OBJECTIVETo investigate the expression of rat protamine (RP) gene in MEL cells and the effect on cell growth.

METHODSEukaryotic expression plasmid pCR-3.1-RP was constructed and transfected into MEL cells. The changes of cell growth rate, mitotic index and colony-forming rate in semi-solid medium were investigated.

RESULTSTransfected MEL cells showed lower growth rate, mitotic index and colony-forming rate. Volumes of cells were reduced and reduction of RNA transcription was observed.

CONCLUSIONThese results suggest that expression of RP in MEL cell may inhibit the cell growth and proliferation.

Animals ; Cell Division ; Leukemia, Erythroblastic, Acute ; genetics ; pathology ; Plasmids ; Protamines ; genetics ; metabolism ; Rats ; Transfection ; Tumor Cells, Cultured

Tanshinone II-A is a derivative of phenanthrene-quinone isolated from Salvia miltiorrhiza BUNGE, a traditional herbal medicine that is known to induce antiinflammatory, anti-oxidative and cytotoxic activity. We have examined cellular effects of Tanshione II-A on HL60 human promyelocytic leukemic cells and K562 human erythroleukemic cells. Tanshione II-A induced a dose- and time-dependent DNA fragmentation into the multiples of 180 bp and specific proteolytic cleavage of poly(ADP-ribose) polymerase in both cell lines. PI-staining and flow cytometry analysis of K562 cells following Tanshione II-A treatment showed an increase of the cells possessing hypodiploid DNA indicative of apoptotic state of cells. Caspase-3 activity was significantly increased during Tanshinone II-A treatment of both HL60 and K562 cells, whereas caspase-1 activity was not changed. These results suggest that Tanshione II-A induced HL60 and K562 cellular apoptosis that may be associated with the selective members of caspase family. Copyright 2000 Academic Press.
Antineoplastic Agents, Phytogenic/pharmacology* ; Antineoplastic Agents, Phytogenic/chemistry ; Apoptosis/physiology* ; Caspases/metabolism* ; Caspases/drug effects* ; Cell Cycle/drug effects ; DNA Fragmentation/drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal/pharmacology* ; Drugs, Chinese Herbal/chemistry ; Enzyme Activation/drug effects ; HL-60 Cells/pathology ; HL-60 Cells/metabolism ; HL-60 Cells/drug effects ; Human ; Lamiaceae/chemistry ; Leukemia/pathology* ; Leukemia/metabolism ; Leukemia/drug therapy ; Leukemia, Erythroblastic, Acute/pathology ; Leukemia, Erythroblastic, Acute/metabolism ; Leukemia, Erythroblastic, Acute/drug therapy ; Phenanthrenes/pharmacology* ; Phenanthrenes/chemistry ; Tumor Cells, Cultured

To compare the clinical outcome of autologous peripheral blood stem cell transplantation (APBSCT) and autologous bone marrow transplantation (ABMT) in treatment of patients with acute leukemia in first remission, 41 patients received APBSCT, 17 patients received unpurged ABMT and 30 patients received purged ABMT. The results showed that hematopoietic recovery was significantly earlier after APBSCT than that after purged or unpurged ABMT. The 3-year disease-free survival (DFS), relapse rate (RR) and transplant-related mortality (TRM) for all patients of 3 groups were 51.7%, 41.7% and 6.8%, respectively. DFS and RR were significantly influenced by disease types (ALL or AML) and intervals between diagnosis and CR(1) or CR(1) and transplant. The main causes of transplant-related death were infection and hemorrhage. After APBSCT, DFS, RR and TRM were 48.4%, 43.9% and 4.9%, respectively, and did not differ significantly from those found in unpurged ABMT (47.1%, 45.6% and 11.8%) or purged ABMT (66.5%, 29.6% and 6.7%). It is concluded that the clinical outcome of APBSCT is similar to unpurged or purged ABMT but APBSCT allows faster recovery of hematopoiesis and needs less transfusion support.
Acute Disease ; Adolescent ; Adult ; Bacterial Infections ; etiology ; mortality ; Bone Marrow Purging ; Bone Marrow Transplantation ; adverse effects ; Child ; Disease-Free Survival ; Female ; Follow-Up Studies ; Hemorrhage ; etiology ; mortality ; Humans ; Leukemia ; pathology ; therapy ; Leukemia, Erythroblastic, Acute ; pathology ; therapy ; Leukemia, Monocytic, Acute ; pathology ; therapy ; Leukemia, Myeloid, Acute ; pathology ; therapy ; Leukemia, Myelomonocytic, Acute ; pathology ; therapy ; Leukemia, Promyelocytic, Acute ; pathology ; therapy ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Peripheral Blood Stem Cell Transplantation ; adverse effects ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; pathology ; therapy ; Remission Induction ; Survival Rate ; Transplantation, Autologous

OBJECTIVESearching for more potent and less toxic HMBA-related agents.

METHODSHuman erythroleukemia cell K562, murine erythroleukemia cell (MEL) and its sub-line MEL DS19 were used as target cells to select a cell line which is the most sensitive to HMBA, then analyzed the activity of inducing differentiation of two new designed HMBA derivatives: HMBPA [hexamethylenebi (3-pyridin) amide] and Co-HDTA (ethylenediaminetetra acetic acid cobalt) using cell biology, cytochemical and molecular biology techniques.

RESULTSWe found that the MEL DS19 cells were most sensitive to HMBA (benzidine positive, B+ approximately 76%). Co-HDTA can inhibit the growth of MEL DS19, but induces differentiation just in a small population (B+ 2% approximately 4.5%). Between 0.02 approximately 5 micromol/L, HMBPA induces 3% approximately 8% cells committed to differentiation with little inhibition of cell proliferation. 1 micromol/L HMBPA and 2 mmol/L HMBA together, can obviously increase the percentage of differentiated cell (B+ approximately 72%), inhibit DNA synthesis and accelerate beta-globin transcription.

CONCLUSIONThe new HMBA derivatives may provide potential cancer differentiation inducers.

Acetamides ; chemistry ; pharmacology ; Animals ; Antineoplastic Agents ; pharmacology ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; Globins ; biosynthesis ; genetics ; Humans ; K562 Cells ; Leukemia, Erythroblastic, Acute ; metabolism ; pathology ; Mice ; Pyridines ; chemistry ; pharmacology ; RNA, Messenger ; genetics ; Tumor Cells, Cultured

The changes in the cellular main components of the mouse erythroleukemia cell line MEL for TFAR19 gene transfection were studied by the technology of Fourier transform infrared spectroscopy (FTIR). Using the method of gene transfection with liposome, we obtained MEL-TF19 cell line, which stably carries TFAR19, a novel apoptosis-related gene. The expression of the gene on mRNA level was confirmed by RT-PCR. Then, FTIR spectra of the cells were measured in the course of apoptosis induced by serum deprivation. Our results indicated that after being transfected with TFAR19 gene, MEL-TF19 cells exhibited relatively higher protein content, higher transcriptional activity, and relatively lower phospholipid content as compared with those exhibited by MEL cells. All the above changes reflect the apoptosis-promoting effect of TFAR19 gene, and maybe account for the cellular rheological changes after TFAR19 gene transfection, which were discovered in our previous study.
Animals ; Apoptosis ; genetics ; Apoptosis Regulatory Proteins ; Cell Line, Tumor ; Genes, Tumor Suppressor ; Leukemia, Erythroblastic, Acute ; genetics ; pathology ; Mice ; Molecular Sequence Data ; Neoplasm Proteins ; genetics ; pharmacology ; Spectroscopy, Fourier Transform Infrared ; Transfection

OBJECTIVETo explore the effects of Danshen Injection () on inhibition proliferation, inducing apoptosis and its possible mechanisms on human erythroid leukemic (HEL) cells.

METHODSThe commercial Chinese patent medicine of Danshen Injection was extracted and isolated from Chinese herb of Salvia miltiorrhiza bung. The inhibition effects of proliferation were assayed by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) method in HEL cells treated by Danshen Injection at various concentrations for 48 h. The cellular apoptosis was observed in morphology, analyzed by flow cytometry with annexin V and propidium iodide (PI) staining, and examined by DNA degradation ladder on agarose gel electrophoresis. Meanwhile, the expression levels of mutant Janus kinasez (JAK2) gene and phosphorylation-JAK2 (P-JAK2) protein were detected by allele specific-polymerase chain reaction and Western blot.

RESULTSThe proliferation of HEL cells was effectively inhibited by Danshen Injection in a dose-dependent manner, with suppression rates from 19.46±2.31% to 50.20±5.21%. Typical apoptosis cells was observed in Danshen Injection treated HEL cells, the rates of annexin V positive cells increased obviously in a dose-dependent manner, as well as the DNA degradation ladder of apoptosis revealed on gel electrophoresis. The expression levels of mutant JAK2 gene and P-JAK2 protein reduced gradually with increasing dosage of Danshen injection.

CONCLUSIONDanshen Injection could not only significantly inhibit the proliferation, but also induce apoptosis in HEL cells; down-regulation of the mutant JAK2 gene and P-JAK2 protein expressions are probably one of its molecular mechanisms.

Apoptosis ; drug effects ; Base Sequence ; Cell Proliferation ; drug effects ; DNA Primers ; Down-Regulation ; drug effects ; Humans ; Janus Kinase 2 ; genetics ; metabolism ; Leukemia, Erythroblastic, Acute ; enzymology ; metabolism ; pathology ; Mutation ; Phosphorylation ; Plant Extracts ; pharmacology ; Polymerase Chain Reaction ; Salvia miltiorrhiza ; chemistry

To investigate the relationship between nm23-H1 gene and human chronic myeloblastic leukemia we designed siRNAs which target nm23-H1 gene. According to the principles of designing siRNA, we selected three siRNAs and transfected them into K562 cells by lipofectamine2000. The expression levels of nm23-H1 mRNA were detected by reverse transcriptase polymerase chain reaction after transfection for 24 hours. The expression levels of nm23-H1 protein were assayed by immunocytochemical method after transfection for 48 hours. And after transfection for 24, 48 and 72 hours, cell proliferation was determined by MTT method. Among the three siRNAs, siNM526 can effectively inhibit the expression of nm23-H1 on mRNA and protein levels. The growth of K562 cells was suppressed after transfection of siNM526. These results suggest that low expression level of nm23-H1 in K562 cells inhibited cell proliferation, namely reduced malignant degree of them. Therefore nm23-H1 gene might be a potential target of leukemia treatment.
Cell Proliferation ; Humans ; K562 Cells ; Leukemia, Erythroblastic, Acute ; genetics ; pathology ; NM23 Nucleoside Diphosphate Kinases ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo observe matrine-induced erythroid differentiation of K562 cells in relation to activation of the apoptotic pathway in vitro.

METHODSK562 cells were cultured in the presence or absence of matrine at different concentrations for 4 days, and the morphological and ultramicrostructural changes of the cells were observed using inverted microscopy and transmission electron microscopy, respectively. The expression of apoptosis-related protein p27kip1 was detected by immunocytochemistry.

RESULTSCompared to untreated K562 cells, the cells treated with matrine at 0.10 g/L exhibited apoptostic characteristics in the cellular morphology and ultramicrostructure, with the expression of p27kip1 protein upregulated in a concentration- and time-dependent manner.

CONCLUSIONMatrine-induced erythroid differentiation of K562 cells is associated with cell apoptosis, and upregulation of p27kip1 protein expression may play a crucial role in the process of apoptosis.

Alkaloids ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; physiology ; Cyclin-Dependent Kinase Inhibitor p27 ; biosynthesis ; Dose-Response Relationship, Drug ; Humans ; Immunohistochemistry ; K562 Cells ; Leukemia, Erythroblastic, Acute ; metabolism ; pathology ; Microscopy, Electron, Transmission ; Quinolizines ; pharmacology ; Signal Transduction ; drug effects ; Time Factors

Using transplantable erythroblastic leukemia cells of EL9611(H-2d), the cells were inoculated to CB6F(1)(H-2d/b) generation of BALB/c x C57BL/6 mouse, the biological characterization of erythroblastic leukemia in haploidentical mouse was studied, that provides an experimental model for the study of graft-versus leukemia (GVL) with bone marrow or stem cell transplantation. When 10(3) - 10(8) of the spleen cells of EL9611(H-2d) had been intravenously inoculated to CB6F(1) mouse, the erythroblastic leukemia cells were transplanted successively and the F(1) generation of erythroblastic leukemia model in mice was established with 100% incidence of erythroblastic leukemia. There was a linear relationship between the survival time and the number of leukemic cell. The survival time of the mice was (9.6 +/- 0.8) days when 10(6) cells were inoculated. If the CB6F(1) mouse was transplanted successively for four generations, the incidence was 100%. The main targets for the leukemic EL9611(H-2d) cells were liver, spleen and marrow. The reaction of the erythroblastic leukemia cells for hemoglobin staining was positive, while the peroxidase reaction was negative. These cells were sensitive to some chemotherapeutic drugs, such as cytosine arabinoside and cyclophosphamide. This study presents the convenience for the studies on the GVL with haplo-allogeneic transplantation, in the F(1) generation of erythroblastic leukemia model of the commonly-used CD57BL/6 x BALB/c mouse.
Animals ; Cell Division ; Disease Models, Animal ; H-2 Antigens ; analysis ; Leukemia, Erythroblastic, Acute ; immunology ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Neoplasm Transplantation ; Survival Analysis ; Tumor Cells, Cultured

AML relapsing as ALL has rarely been reported. We describe the case of a 62-yr-old man who was diagnosed with erythroleukemia with a complex karyotype and achieved complete hematologic and cytogenetic remission after induction chemotherapy. However, 4 months after the initial diagnosis, he showed relapse with blasts showing a different morphology and immunophenotype and was diagnosed with precursor B-cell ALL. The relapsing precursor B-cell ALL presented with the same leukemic clones as the primary erythroleukemia. Cytogenetic analysis of his bone marrow (BM) at the time of the primary erythroleukemia showed complex karyotypic abnormalities, including monosomy 5 and monosomy 7. At relapse, his BM showed reemergence of these leukemic clones of complex karyotypic abnormalities with clonal switch. To our knowledge, this is the first case of a lineage switch from erythroleukemia to ALL.
Acute Disease ; Antimetabolites, Antineoplastic/therapeutic use ; Bone Marrow Cells/pathology ; Cell Lineage ; Cell Transformation, Neoplastic ; Chromosome Deletion ; Chromosomes, Human, Pair 5 ; Chromosomes, Human, Pair 7 ; Cytarabine/therapeutic use ; Drug Therapy, Combination ; Humans ; Immunophenotyping ; Karyotyping ; Leukemia, Erythroblastic, Acute/*diagnosis/drug therapy ; Male ; Middle Aged ; Monosomy ; Naphthacenes/therapeutic use ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma/*diagnosis/pathology ; Recurrence