the percentages of CD1a and CD83 in control, test group 1, 2 and 3 were 11.46 ± 2.43, 28.71 ± 6.64, 46.92 ± 4.78 and 57.03 ± 3.07, as well as 13.05 ± 5.70, 36.32 ± 5.61, 54.95 ± 7.83 and 75.70 ± 6.67 respectively. The comparison of HLA-DR, CD1a and CD83 levels in control and test group 1, 2 showed that their results were consistent with results of DC numbers. It is concluded that MDP not only promotes the proliferation of DCs derived from bone marrow of children with acute leukemia in vitro, cooperates with rhGM-CSF, rhIL-4 and rhTNFalpha in promoting of the proliferation and maturation of DCs, while the promotive effect of MDP alone on the proliferation of DCs is not as good as its combination with cytokines.
Subject(s)
Child , Humans , Acetylmuramyl-Alanyl-Isoglutamine , Pharmacology , Bone Marrow Cells , Cell Biology , Cell Proliferation , Cells, Cultured , Dendritic Cells , Cell Biology , Flow Cytometry , Leukemia , Allergy and Immunology , Pathology , Tumor Cells, CulturedABSTRACT
This study was purposed to investigate the effect of bacillus calmette-guerin (BCG) on the expansion of human dendritic cells (DC) from peripheral blood of pediatric patients with leukemia in vitro. The experiment was divi-ded into two groups: the control and the test group, and the latter group was divided into 3 subgroups: BCG (only BCG), GTI (GM-CSF, TNF-α, IL-4) and GTIB (GM-CSF, TNF-α, IL-4 plus BCG). On day 9 of culture the DCs were counted in each groups, the phenotypes of DC were determined by flow cytometry and these DC were stained with Wright-Giemsa for observation and photography under microscopy. The results showed that the test groups all obtained a certain amount of typical DC; the number of DC in the BCG subgroup is lower than that in the GTI and GTIB subgroups (t=4.20; 6.36, p<0.01); there was no significant difference between the GTI and the GTIB subgroups (t=2.25; p>0.05). The rate of CD1a+ in the BCG subgroup was obviously higher than that in the control group (t=3.04, p<0.05), but was lower than that in the GTI and the GTIB subgroups (t=2.79, 6.41, p<0.05), there was no significant difference between the GTI and the GTIB subgroups (t=0.65, p>0.05). The rate of HLA-DR+, CD83+ in the BCG group was higher than that in the control group (t=4.77, 4.15; p<0.05), but lower than that in the GTI and the GTIB subgroups (t=6.65, 3.19; p<0.05). The rate of HLA-DR+, CD83+ in the GTI subgroup was lower than that in the GTIB subgroup (t=5.64, 2.98; p<0.05). It is concluded that BCG not only promotes the proliferation of DC derived from human peripheral blood of leukemia patients in vitro, but also cooperates with rhGM-CSF, rhTNF-α and rhIL-4 in promoting the maturation of DCs.
Subject(s)
Child , Humans , BCG Vaccine , Allergy and Immunology , Pharmacology , Cell Differentiation , Cells, Cultured , Dendritic Cells , Cell Biology , Leukemia , Allergy and Immunology , Mycobacterium bovis , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To study the effects of strychnos alkaloids on the proliferation of adult rat neuroprogenitor cells.</p><p><b>METHODS</b>Strychnos alkaloids free of strychnine and brucine were extracted from Strychnos nux vomica, and the effects of Strychnos alkaloids on the survival of HEK293 and PC12 cells were evaluated using MTT assay. In vitro cultured adult rat neuroprogenitor cells isolated from the hippocampus were treated with different concentrations of Strychnos alkaloids for 2 days, and the cell proliferation was assessed using BrdU incorporation assay.</p><p><b>RESULTS</b>At the concentration above 0.5 mg/ml, Strychnos alkaloids produced toxic effect against HEK293 cells (P<0.0001), while for PC12 cells, Strychnos alkaloids inhibited the cell survival at the concentration as low as 5 microg/ml (P<0.0001). After 2 days of exposure to 50 microg/ml Strychnos alkaloids, the neuroprogenitor cells showed significantly decreased number of BrdU-positive cells (P<0.01), but the total cell number remained stable when compared with that of the control cells (P>0.05), whereas at the concentration of 100 microg/ml, Strychnos alkaloids produced obvious cytotoxicity against the neuroprogenitor cells.</p><p><b>CONCLUSION</b>Strychnos alkaloids can significantly inhibit the proliferation of adult rat neuroprogenitor cells, and this effect is probably selective, suggesting the potential of Strychnos alkaloids as a new drug for treatment of neurocytoma.</p>
Subject(s)
Animals , Humans , Rats , Alkaloids , Pharmacology , Cell Line , Cell Proliferation , Drugs, Chinese Herbal , Pharmacology , Hippocampus , Cell Biology , Neurons , Cell Biology , PC12 Cells , Cell Biology , Stem Cells , Cell Biology , Strychnos , ChemistryABSTRACT
Paeoniflorin is one of the bioactive components of Paeonia lactiflora, a traditional Chinese herbal medicine. It is the main monoterpene glucoside isolated from the P. lactiflora in 1963. Since then, researchers have found that paeoniflorin has multifold pharmacological effects. In this review, based on the recent available papers published in PubMed and National Knowledge Infrastructure Data Base, we present the major current approaches in understanding the detection methodology, pharmacokinetics and pharmacology, and toxicology of paeoniflorin.
Subject(s)
Animals , Humans , Benzoates , Pharmacokinetics , Pharmacology , Bridged-Ring Compounds , Pharmacokinetics , Pharmacology , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Pharmacokinetics , Pharmacology , Enzyme-Linked Immunosorbent Assay , Glucosides , Pharmacokinetics , Pharmacology , MonoterpenesABSTRACT
To investigate the effects of interferon alpha-2b on proliferation and apoptosis in HL-60 cells, HL-60 cells were cultured in different concentrations of IFN alpha-2b. The morphologic changes were observed by Wright's and acridine orange (AO) and ethidium bromide (EB) staining respectively. Inhibition of proliferation was detected by MTT. Expression of CD13(+) was checked by indirect fluoroimmunoassay. The results showed that apoptosis rate of HL-60 cells assayed by the above-mentioned two methods was (51 +/- 2)% and (78 +/- 3)% respectively and OD(570) values of proliferation inhibited were 1.8 +/- 0.1 and 1.0 +/- 0.1 respectively when the concentrations of the IFN(alpha-2b) were 500 and 10,000 U/ml in culture for 48 hours. Morphology and count of CD13(+) cells were changed. CD13(+) cell expression rate was (62 +/- 2)% and (30 +/- 3)% respectively when the concentrations of the IFN(alpha-2b) were 500 and 10,000 U/ml in culture for 48 hours. It is concluded that IFN(alpha-2b) can enhance the apoptosis of HL-60 cells, inhibit their proliferation, promote their maturation and differentiation.
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Apoptosis , CD13 Antigens , Genetics , Cell Proliferation , HL-60 Cells , Interferon-alpha , Pharmacology , Recombinant ProteinsABSTRACT
The aim was to study the mechanisms of HL-60 cell apoptosis induced by nimodipine (NMDP) and cytarabine (Ara-C). The DNA fragment was detected by agarose gel electrophoresis. The expressions of bcl-2 and bax gene proteins related with apoptosis were investigated by immunohistochemistry. The results showed that HL-60 cell apoptosis rate had been increasing in the experimental groups compared with the control group since culturing 8 hours. The expression of Bcl-2 protein was lower and the expression of Bax protein was higher in the experimental groups than that in the control group, while ratio of bcl-2/bax was lower in the experimental groups than that in the control group. It is concluded that NMDP and Ara-C induce apoptosis of HL-60 cells, and the mechanism of apoptosis induced by them may down-regulate the expression of bcl-2 gene and up-regulate the expression of bax gene. The mechanism of HL-60 cell apoptosis induced by Ara-C and NMDP is probably associated with the down-regulation of Bcl-2 protein expression.
Subject(s)
Humans , Antimetabolites, Antineoplastic , Pharmacology , Apoptosis , Cytarabine , Pharmacology , Drug Resistance, Neoplasm , Drug Synergism , Gene Expression Regulation, Neoplastic , HL-60 Cells , Nimodipine , Pharmacology , Proto-Oncogene Proteins c-bcl-2 , Genetics , bcl-X Protein , GeneticsABSTRACT
This study was aimed to investigate the changes of mitochondrial membrane potential (DeltaPsim) of HL-60 cells induced by cytarabine and the correlation between mitochondrial membrane potential and apoptosis of HL-60 cells. HL-60 cells were stained with Rhodamine 123; change of mitochondrial membrane potential of HL-60 cells was detected by flow cytometry. AO/EB staining and flow cytometry were used to examine the apoptosis of HL-60 cells. The results showed that the levels of HL-60 cell DeltaPsim in experimental groups decreased after cultured for 6 hours. In Ara-C 0.05 mg/ml group, rhodamine 123 fluorescence intensity in mitochondria of HL-60 cells at 6, 12, 24 hours were 117.9+/-7.6, 100.9+/-7.7, 87.6+/-10.7, respectively, there was significant difference between the different culture groups (p<0.05). In Ara-C 0.1 mg/ml group, rhodamine 123 fluorescence intensity in mitochondria of HL-60 cells at 6, 12, 24 hours were 111.9+/-10.1, 86.6+/-9.2, 68.4+/-12.2, respectively, there was significant difference between the different culture groups (p<0.05); rhodamine 123 fluorescence intensity was significantly different between the two groups at 12, 24 hours (p<0.05). In Ara-C 0.05 mg/ml group, the apoptosis rate of HL-60 cells at 6, 12, 24 hours were (41.2+/-3.0)%, (53.7+/-5.1)%, (65.8+/-2.6)% respectively, there was significant difference between the different culture groups (p<0.01); In Ara-C 0.1 mg/ml group, the apoptosis rate of HL-60 cells at 6, 12, 24 hours were (45.7+/-4.1)%, (58.2+/-4.3)%, (70.1+/-2.3)% respectively, there was significant difference between the different culture groups (p<0.01); the apoptosis rates showed no significantly difference between the two groups at same time. The changes of mitochondrial membrane potential and apoptosis rate of HL-60 cells were significantly negatively correlated. In Ara-C 0.05 mg/ml group, r was -0.89, p<0.01, while in Ara-C 0.1 mg/ml group, r was -0.76, p<0.01. It is concluded that the mitochondrial membrane potential on HL-60 cells decrease at HL-60 cells apoptosis induced by Ara-C, therefore the reduction of mitochondrial membrane potential may be one of the important mechanisms at the induced apoptosis.
Subject(s)
Humans , Antimetabolites, Antineoplastic , Pharmacology , Apoptosis , Cytarabine , Pharmacology , HL-60 Cells , Membrane Potential, MitochondrialABSTRACT
To investigate the effect of mycobacterium phlei F.U.36 suspended liquor (Utilin"s", U) on the culture and proliferation of dendritic cells (DCs) derived from human umbilical cord blood in vitro, the mononuclear cells (MNCs) were isolated from human umbilical cord blood and cultured with RPMI 1640 in the control group. Test groups consisted of Utilin"s" group (only Utilin"s"), GTI group (GM-CSF, TNF-alpha, IL-4) and GTIU group (GM-CSF, TNF-alpha, IL-4 and Utilin"s"). MNCs in all test groups were cultured with RPMI-1640. The growth of DCs was observed by the light microscopy, the phenotypes of DCs were determined by flow cytometry on the 10th day of culture, and some harvest cells were stained with Wright-Giemsa, then observed and photographed under the oil immersion objective. The results showed that the test groups all displayed some number of typical DCs; both CD1a positive cell rate and HLA-DR positive cell rate of the Utilin"s" group were higher than those of the control; HLA-DR positive cell rate of GTIU group increased most significantly and much higher than that of the GTI group. It is concluded that mycobacterium phlei F.U.36 not only promotes the proliferation of DCs derived from human umbilical cord blood in vitro, but also co-operates with rhGM-CSF, rhTNF-alpha and rhIL-4 in promoting the maturity of DCs.
Subject(s)
Humans , Cell Proliferation , Cells, Cultured , Dendritic Cells , Cell Biology , Fetal Blood , Cell Biology , Mycobacterium phlei , PhysiologyABSTRACT
Dendritic cells (DCs) derived from bone marrow cells are specialized cells for the uptake, processing, and presentation of foreign and self-antigens. The study indicated that re-transfusion of DCs pulsed with tumor-associated antigen can induce an vigorous specific anti-tumor response in clinic. The present study was aimed to investigate the enhancing effect of DCs derived from human cord blood on T cells in killing tumor cells. Human cord blood mononuclear cells were isolated from human cord blood by density gradient centrifugation using lymphocyte separating medium, and cord blood mononuclear cells were obtained by adherence and cultured in a liquid culture system with GM-CSF and IL-4 for 15 days. Then the cells were analyzed for phenotypes of CD1a by indirect immunofluorescence. The capacity of DCs to initiate T cell-dependent anti-tumor immune responses was assayed by MTT kit. The ratios of DCs to tumor cells in experimental groups were 20:1, 50:1 and 100:1 respectively. The DCs were not added in control group. The results indicated that in the presence of GM-CSF and IL-4, the DCs with typical morphological features at days 15 were observed. At that time, (43.12 +/- 5.83)% CD1a(+) cells were obtained. In addition to these phenotypic properties, the DC of experimental groups could remarkably initiate T cell-dependent anti-tumor immune responses with different ratios compared with control group (P < 0.01), there were no significant difference of killing effects between 100:1 and 50:1 groups (P > 0.05), and killing effect of DC in 20:1 group was higher than that in 100:1 or 50:1 groups (P < 0.05). It is concluded that human cord blood mononuclear cells can serve as a better source of DC, which can promote the capacity to initiate T cell-dependent anti-tumor immune responses.
Subject(s)
Humans , Apoptosis , Allergy and Immunology , Brain Neoplasms , Pathology , Cell Differentiation , Cells, Cultured , Dendritic Cells , Cell Biology , Allergy and Immunology , Fetal Blood , Cell Biology , Granulocyte-Macrophage Colony-Stimulating Factor , Pharmacology , Interleukin-4 , Pharmacology , Leukocytes, Mononuclear , Cell Biology , Neuroblastoma , Pathology , Recombinant Proteins , T-Lymphocytes , Allergy and Immunology , Tumor Cells, CulturedABSTRACT
0.05),and NK cells function(CD56),humoral immunologic function(IgG,IgM,IgA)were significantly elevated after TP-5 administration for 3 months(Pa
ABSTRACT
Objective To explore influence of long-term inhaled glucocoticoids(IGS) on soluble intercellular adhesion molecule-1(sICAM-1)) in children with bronchial asthma.Methods Enzyme-linked immunosorbent assay(ELISA) method was used to detect the serum sICAM-1 level in 36 healthy children and 29 children with bronchial asthma(untreated and post-treated for 3,6 and 12 months).Results 1.Serum sICAM-1 level was significantly elevated in children with asthma and significantly higher than that in normal control group(P