1.Mechanism of cinnamic aldehyde-inducing apoptosis of chronic myeloid leukemic cells in vitro.
Li-Qiong LIU ; Ze-Lin LIU ; Xin WANG ; Hai-Yan CUI ; Meng-Di JIN ; Dan-Yu WANG ; Shi-Ang HUANG
Journal of Experimental Hematology 2011;19(3):617-620
The aim of this study was to investigate the apoptosis-inducing effect of cinnamic aldehyde (CA) on chronic myeloid leukemic (CML) cells and its mechanism. K562 cells and primary bone marrow mononuclear cells (MNC) from patients with CML were treated by various concentrations of CA. Flow cytometry was employed to measure the apoptosis of K562 cells and primary CML bone marrow MNC. Western blot was used to determine the expression of C-MYC and the phosphorylation of CrkL in K562 cells, and real-time polymerase chain reaction (real-time PCR) was used to quantify the expression of BCR-ABL mRNA in K562 cells. The results indicated that CA induced the apoptosis of K562 cells in a time- and dose-dependent manner. CA induced apoptosis of CML MNC dose-dependently. CA inhibited the expression of BCR-ABL mRNA and C-MYC, reduced CrkL phosphorylation levels in K562 cells. It is concluded that CA induces apoptosis of CML cells in vitro. Down-regulation of the expression and function of BCR-ABL may be one of its most important anti-leukemia mechanisms.
Acrolein
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analogs & derivatives
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pharmacology
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Apoptosis
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drug effects
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Fusion Proteins, bcr-abl
;
metabolism
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Gene Expression Regulation, Leukemic
;
Humans
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K562 Cells
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Leukemia, Myelogenous, Chronic, BCR-ABL Positive
;
metabolism
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pathology
2.Research Advance on the Effect of Autophagy on the Survival of Chronic Myeloid Leukemia Cells--Review.
Yang YANG ; Hui-Jun CHEN ; Jian LI
Journal of Experimental Hematology 2020;28(6):2093-2096
Tyrosine kinase inhibitor (TKI) has significantly improved the treatment of chronic myeloid leukemia (CML), however the resistance often resulted in treatment failure. Currently, it is known that the survival of CML cells can be affected by regulating autophagy, oxidative stress and mitochondrial metabolism, among which autophagy is an evolution-conserved catabolism process, and closely related to the pathogenesis of CML, thus playing a dual role in regulating the biological characteristics of cells. On the one hand, autophagy can promote the apoptosis of CML cells, and also can induce the drug resistance of CML cells on the other hand. In this review, the effect of autophagy on CML cells was summarzed briefly, so as to provide a useful idea to explore the combination of TKI with the autophagy inhibitor or inducer to overcome the resistance of CML to TKI.
Apoptosis
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Autophagy
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Drug Resistance, Neoplasm
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Fusion Proteins, bcr-abl
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Humans
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Leukemia, Myelogenous, Chronic, BCR-ABL Positive
;
Protein Kinase Inhibitors/pharmacology*
;
Research
3.Recent Advance of Newly Therapy for Chronic Myeloid Leukemia with BCR-ABLT315I Mutation--Review.
Hu-Rong LAI ; Qian-Miao WU ; Ya-Zhi YANG ; Jian LI
Journal of Experimental Hematology 2023;31(5):1579-1583
BCR-ABLT315I mutation is the main mechanism of resistance to the first and second generation tyrosine kinase inhibitor (TKI) for patients with chronic myeloid leukemia (CML). Ponatinib as the third generation TKI has been found that can significantly improve the prognosis of CML patients with T315I mutation. However, the latest report has discovered that the T315I compound mutant is even resistant to ponatinib, which aroused the enthusiasm of research on the mechanism of CML resistance and targeted therapy once again. Previous studies have shown that TKI combined with other targeted drugs is effective to CML patients with drug resistance or relapse due to T315I mutation. The latest research has found that the allosteric inhibitor asciminib combined with TKI therapy is equally effective to CML patients with T315I compound mutant, but the specific mechanism is not yet clarified. This review will focus on the latest research progress of therapy for CML with BCR-ABLT315I mutation, hoping to provide reference for researching new drugs and improve therapy for treating CML with T315I mutation.
Humans
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Drug Resistance, Neoplasm/genetics*
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Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics*
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Fusion Proteins, bcr-abl/genetics*
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Protein Kinase Inhibitors/therapeutic use*
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Mutation
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Antineoplastic Agents/pharmacology*
4.The effect of multi-unit ribozymes on the growth inhibition and apoptosis induction of CML K562 cells.
Qi FENG ; Bingzhong SUN ; Kai SUN ; Zhenchuan SHANG ; Sha WANG ; Wei WANG ; Yongtong ZHAO ; Zhen YAN ; Wei HAN ; Yingqi ZHANG
Chinese Journal of Oncology 2002;24(5):435-439
OBJECTIVETo investigate the possibility of multi-unit ribozymes to purge bone marrow of chronic myelocytic leukemia (CML), its in vitro cleavage ability and the reversal effect on CML cell's malignant phenotype.
METHODSAs bcr-abl fusion gene plays an important role in CML pathology, three single-unit ribozymes were designed and synthesized in 44 base pairs near the fusion point, two enzyme cleavage sites on bcr gene and one on abl gene. Multi-unit ribozymes' in vitro transcription and retroviral vector through gene recombination were constructed. Then, its in vitro cleavage ability was tested and the retroviral vector was transfected into K562 cell. Through MTT assay, the incorporation rate of (3)H-TdR, RT-PCR, Southern and Northern blot hybridization, flow cytometry, transmission and scanning electron microscopy were used to study the effect of multi-unit ribozymes on CML cell proliferation, cell structure, cell cycle and the induction of apoptosis.
RESULTSMulti-unit ribozymes had in vitro cleavage efficiency of 70.8%. After the transfection of multi-unit ribozymes retroviral vector into K562 cell, cell proliferation and DNA synthesis were greatly reduced with an inhibition rate of about 50% after 96 hours of transfection. Multi-unit ribozymes could cleave K562 cell's RNA with a reduction rate about one 1 000 th of the original. By flow cytometry (FCM), 18.4% cells underwent apoptosis after 72 hours transfection with most of the cells blocked in the G phase. Here, the ratio in S phase was lowered by 41.9%. Under transmission and scanning electron microscope, compaction of nuclear chromation and apoptosis bodies were observed in the transfected cells.
CONCLUSIONMulti-unit ribozymes possess high cleavage ability in vitro. The ribozymes, whose retroviral vector being transfected into CML cell, are able to express a lasting ability to cleave the fusion gene, induce apoptosis, reduce cell proliferation, revert the malignant phenotype. It is possible to make use of multi-unit ribozymes to purge CML bone marrow. Therefore, multi-unit ribozymes may very well be valuable in the gene therapy of CML.
Apoptosis ; Cell Division ; drug effects ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Humans ; K562 Cells ; RNA, Catalytic ; metabolism ; pharmacology
5.Effect of dihydroartemisinin on the expression of BCR/ABL fusion gene in leukemia K562 cells.
Jia-liang GAO ; Xian-ping DING ; Qi-jie LI ; Zeng-liang XIA ; Qing-jie XIA
Chinese Journal of Medical Genetics 2012;29(1):19-22
OBJECTIVETo investigate the effect of dihydroartemisinin (DHA) on the BCR/ABL fusion gene in leukemia K562 cell.
METHODSK562 cells were cultured in vitro. The rate of proliferation inhibition of cells treated with various concentrations of DHA were determined by using [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) method. Expression of BCR/ABL fusion gene was analyzed by reverse transcription(RT-PCR) before and after DHA treatment. Apoptosis of K562 cells was detected by flow cytometry.
RESULTSThe growth of K562 cells was inhibited when the concentrations of DHA were 10-160 umol/L. With the added dose of DHA, the growth inhibition was remarkable, with the rate of inhibition risen from 52.76% to 94.65%. The expression of BCR/ABL fusion gene, as detected by RT-PCR after incubating the K562 cells with 20 umol/L DHA, measured as ΔCt = 4.45 ± 0.25 after 12 h and ΔCt = 5.23 ± 0.21 after 24 h, which was significantly lower compared with that of the control ( ΔCt = 4.23 ± 0.21, P < 0.05).
CONCLUSIONDHA can inhibit the proliferation of leukemia K562 cells and facilitate the induction of apoptosis by downregulating the expression of BCR/ABL fusion gene.
Artemisinins ; pharmacology ; Fusion Proteins, bcr-abl ; biosynthesis ; genetics ; Gene Expression ; drug effects ; Genes, abl ; drug effects ; Humans ; K562 Cells ; Leukemia ; genetics ; Tumor Cells, Cultured
6.Effects of POH in combination with STI571 on the proliferation and apoptosis of K562 cells.
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(1):41-4
The effects of monoterpene perilly alcohol (POH) alone or in combination with STI571 on the proliferation and apoptosis of the cell line K562 positive for Bcr/Abl were investigated. By using cell culture, the effect of the drugs on the proliferation of the cells was studied. TUNEL and flow cytometry assay of FITC-Annexin V and PI labeled cells were applied to detect the effects of the drugs on the apoptosis of the cells. The results showed that at 36 h, IC50 of POH on K562 positive for Bcr/Abl and HL-60 negative for Bcr/Abl were 81.0 +/- 11.3 micromol/L and 113.6 +/- 23.4 micromol/L respectively (P>0.05). POH could inhibit the proliferation of K562 in a time- and dose-dependent manner with the inhibitory rate of 100 micromol/L POH on K562 cells at 36 h being (53.2 +/- 3.65)%. K562 cells were more sensitive to STI571 than POH. IC50 of STI571 on K562 cells in 36 h was (0.256 +/- 0.054) micromol/L. In a time- and dose-dependent manner, POH induced the apoptosis of K562 cells with the percentage of apoptotic cells by 100 micromol/L POH at 40 h being (21.0 +/- 3.3)%. Both 100 micromo/L POH and 0.2 micromol/L STI571 had the same inhibitory effects on the K562 cells at 36 h. But at 12 and 24 h, the inhibitory rate of POH was significantly higher than that of STI571 (P<0.05) and the ability of STI571 inducing apoptosis at 36 h was greater than that of POH. 50 micromol/L, 100 micromol/L and 200 micromol/L POH in combination with 0.2 micromol/L STI571 could obviously increase the inhibitory effects on the cellular proliferation. Combined use of 50 micromol/L, 100 micromol/L, 200 micromol/L with 0.2 micromol/L STI571 could strongly induced apoptosis, especially 200 micromol/L POH in combination with 0.2 micromol/L STI571. It was concluded that the antileukemia effect of POH had no obvious Bcr/Abl positive selectivity. POH can inhibit the proliferation of K562 and induce the apoptosis in a time- and dose-dependent manner. K562 cells were more sensitive to STI571 than POH. POH in combination with STI571 could obviously enhance the abilities of STI571 inhibiting the proliferation and inducing apoptosis of K562 cells.
Antineoplastic Agents/*pharmacology
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Apoptosis/*drug effects
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Dose-Response Relationship, Drug
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Drug Synergism
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Fusion Proteins, bcr-abl/analysis
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HL-60 Cells
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K562 Cells
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Monoterpenes/*pharmacology
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Piperazines/pharmacology
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Pyrimidines/*pharmacology
7.Clonal expansion of TCR Vbeta subfamily T cells induced by bcr3-abl2 peptide.
Yu-ping ZHANG ; Yang-qiu LI ; Li-jian YANG ; Shao-hua CHEN ; Xue-li ZHANG ; Zhen WANG ; Xiu-li WU ; Geng-xin LUO
Chinese Journal of Hematology 2004;25(2):95-99
OBJECTIVETo investigate the clonal expansion of T cell receptor (TCR) Vbeta subfamily T cells from cord blood induced by bcr3-abl2 peptide in vitro.
METHODST cells from 3 units of cord blood were amplified by anti-CD(3) monoclonal antibody (McAb) and IL-2 with or without synthetic b3a2 peptide. T cell specific cytotoxicity was analyzed by lactate dehydrogenase (LDH) assay, TCR Vbeta subfamilies by using reverse transcriptase-polymerase chain reaction (RT-PCR) and genescan technique.
RESULTSbcr3-abl2 peptide specific cytotoxicity T cells were successfully induced from the 3 units of cord blood by synthetic b3a2 peptide. Compared with that in CD(3) McAb induced cells, distribution pattern of TCR Vbeta repertoire was different in T cells induced with b3a2 peptide. Oligoclonal and oligoclonal tendency TCR Vbeta subfamily T cells could be identified in cord blood T cells induced by b3a2 peptide in 1 or 2 weeks, whereas those induced by anti-CD(3) McAb and IL-2 were mostly polyclonal.
CONCLUSIONThe cytotoxicity T cells with anti-CML specificity could be induced by b3a2 peptide. The specific anti-CML cytotoxicity may be derived from the clonal expansion TCR Vbeta subfamily T cells.
Antibodies, Monoclonal ; immunology ; CD3 Complex ; immunology ; Fusion Proteins, bcr-abl ; pharmacology ; Genes, T-Cell Receptor beta ; Humans ; Interleukin-2 ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; T-Lymphocytes, Cytotoxic ; immunology
8.Effect of A Novel Emodin Derivative on Chronic Myelogenous Leukemia K562 Cells and Imatinib-resistant K562/G01 Cells.
Bo-Jun LI ; Ting-Bo LIU ; Wen-Feng WANG ; Min-Hui LIN ; Jian-Da HU
Journal of Experimental Hematology 2016;24(1):1-7
OBJECTIVETo explore the effect of a novel emodin derivative E19 on proliferation inhibition and apoptosis induction of human chronic myelogenous leukemia (CML) cell line K562 and imatinib-resistant CML cell line (K562/G01), and to clarify the involved mechanisms.
METHODSMTT and colony formation test were used to detect the cell proliferation. Apoptotic induction effects were examined by DAPI staining method and DNA ladder assay. Western blot was performed to detect the changes of P210(Bcr-Abl) protein.
RESULTSThe emodin derivative E19 could efficiently inhibit proliferation and induce apoptosis in K562 and K562/G01 cells. IC50 of K562 cells and IC50 of K562/G01 cells were (1.20 ± 0.19) µmol/L and (1.22 ± 0.16) µmol/L, respectively. DNA fragmentation in K562 cells and K562/G01 cells confirmed that the E19 induced apoptosis in dose-dependent manner. Western blot showed that emodin derivative inhibited phosphorylation of P210 protein in K562 cells and K562/G01 cells and down-regulated the expression level of P210 in dose- and time-dependent manners.
CONCLUSIONThe emodin derivative E19 can efficiently inhibit growth and induce apoptosis of K562 cells and K562/G01 cells, while the inhibition of phosphorylation of P210 protein and down-regulation of P210 protein expression may be involved in these processes.
Apoptosis ; drug effects ; Cell Proliferation ; Down-Regulation ; Drug Resistance, Neoplasm ; Emodin ; analogs & derivatives ; pharmacology ; Fusion Proteins, bcr-abl ; metabolism ; Humans ; Imatinib Mesylate ; pharmacology ; K562 Cells ; drug effects ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; pathology ; Phosphorylation
9.Preliminary study on the molecular mechanism of K562 cell apoptosis induced by As2S2.
Jun-e LI ; Guan-lin SUN ; Ying-li WU ; Wei-li WU
Chinese Journal of Oncology 2003;25(3):220-224
OBJECTIVETo investigate the apoptotic inducing effect of As(2)S(2) on K562 cells.
METHODSThe apoptotic inducing effect of As(2)S(2) on K562 cells was determined by flow cytometry, DNA fragmentation analysis and morphology observation. Expression of protein was determined by Western-blot. RT-PCR was used to evaluate changes in gene expression.
RESULTSApoptosis of K562 cells was induced by 48 - 72 h exposure to 5 micromol/L As(2)S(2). Apoptosis was induced in (34.4 +/- 3.3)% treated cells by 72 h exposure to 3 micro mol/L As(2)S(2), in (21.8 +/- 3.6)% treated cells by 48 h exposure to 5 micromol/L As(2)S(2) and in (46.0 +/- 5.2)% treated cells by 72 h exposure to As(2)S(2) at the same concentration. With 5 micromol/L As(2)S(2), the protein level of Bcr-Abl and JAK2 decreased, while bax expression was upregulated and c-myc was downregulated both in protein and mRNA level. The activity of caspase 3 in K562 cells was increased by As(2)S(2). As(2)S(2) also induced apoptosis of fresh mononuclear cells derived from chronic myelogenous leukemia (CML) patients.
CONCLUSIONAs(2)S(2) can induce apoptosis of CML cells. The decline of Bcr-Abl may play an important role. The upregulation of bax, increase of the activity of caspase 3, downregulation of c-myc and decrease of JAK2 may also be involved in the mechanism.
Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Caspase 3 ; metabolism ; Fusion Proteins, bcr-abl ; analysis ; Humans ; Janus Kinase 2 ; analysis ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; pathology ; Sulfides ; pharmacology ; bcl-2-Associated X Protein ; analysis
10.Research Advance of BCR-ABL Mutation and the Efficacy of Second and Third Generation TKI in Chronic Myeloid Leukemia--Review.
Journal of Experimental Hematology 2023;31(2):585-588
The treatment of chronic myeloid leukemia (CML) was revolutionized with the advent of the first-generation tyrosine kinase inhibitors (TKIs), but drug resistance developed during treatment, leading to the development of the second-generation (dasatinib, nilotinib, and bosutinib) and third-generation (ponatinib) TKI. Compared with previous treatment regimens, specific TKI can significantly improve the response rate, overall survival rate and prognosis of CML. Only a few patients with BCR-ABL mutation are insensitive to the second-generation TKIs, so it is suggested to select the second-generation TKIs for patients with specific mutations. For patients with other mutations and without mutations, the second-generation TKI should be selected according to the patient's medical history, while the third-generation TKIs should be selected for mutations that are insensitive to the second-generation TKIs, such as T315I mutation that is sensitive to ponatinib. Due to different BCR-ABL mutations in patients with different sensitivity to the second and third-generation TKIs, this paper will review the latest research progress of the efficacy of the second and third-generation TKIs in CML patients with BCR-ABL mutations.
Humans
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Antineoplastic Agents/pharmacology*
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Dasatinib/pharmacology*
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Drug Resistance, Neoplasm/genetics*
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Fusion Proteins, bcr-abl/genetics*
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Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy*
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Mutation
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Protein Kinase Inhibitors/therapeutic use*