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1.
Journal of Experimental Hematology ; (6): 77-85, 2021.
Article in Chinese | WPRIM | ID: wpr-880035

ABSTRACT

METHODS@#From January 2005 to December 2013, 83 patients with refractory/recurrent CD20@*RESULTS@#All the patient achieved complete response. The median follow.up time was 39 months. Both the two groups collected peripheral blood stem cells successfully, and had no difference in hematopoietic reconstitution time. Three patients in treatment group and six patients in control group relapsed and the three year overall survival and EFS in treatment group was significantly higher than that in control group, that is(93.0% vs 73.1%, P=0.037) and (89.5% vs 65.4%, P=0.034), respectively. Subgroup analysis showed that: compared with the treatment group in which using R in the whole courses(before and after transplantation, and collection of stem cells) was superior to the control group in both OS and EFS, with the OS 97% vs 87.5% (P>0.05) and EFS 97% vs 76.2% (P=0.05) respectively. While stratified by the different courses of rituximab, the OS was 88.9% (1-2 courses, 9 cases), 93.1% (3-4 courses, 29 cases), 94.7%(more than 5 courses,19 cases), and EFS was 77.8%, 89.7% and 94.7%, respectively.@*CONCLUSION@#For the patients with refractory/recurrent CD20


Subject(s)
Humans , Antineoplastic Combined Chemotherapy Protocols , Disease-Free Survival , Hematopoietic Stem Cell Transplantation , Hodgkin Disease , Lymphoma, Non-Hodgkin/drug therapy , Rituximab/therapeutic use , Transplantation, Autologous , Treatment Outcome
2.
Braz. j. med. biol. res ; 45(10): 906-912, Oct. 2012. ilus, tab
Article in English | LILACS | ID: lil-647750

ABSTRACT

Hypoxia inducible factor-1α (HIF-1α) is an important transcription factor, which plays a critical role in the formation of solid tumor and its microenviroment. The objective of the present study was to evaluate the expression and function of HIF-1α in human leukemia bone marrow stromal cells (BMSCs) and to identify the downstream targets of HIF-1α. HIF-1α expression was detected at both the RNA and protein levels using real-time PCR and immunohistochemistry, respectively. Vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1α (SDF-1α) were detected in stromal cells by enzyme-linked immunosorbent assay. HIF-1α was blocked by constructing the lentiviral RNAi vector system and infecting the BMSCs. The Jurkat cell/BMSC co-cultured system was constructed by putting the two cells into the same suitable cultured media and conditions. Cell adhesion and secretion functions of stromal cells were evaluated after transfection with the lentiviral RNAi vector of HIF-1α. Increased HIF-1α mRNA and protein was detected in the nucleus of the acute myeloblastic and acute lymphoblastic leukemia compared with normal BMSCs. The lentiviral RANi vector for HIF-1α was successfully constructed and was applied to block the expression of HIF-1α. When HIF-1α of BMSCs was blocked, the expression of VEGF and SDF-1 secreted by stromal cells were decreased. When HIF-1α was blocked, the co-cultured Jurkat cell’s adhesion and migration functions were also decreased. Taken together, these results suggest that HIF-1α acts as an important transcription factor and can significantly affect the secretion and adhesion functions of leukemia BMSCs.


Subject(s)
Humans , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Leukemia, T-Cell/metabolism , Mesenchymal Stem Cells/metabolism , Cell Adhesion , Enzyme-Linked Immunosorbent Assay , Immunohistochemistry , Jurkat Cells , Real-Time Polymerase Chain Reaction
3.
Chinese Journal of Hematology ; (12): 672-674, 2009.
Article in Chinese | WPRIM | ID: wpr-283919

ABSTRACT

<p><b>OBJECTIVE</b>To sum up the clinical experience of the diagnosis and treatment of intracerebral infiltration by monoclonal plasmacytoid cells in Waldenstrom's macroglobulinemia(Bing-Neel syndrome).</p><p><b>METHODS</b>The clinical data of the diagnosis and treatment of a case of Bing-Neel syndrome was analyzed.</p><p><b>RESULTS</b>A 56-year-old male was diagnosed as Waldenstrom's macroglobulinemia one year ago, and presented with persistent headache during the treatment period. Magnetic resonance imaging showed a high intensity area on T2-weighed images in the right frontal lobe which was well enhanced by gadolinium-diethylenetriaminepenta-acetic acid. Infiltration of neoplastic cells was confirmed by biopsy. Immunohistochemical examination showed that mature plasmacytoid cells in the cerebral parenchyma were immunoglobulin M positive.</p><p><b>CONCLUSION</b>Infiltration in CNS (Bing-Neel syndrome) is uncommon in Waldenstrom's macroglobulinemia. As there is no effective therapy for this Bing-Neel syndrome, combination of radiation and chemotherapy should be considered for this situation.</p>


Subject(s)
Humans , Male , Middle Aged , Brain , Pathology , Neoplasm Invasiveness , Waldenstrom Macroglobulinemia , Pathology
4.
Journal of Experimental Hematology ; (6): 1437-1441, 2008.
Article in Chinese | WPRIM | ID: wpr-234217

ABSTRACT

In order to study the influence of different gelatin concentrations, and lymphocyte isolation liquid on primary culture of umbilical cord blood-derived adhesive cells (hCBACs), the red blood cells of umbilical cord blood was separated by 3% and 6 % gelatin for detecting the effectiveness of sedimentation, then the adhesion rate at 48 hours, the day of initial expansion and the rate of culture success were detected for hCBACs cultured with CD34(+) cells after the mononuclear cells were separated by 6% gelatin followed by Ficoll and Percoll, and the morphological characteristics and growth status were observed by invert microscopy. Cytochemistry stain for nonspecific esterase stain (NSE), peroxidase (POX), periodic acid Schiff reaction (PAS) and alkali phosphatase (ALP) and immunocytochemistry labeling for CD31, CD45, CD68 and fibronectin (Fn) were detected. The results showed that 6 % gelatin was better than that 3% gelatin for red blood sedimentation. The Percoll was predominant over Ficoll in adhesion rate at 48 hours, the day of initial expansion, the time of initial formation of adhesive cell colony units, the time of maximal numbers of adhesive cell colony units, the the cell fusion time and ratio of culture success. 60% fibroblast-liked cells, 36% macrophage liked cells and 4% small-round cells were observed in cells isolated by both isolated methods. The cytochemistry stain for NSE, POX, PAS and ALP was similar in two groups, the difference was not statistically significant between these two groups. The immunocytochemistry labeling for CD31, CD45, CD68 and Fn was also similar in both groups and the difference was also not statistically significant between these two groups. It is concluded that the combination of 6% gelatin with Percoll is an ideal separation method for primary culture of hCBACs, which provides basic information for clinical application.


Subject(s)
Humans , Cell Separation , Methods , Cells, Cultured , Fetal Blood , Cell Biology , Gelatin , Pharmacology , Lymphocytes , Cell Biology
5.
Journal of Experimental Hematology ; (6): 147-150, 2008.
Article in Chinese | WPRIM | ID: wpr-253363

ABSTRACT

This study was aimed to investigate the expression of SDF-1 mRNA in SDF-1 cDNA-modified human bone marrow mesenchymal stem cells (hBMSCs) before and after transfection. The hBMSCs were isolated, cultured and identified, the SDF-1-pIRES2-EGFP eukaryotic expressing vector was constructed, and then the hBMSCs were transfected with the vector encapsulated by lipofectamine 2000. The transfection efficiency was measured by observing the expression of green fluorescence protein and detecting the mRNA by RT-PCR. The results indicated that the expression of SDF-1 mRNA increased by about 20% after hBMSCs were transfected instantaneously by SDF-1-pIRES2-EGFP. It is concluded that SDF-1 cDNA eukaryotic expression vector can be instantly transfected into hBMSCs by lipofectamine 2000, but the efficiency was too low to obtain enough steady transferred hBMSCs. Other procedures should be trialed to improve the transfection efficiency.


Subject(s)
Humans , Bone Marrow Cells , Cell Biology , Chemokine CXCL12 , Genetics , Metabolism , DNA, Complementary , Genetics , Genetic Vectors , Mesenchymal Stem Cells , Cell Biology , RNA, Messenger , Genetics , Metabolism , Transfection
6.
Journal of Experimental Hematology ; (6): 488-491, 2006.
Article in Chinese | WPRIM | ID: wpr-233561

ABSTRACT

The aim of this study was to evaluate the effect of dimethyl amiloride (DMA), a specific inhibitor of Na(+)/H(+) exchanger-1 (NHE-1), on intracellular pH value (pHi), proliferation and apoptosis of HL-60/ADM cells in vitro. After treatment with DMA at different doses, pHi of HL-60 and HL-60/ADM cell lines were determined by using pH-sensitive fluorescence dye BECEF-AM; the rate of growth inhibition of cells was detected with MTT assay; cell cycle was detected by flow cytometric DNA analysis; cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The results showed that pHi in HL-60/ADM cells was higher than that in HL-60 cells. After treatment with DMA at different doses, pHi decreased, the rate of growth inhibition and the rate of apoptotic cells in HL-60/ADM cells were all higher than those in HL-60 cells. Meanwhile, after treatment with DMA during 100 micromol/L to 150 micromol/L, the increase amplitude of G(0)/G(1) phase cells and the decrease amplitude of S + G(2)/M cells in HL-60/ADM cells were higher than those in HL-60 cells. It is concluded that by causing intracellular acidification, the NHE-1-specific inhibitor DMA inhibits proliferation of HL-60/ADM cells and induces apoptosis of HL-60/ADM cells, and the degree of this growth inhibition of HL-60/ADM cells is higher than that of HL-60 cells.


Subject(s)
Humans , Amiloride , Pharmacology , Apoptosis , Cell Cycle , Cell Proliferation , Doxorubicin , Pharmacology , Drug Resistance, Neoplasm , HL-60 Cells , Hydrogen-Ion Concentration , Sodium-Hydrogen Exchangers
7.
Journal of Experimental Hematology ; (6): 768-772, 2006.
Article in Chinese | WPRIM | ID: wpr-233499

ABSTRACT

The aim of this study was to evaluate the effect of all-trans retinoic acid (ATRA) on cell adhesion molecule expression and adhesion capacity of bone marrow stromal cells (BMSC) in patients after conditioning treatment for peripheral blood stem cell transplantation (PBSCT). BMSC of 27 patients before and after conditioning treatment for PBSCT were cultured in vitro. After treated with ATRA at 0.01, 0.1, or 1 micromol/L, expression of intercellular adhesion molecule-1 (ICAM-1) protein and vascular adhesion molecule-1 (VCAM-1) protein were detected by flow cytometry, and soluble ICAM-1 (sICAM-1) protein was determined by using radioimmunoassay. Then BMSC was co-cultured with CD34+ cells, and adhesion rate of BMSC to CD34+ cells was measured. The results showed that after pretreatment with conditioning regimen for PBSCT, the expressions of ICAM-1 and VCAM-1 proteins in BMSC and the expression level of sICAM-1 protein in supernatant of BMSC culture were down-regulated, and the adhesion rate of BMSC to CD34+ cells was decreased, after administration of ATRA, the expression of ICAM-1 protein in BMSC, sICAM-1 protein in culture medium and adhesion rate of BMSC to CD34+ cells all increased significantly, but expression of VCAM-1 protein changed no significantly. It is concluded that the ATRA can partly restore adhesion function of BMSC injured by pretreatment for PBSCT and contribute to hematopoietic reconstitution.


Subject(s)
Adolescent , Adult , Child , Humans , Middle Aged , Antigens, CD34 , Antineoplastic Agents , Pharmacology , Bone Marrow Cells , Metabolism , Pathology , Cell Adhesion , Cell Adhesion Molecules , Genetics , Coculture Techniques , Hematologic Neoplasms , Metabolism , Pathology , Therapeutics , Peripheral Blood Stem Cell Transplantation , Stromal Cells , Metabolism , Pathology , Tretinoin , Pharmacology , Tumor Cells, Cultured , Vascular Cell Adhesion Molecule-1 , Genetics
8.
Chinese Journal of Burns ; (6): 177-180, 2005.
Article in Chinese | WPRIM | ID: wpr-303670

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effects of burn serum on the erythropoiesis and granulopoiesis in bone marrow in mice, and to explore the possible underlying mechanism.</p><p><b>METHODS</b>Murine bone marrow cell (BMC) strain was prepared routinely and was employed in the establishment of the culture system of colony forming units of erythrocytes, or granulocytes and monocytes. To both sets of culture system normal murine serum (N group) and burn serum, which was collected from the mice with 15% full thickness burn at 12 postburn hours (PBH) and 1, 3, 5, 7 and 10 postburn days (PBD), (burn serum group) was added. In addition, positive control and blank control groups were set accordingly. The stimulating activity of all kinds of sera on the BMCs in the two sets of culture system was determined. The changes in the burn serum concentrations of EPO and GM-CSF were detected by radioimmunoassay, and the data were analyzed by logarithmic linear fitting correlation with the former influence of burn sera on the erythrocytes and granulocytes.</p><p><b>RESULTS</b>(1) Burn sera exhibited obvious stimulation promoting activity on the erythropoiesis and granulopoiesis in BMC, and the activity peaked (384 +/- 60 and 127 +/- 16 CFU) on 1 PBD and decreased thereafter to approach the values found in normal sera group (125 +/- 14 and 34 +/- 20 CFU) on 7 PBD. (2) The EPO content in burn serum was evidently higher than the normal value (P < 0.01) during 12 PBH to 7PBD period. The GM-CSF concentration was obviously higher than the normal value (P < 0.05) at 12 PBH and on 1 PBD. (3) The EPO concentration in burn serum was significantly and logarithmically correlated with the stimulation promoting activity of burn serum on erythropoiesis (r = 0.8570, P = 0.0137). But the GM-CSF concentration in culture with burn serum was not correlated with the stimulation promoting activity of burn serum on granulopoiesis (r = 0.7049, P > 0.05).</p><p><b>CONCLUSION</b>The sera harvested from burned mice during early postburn stage exhibited strong stimulation promoting activity on the erythropoiesis and granulopoiesis in bone marrow. The increased EPO level in burn serum might be the important factor contributing strong stimulation action on erythropoiesis, while increased GM-CSF level was not.</p>


Subject(s)
Animals , Mice , Bone Marrow , Metabolism , Burns , Blood , Therapeutics , Erythropoiesis , Erythropoietin , Metabolism , Granulocyte-Macrophage Colony-Stimulating Factor , Blood , Metabolism , Granulocytes , Hematopoietic Stem Cell Mobilization , Mice, Inbred Strains , Serum , Chemistry
9.
Journal of Experimental Hematology ; (6): 59-64, 2005.
Article in Chinese | WPRIM | ID: wpr-347826

ABSTRACT

To study the possibility of separation and culture of human umbilical cord blood adherent cell (HUCBAC), the umbilical cord blood CD34(+) cells were cultured in Dexter system in order to evaluate and observe the biological behavior of adherent cells in vitro. The results showed that all cells were cultured with Dexter system. By day 9-14 (at a median of 11.2 days), adherent cell colonies formed and reached their maximum at 15-22 days (mean 19.6 days), by day 28, all adherent cells spread over the bottom of Petri dish. By means of light microscopy, these cells were found to differentiate into three kinds of cells in culture of 28 days: fibroblast-liked cell, macrophage liked cell and small-round cells. The ratio of these three kinds of cells was 56.8%, 38%, 5.5% respectively. Cytochemistry assay revealed that the positive rate reached 100% in NSE stain and PAS stain; the adherent cell by ALP stain were shown 35% positive, but in POX stain the result was negative. Immunohistochemistry stain revealed that the positive rate of cord adherent cells for CD106, CD29, CD44, CD45, CD50, Fn, Ln, collagen IV etc reached 96%, 93%, 98%, 68%, 72%, 92%, 74%, 83% respectively. It is concluded there are hematopoietic adherent precursors in cord blood CD34(+) cells and the HUCBAC shows some biological behavior of hematopoietic stromal cells.


Subject(s)
Humans , Antigens, CD34 , Blood , Cell Adhesion , Allergy and Immunology , Cell Differentiation , Allergy and Immunology , Cells, Cultured , Fetal Blood , Cell Biology , Hematopoietic Stem Cells , Cell Biology , Allergy and Immunology , Hyaluronan Receptors , Blood , Immunohistochemistry , Integrin beta1 , Blood , Leukocyte Common Antigens , Blood , Vascular Cell Adhesion Molecule-1 , Blood
10.
Journal of Experimental Hematology ; (6): 269-273, 2005.
Article in Chinese | WPRIM | ID: wpr-356579

ABSTRACT

This study was aimed to explore the influence of anti-CXCR4 monoclonal antibody 12G5 on killing effect of cytosine arabinoside (Ara C) to HL-60 cell, and to assess its therapeutic value in marrow residual disease. HL-60 cells were cultured and co-cultured with leukemic stromal cells, and SDF-1 activity was inhibited with 10 microg/ml 12G5, then, killing effects of Ara C on HL-60 cells were investigated by MTT and morphology assay. Curves by MTT assay revealed that in the test group of 20 microg/ml Ara C, A(540) values decreased slowly but straightly, however, in control group A(540) values decreased markedly for the first two days, and increased from day 3 or 4. In the test group of 40 microg/ml Ara C, although increasing at constricted range of 7 - 9 days, A(540) values decreased in whole observing period of 12 days, while in control group A(540) values decreased markedly at day 0-3, and increased from day 4. Furthermore, two curves go across each other at day 5, and continue the increasing tendency. Morphology results showed that in both treated groups, the number of HL-60 cell decreased markedly and increased gradually in control group, but just contrary to test group. It is concluded that 12G5 may weaken the killing effect of Ara C on HL60 cell in earlier period, but reinforce the total killing effect and delay the occurrence of drug resistance simultaneously. Thus 12G5 has the therapeutic potential on marrow residual disease.


Subject(s)
Humans , Antibodies, Monoclonal , Pharmacology , Antimetabolites, Antineoplastic , Pharmacology , Cell Line, Tumor , Cell Survival , Cytarabine , Pharmacology , Dose-Response Relationship, Drug , Drug Synergism , HL-60 Cells , Receptors, CXCR4 , Allergy and Immunology
11.
Journal of Experimental Hematology ; (6): 274-277, 2005.
Article in Chinese | WPRIM | ID: wpr-356578

ABSTRACT

The aim was to analyze the expression level of stromal cell derived factor-1 (SDF-1) and its functional chemokine receptor CXCR4 in the patients with hematologic malignant tumor and their clinic significance. 28 patients with hematologic malignant tumor and 12 normal controls were chosen to be experimental objects. CXCR4 expressed on the cell membrane in bone marrow was enumerated by flow cytometry and serum level of SDF-1 was determined by ELISA assay. The result showed that the expression of SDF-1 and CXCR4 in hematologic malignant tumor were higher than that in normal controls, and the expression levels of two molecules were correlated. What is more, the different hematologic malignant tumor had different CXCR4 expression. In conclusion, the high expression of SDF-1 and CXCR4 in serum and bone marrow cells can be used as detective factors to hematologic malignant tumor. A correlation exists between the high expression of CXCR4 and the infiltration of hematologic malignant cells.


Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Chemokine CXCL12 , Blood , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Hematologic Neoplasms , Blood , Leukemia , Blood , Lymphoma , Blood , Receptors, CXCR4 , Blood
12.
Chinese Journal of Hematology ; (12): 421-424, 2005.
Article in Chinese | WPRIM | ID: wpr-255864

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of RNA interference inhibiting stromal cell derived factor-1 (SDF-1) expression on the proliferation and apoptosis of co-cultured Jurkat cells.</p><p><b>METHOD</b>Inhibition of SDF-1 expression by RNA interference (RNAi) was achieved by transferring SDF-1 specific short hairpin RNA (shRNA) expressing plasmid into cultured human acute leukemic bone marrow stromal cells. Resistant clones were obtained by G418 selection (group A). The concentration of SDF-1 protein in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The population double time (PDT), cell cycles, apoptosis rates and the expressions of PCNA, Bcl-2/Bax, Fas/FasL of co-cultured Jurkat cells were detected by cells counting, flow cytometry. TdT-mediated dUTP nick-end labelling (TUNEL) and immunocytochemistry (ICC), respectively. The un-transfected acute leukemic (group B) and normal (group C) bone marrow stromal cells were taken as controls.</p><p><b>RESULTS</b>The content of SDF-1 protein in supernatant of group A\[(384 +/- 41) pg/ml] was significantly lower than that in group B[(2474 +/- 271) pg/ml] or group C[(1324 +/- 154) pg/ml]. As group A compared with group B and group C, the PDT of co-cultured Jurkat cells was prolonged (group A: 42 h, vs group B: 29 h, group C: 33 h), and G(0)/G(1) stage cells increased [group A: (28.47 +/- 2.39)%, vs group B: (19.43 +/- 2.80)%, group C: (27.15 +/- 2.07)%], S stage cells decreased [group A: (25.57 +/- 1.90)%, vs group B: (74.48 +/- 3.23)%, group C: (60.99 +/- 2.33)%], G(2)/M stage cells increased [group A: (45.96 +/- 3.24)%, vs group B: (6.09 +/- 1.96)%, group C: (11.86 +/- 1.98)%], the apoptosis rate increased [group A: (15.2 +/- 0.8)%, vs group B: (5.4 +/- 0.7)%, group C: (9.5 +/- 0.4)%], and the expressions of PCNA, Bcl-2, Fas decreased; whereas the expressions of Bax and FasL were increased.</p><p><b>CONCLUSION</b>The inhibition of SDF-1 expression in bone marrow stromal cells inhibits the proliferation and promotes the apoptosis of co-cultured Jurkat cells.</p>


Subject(s)
Humans , Apoptosis , Genetics , Bone Marrow Cells , Metabolism , Cell Proliferation , Cells, Cultured , Chemokine CXCL12 , Genetics , Coculture Techniques , Gene Expression , Jurkat Cells , RNA Interference , Stromal Cells , Metabolism , Transfection
13.
Chinese Journal of Hematology ; (12): 458-460, 2005.
Article in Chinese | WPRIM | ID: wpr-255861

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effects of inhibiting stromal cell derived factor-1 (SDF-1) expression by RNA interference (RNAi) on adhesion and drug sensitivity of Jurkat cells co-cultured with bone marrow stromal cells.</p><p><b>METHODS</b>SDF-1 specific short hairpin RNA (shRNA) expressing plasmid was transferred into cultured human acute leukemic bone marrow stromal cells, positive clones were isolated by screening G418 resistance (Group A) , SDF-1 protein level in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The adhesion rates to bone marrow stromal cells layer and the drug sensitivity to doxorubicin of co-cultured Jurkat cells were detected by cell counting and MTT assay, respectively. The un-transfected bone marrow stromal cells of acute leukemia patient (Group B) or normal subject (Group C) were taken as control.</p><p><b>RESULTS</b>The level of secreted SDF-1 protein (pg/10(5) cells/week) in the supernatants of Group A, B and C were 1920 +/- 205, 12,370 +/- 1355 and 6620 +/- 770, respectively. Of co-cultured Jurkat cells in Group A, B and C, the adhesion rates after 24 h co-culturing were (28.8 +/- 2.6)%, (57.4 +/- 3.8)% and (45.2 +/- 4.0)%, respectively, and the IC50 values of doxorubicin were 585, 6162 and 1758 nmol/L, respectively.</p><p><b>CONCLUSION</b>Down-regulating SDF-1 expression of bone marrow stromal cells by RNAi reduces adhesion rates and enhances drug sensitivity to doxorubicin of their co-cultured Jurkat cells.</p>


Subject(s)
Humans , Bone Marrow Cells , Metabolism , Cell Adhesion , Cells, Cultured , Chemokine CXCL12 , Genetics , Metabolism , Coculture Techniques , Drug Resistance, Neoplasm , Gene Expression , Jurkat Cells , RNA Interference , Stromal Cells , Metabolism
14.
Journal of Experimental Hematology ; (6): 154-158, 2004.
Article in Chinese | WPRIM | ID: wpr-278777

ABSTRACT

This study was aimed to investigate the importance of chemokine SDF-1 in maintaining proliferation ability of acute myelocytic leukemia cell line HL-60 when the effects of SDF-1 on HL-60 cell proliferation were inhibited. Marrow stromal cells were cultured and co-cultured with HL-60 cells, and SDF-1 activity was blocked with anti-CXCR4 McAb. HL-60 cell activity was detected by MTT while cell cycle and the expression of CXCR4 on HL-60 cell membrane were observed by flow cytometry meanwhile. The internal calcium ionic concentration in HL-60 cell was detected as well before and after treated with 12G5. The results showed that 12G5 down-regulated the expression of CXCR4 on HL-60 cell membrane; HL-60 cells at G(0)/G(1) phase increased, but decreased at S phase; survive rate of leukemia cells reduced; the intercellular calcium ionic concentration of HL-60 cell decreased after treated with 12G5. It was concluded that brockage of the SDF-1 activity may inhibit proliferation of leukemia cell at certain level.


Subject(s)
Humans , Antibodies, Monoclonal , Pharmacology , Calcium , Metabolism , Cell Cycle , Cell Division , Cell Survival , Chemokine CXCL12 , Chemokines, CXC , Physiology , HL-60 Cells , Cell Biology , Receptors, CXCR4
15.
Journal of Experimental Hematology ; (6): 163-165, 2004.
Article in Chinese | WPRIM | ID: wpr-278775

ABSTRACT

To study the role of hematopoietic microenvironment abnormality in development of minimal residual disease and its mechanism, the viability of HL-60 cells was investigated by means of bone marrow stromal cell culture system or co-culture system of bone marrow stromal cell with HL-60 cells and idarubicin (IDA), flow cytometry and ELISA. The results showed that viability of HL-60 cells gradually decreased along with the increase of IDA dose and prolongation of culture time. Amount of HL-60 cells co-cultured with leukemia bone marrow stramal cells was significantly increased as compared with that of the control (P < 0.05). Bone marrow stromal cells or stromal cell conditioned medium reduced the effect of IDA on HL-60 cells in culture. In conclusion, leukemia bone marrow stromal cells contribute to increasing resistance of HL-60 cells to chemotherapeutic agents, and play some role in developing minimal residual disease.


Subject(s)
Humans , Bone Marrow Cells , Physiology , Cell Survival , Coculture Techniques , Dose-Response Relationship, Drug , Drug Resistance, Neoplasm , HL-60 Cells , Idarubicin , Pharmacology , Stromal Cells , Physiology
16.
Journal of Experimental Hematology ; (6): 265-269, 2004.
Article in Chinese | WPRIM | ID: wpr-352085

ABSTRACT

To explore the effects of bone marrow cells expanded under different conditions on hematopoietic reconstitution, in the liquid expanded cultural system with several cytokines and/or bone marrow stromal cell layers, the BMMNC of mice were expanded for 5 days. Then the expanded cells were transplanted into the lethal-dose irradiated mice via the caudal vein. The hematopoietic reconstitution of transplanted mice were evaluated by detecting the number of bone marrow nuclear cells and various colony forming cells. The results showed that ex vivo expansion of bone marrow mononuclear cells mediated with cytokines under cultural conditions could not improve the hematopoietic engraftment in post-irradiated mice, but the expansion supported by bone marrow stromal cells could benefit the reconstruction significantly regardless of addition with cytokines. In conclusion, the ex vivo hematopoietic cell expansion supported by bone marrow stromal cells can maintain the properties of the hematopoietic stem/progenitor cells for hematopoietic reconstitution.


Subject(s)
Animals , Female , Male , Mice , Bone Marrow Cells , Cell Biology , Bone Marrow Transplantation , Mortality , Hematopoiesis , Mice, Inbred BALB C , Stromal Cells , Physiology
17.
Journal of Experimental Hematology ; (6): 436-440, 2004.
Article in Chinese | WPRIM | ID: wpr-352047

ABSTRACT

To study the importance of chemokine SDF-1 in surviving of acute myelocytic leukemia cells HL-60, the adhesion ability of HL-60 and expression of Bcl-2, Fas protein when SDF-1 activity was blocked by anti-CXCR4 monoclonal antibody (12G5) were compared with those detected before MAb incubation, in this experiment, HL-60 cell were cultured and co-cultured with normal marrow stromal cell. The adhesion rate was detected while the expression of Bcl-2 and Fas proteins were assayed by immunohistochemical technique when SDF-1 activity was inhibited. The results showed that cell adhesion rate of HL-60 decreased while the expression Bcl-2 decreased and Fas increased. It is concluded that inhibition of SDF-1 activity increases cell apoptosis and thus reduces life-span of leukemia cell at certain level.


Subject(s)
Humans , Antibodies, Monoclonal , Pharmacology , Apoptosis , Cell Adhesion , Chemokine CXCL12 , Chemokines, CXC , Physiology , HL-60 Cells , Chemistry , Cell Biology , Proto-Oncogene Proteins c-bcl-2 , Receptors, CXCR4 , Physiology , fas Receptor
18.
Journal of Third Military Medical University ; (24): 544-546, 2001.
Article in Chinese | WPRIM | ID: wpr-737029

ABSTRACT

Objective To compare the irradiation-protective and inter-synergestic effects of E838,WR-2721, Rubia cordifolia, cystamin e hydrochloride and ethinyl estradiol on radiation and combined radiation-burn injury. Methods Above-mentioned drugs were given to the mice i ntraperitoneally, or intragastrcally, then, the mortality and the average surviv al d for 30 d were observed before and after the administration of the drug s. Results ①When drugs were before injury , the survival rate and the average survival d of the radiation and combined radiation-burn injured mice were increased obviously with the best effect in E838 and WR-2721. ②When drugs were given after injury, E838 and R. cordifolia also kept the effect. ③Combined appling WR-2721(pre) and E838(post)displayed a significant syner gistic reaction. Conclusion E838 and WR-2721 are more e ffective than the others in the prevention of radiation.

19.
Journal of Third Military Medical University ; (24): 544-546, 2001.
Article in Chinese | WPRIM | ID: wpr-735561

ABSTRACT

Objective To compare the irradiation-protective and inter-synergestic effects of E838,WR-2721, Rubia cordifolia, cystamin e hydrochloride and ethinyl estradiol on radiation and combined radiation-burn injury. Methods Above-mentioned drugs were given to the mice i ntraperitoneally, or intragastrcally, then, the mortality and the average surviv al d for 30 d were observed before and after the administration of the drug s. Results ①When drugs were before injury , the survival rate and the average survival d of the radiation and combined radiation-burn injured mice were increased obviously with the best effect in E838 and WR-2721. ②When drugs were given after injury, E838 and R. cordifolia also kept the effect. ③Combined appling WR-2721(pre) and E838(post)displayed a significant syner gistic reaction. Conclusion E838 and WR-2721 are more e ffective than the others in the prevention of radiation.

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