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OBJECTIVE: To investigate the protection of hepatocyte growth factor (HGF) on CML cell line K562 from apoptosis induced by etoposide (VP-16) and its molecular mechanism. METHODS: Quantitative and qualitative analyses on cell morphological change of apoptosis were performed through acridine orange (AO) staining and HE staining, and fluorescent flow cytometry.The test analyzes membrane on the surface of the PS evagination and integrity of cell membrane surface and mitochondrial membrane potential changes were performed through Annexin V-FITC/PI double dyeing and JC-1 cell dyeing tests, and apoptotic factors such as Bcl-2, Bax, Caspase-3 and Caspase-9 were measured by SYBR Green (Takara) qRT-PCR. RESULTS: The HE and AO staining revealed that apoptotic rates in HGF+VP-16 groups were significantly lower than those in VP-16 groups (P<0.05, P<0.05), HGF can inhibit the apoptosis of cells induced by VP-16; FCM (Annexin V-FITC/ PI and JC-1) tests showed that cells apoptotic rates in HGF+VP-16 groups were significantly lower than those in VP-16 groups (P<0.05, P<0.001), indicating that HGF has the anti-apoptosis function. Apoptosis related gene mRNA expression tests found that the Bcl-2 mRNA expression in HGF+VP group was obviously higher than that in the VP-16 group (P<0.001), while Bax mRNA, Caspase-3 mRNA, and Caspase-9 mRNA expressions were significantly lower than those in the VP-16 group (P<0.05, P<0.001, P<0.001), suggesting that HGF possesses antiapoptotic effect through inhibiting apoptosis gene expression and promoting the antiapoptotic gene expression simultaneously. CONCLUSION: HGF can significantly protect K562 cells from apoptosis induced by VP-16 through the HGF/c-Met way to regulate PI3K/AKT pathway.
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OBJECTIVE: To investigate the inhibitory activity, induced differentiation-inducing activity and apoptosis-inducingactivity of hydroxyl morpholine (QDML-01) on chronic myelocytic leukemia cells line K562. METHODS: The cell growth curve was drawn based on cell counting method. The IC50 value of QDML-01 and positive control medicine to K562 cells were evaluated by methyl thiazolyl tetrazolium (MTT) assay method. Double soft agar assay method was carried out to study the ability of cell proliferation to determine efficacy of phamacognosy. The pathomorphism was analyed by the Wright-Giemsa staining method. The mechanism of cell apoptosis from morphology and gene level were investigated, by AO-EB double-staining method and DNA breakage test. The effect of QDML-01 on K562 cells from the protein level was determined by Western-blot. RESULTS: The growth curves showed the K562 cells had strong cell vitality. They came into logarithmic phase on the third generation. The MTT assay results showed that the IC50 values of QDML-01 and imatinib to K562 cells were 5. 81 and 596.88 nmol ·L-1. Double soft agar colony formation test showed that clone formed at 21 d and the inhibitory rate of QDML-01 was 81.7%. It indicated that K562 cells were sensitive to QDML-01. Morphology test result showed that QDML-01 induced K562 cells to normal cells. The results of AO-EB double-staining method showed that QDML-01 induced the apoptosis of K562 cells. The study of DNA breakage test indicated that QDML-01 can induce the apoptosis of K562 cells to produce DNA banding with step-like. Western-blot analysis result suggested that QDML-01 can downregulated the expression of P210bcr/abl protein. CONCLUSION: QDML-01 has the inhibitory activity on chronic myelocytic leukemia cells line K562 by promoting the apoptosis of K562 cells and inducing differentiation to normal cells.
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Objective Research of chromosome’s influence and curative effect on first and second generation tyrosine kinase inhibitor therapy to CML patients.Methods Giving conventional genetic analysis to 80 Ph+ CML patients,and contrast CML patients’chromosome changing situation with first and second generation tyrosine kinase inhibitor therapy.Results There were 1 1 cases with other abnormalities of chromosome number and structure in 80 cases of Ph+ CML patients,and 10 cases were resistant or intolerant to imatinib.35 patients (87.5%)achieve sustained complete cytogenetic remission (CCyR)who treated with imatinib (TKI-Ⅰ)in the total 40 cases,in these 35 patients,7 cases (17.5%)got CCyR in 3 months;10 cases (25%)got CCyR in 6 months,13 cases (32.5%)got CCyR in 12 months,and 5 cases (12.5%)got CCyR in 18 months.33 patients (82.5%)achieve sustained completecytogenetic remission (CCyR)who treated with dasatinib/nilotinib (TKI-Ⅱ)in the total 40 cases,in these 33 cases,16 cases (40%)got CCyR in 3 months;9 cases (22.5%)got CCyR in 6 months,5 cases (1 2.5%)got CCyR in 1 2 months,and 3 cases (7.5%)got CCyR in 1 8 months.Conclusion Ph+ CML patients combined with other chromosome abnormity were more easily to be resistant or intolerant to imatinib,espe-cially in acceleratd phase and blastic phase.First and second generation tyrosine kinase inhibitor have little difference to treat with CML patients in long time efficacy,but the second generation effect is better than first generation in short time effica-cy.
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Objective To discuss the observation and nursing of adverse reactions in chronic myelocytic leukemia patients receiving imatinib therapy.Methods Adverse reactions were observed and recorded in 193 chronic phase myelocytic leukemia patients who received imatinib therapy,and corresponding treatment and nursing were given to them.Results Among 193 patients,more than 60％ of patients had adverse reactions,of which,54％ of patients showed gastrointestinal adverse reactions including nausea,vomiting,anepithymia and diarrhea; 22％ of them had muscle and bone pain; 7％ had rash; 65％ got edema.After proper treatment and nursing,all adverse effects obtained satisfactory remission.Condusions During the treatment course of chronic myelocytic leukemia patients using imatinib,careful observation of any possible adverse reactions,and giving corresponding treatment and nursing can facilitate good compliance and longterm remission of patients.
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Objective To increase the sensitivity of residual leukemic cells detectin in chronic myelocytic leukemia(CML) patients with RT-PCR,the optional annealing temperature and PCR cycles were studied to confirm bcr-abl fused gene types,and bcr-abl mRNA transcripts were monitored by RQ-PCR to study the relation with CML at different phases. Methods Through changing the PCR conditions, the annealing temperature was measured from 55℃ to 60℃, and the number of reaction cycles was increased from 30 to 45.All 22 samples were examined, and bcr-abl mRNA transcripts were quantified by RQ-PCR kit. Results Bcr-abl fused gene types were found in 22 samples,of all 9 cases were b_2a_2 type, 13 cases were b_3a_2.When the annealing temperature was set for 58℃ and the number of reation cycles was set for 45,10~3 copies/ul standard samples was detected.18 samples were positive tested by RQ-PCR kit,and the value was between 10~2 to 10~6 copies/g.There were significant differce between the results of chronic phase samples and those of accelerated phase. Conclusions The RT-PCR is a reliable,sensitive and reproducible method of monitoring CML patients.The real-time RT-PCR is useful in evaluating leukemic burden,assessing response to treatment and predicating the prognosis of the disease.
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Purpose:To study the secretion and gene expression of Angiogenesis factors in the patients with CML and study the effect of Angiogenesis in the occurrence and development of CML. Methods:Concentration of VEGF in peripheral blood was determined by using ELISA. Myeloid tissue was extracted from all CML cases and ITP patients to detect MVD by using CD34 labelling. At the same time the level of VEGF,b-FGF were detected by using RT-PCR in both peripheral blood and myeloid cells.Results:Our results showed that the concentration of VEGF was obviously higher in the peripheral blood of CML patients(177.53?153.45 pg/ml) than in those of control group(73.12?19.82 pg/ml). The gene expression of VEGF and b-FGF were both higher than those of control group. The difference has statistical significance(P
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We report a case of prostatic abscess in a 46-year old man with chronic myelocytic leukemia. Preoperative transrectal ultrasonography and computerized tomography confirmed the diagnosis of prostatic abscess, which was treated with pus drainage via transurethral resection of prostate and broad-spectrum antibiotics.
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Humans , Middle Aged , Abscess , Anti-Bacterial Agents , Diagnosis , Drainage , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Suppuration , Transurethral Resection of Prostate , UltrasonographyABSTRACT
Object To investigate the effect of Angelica polysaccharide (APS) on the induction of chronic myelocytic leukemia cells into chronic myelocytic leukaemia dendritic cells (CML-DCs). Methods Bone marrow monocytes from CML patients were cultured in GM-CSF/IL-4 or in GM-CSF/IL-4 combined with APS in each concentration (50, 100, 200 mg/L), respectively. The morphotype of CML-DCs was identified by optical microscope or electron microscope, CML-DCs viability was calculated by Trypan Blue exclusion. The phenotype of CML-DCs (CD 80, CD 86, and CD 83) was identified by flow cytometry. The capability of stimulating auto-lymphocyte or allo-lymphocyte proliferation was tested with mixed leukocyte reaction (MLR). Results Bone marrow monocytes from CML patients, which were cultured in GM-CSF/IL-4 or in GM-CSF/IL-4/APS showed typical morphotype and expressed the high level phenotype of CML-DCs. The capability of proliferation and the survival rate of CML-DCs were enhanced markedly and the expression of CD 83, CD 80, and CD 86 on CML-DCs were significantly increased when CML-DCs were cultured in GM-CSF/IL-4/APS. The capability of stimulating lymphocyte proliferation was more competent in 100 mg/L APS group. Conclusion The expression of CD 83, CD 80, and CD 86 on CML-DCs cultured in GM-CSF/IL-4/APS is significantly higher than those in GM-CSF/IL-4. The capability of CML-DCs of stimulating lymphocyte proliferation is more potential in GM-CSF/IL-4/APS than in GM-CSF/IL-4. APS can promote the induction and mature of CML-DCs cultured in IL-4 and GM-CSF.
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The leukemic cellular surface antigens of 45 cases for ANLL and 36 cases of CML were detected and classified with a series of the domestic McAbs by immunoflurescent technique. The immunophenotype of ANLL was compared with its cellular morphologic classification. The difference of the surface antigens between the CML instable phase and the CML in blast crisis was analyzed. Four cases of acute leukemias (AL) which could not be classified morphologically were classified by McAbs. FAB M_1～M_5 could be divided into 3 phenotype groups: myeloblastic, promylocytic and promoncytic phenotype. The immunophenotype of ANLL tended to agree with its cellular morphologic classification. All 28 cases of CML in stable phase reacted with a neutrophil-McAb. However, there was no this antigen expression in 8 cases of CML in blast crisis. The immunophenotype of 4 cases of AL could be made with McAbs . Of these 4 cases. 3 were AL and 1 ANLL.