Synergistically Cooperation of Bortezomib and Arsenic Trioxide on Chronic Myelogenous Leukemia

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4233-4233
Author(s):  
Hua Yan ◽  
Dao Li ◽  
Li Chen ◽  
Ying-Li Wu ◽  
Jun-Ming Li ◽  
...  
Keyword(s):  
Cell Growth ◽  
Arsenic Trioxide ◽  
Chronic Myelogenous Leukemia ◽  
Combined Treatment ◽  
Flow Cytometric Analysis ◽  
Chemotherapeutic Agents ◽  
Myelogenous Leukemia ◽  
Imatinib Treatment ◽  
Single Treatment ◽  
Flow Cytometric

Abstract Chronic Myelogenous Leukemia (CML) is one of myeloproliferative disorders characterized by the chromosome translocation (9; 22), which causes the generation of the Bcr-Abl fusion protein. Latter activates a number of signal transducers and transcription factors, thus leading to multi-resistance to chemotherapeutic agents. Introduction of imatinib mesylate (Gleevec) significantly increased CML patients’ response but still some of patients got disease progression. Therefore, it still remains a high priority to develop new approaches to treat Bcr-Abl+ leukemia. Aim: We aim to investigate the synergy of bortezomib and arsenic trioxide on inducing the apoptosis on Bcr-Abl+ K562-s as well as K562-r cells, which was sensitive or resistant to imatinib treatment respectively. Material and methods: Bcr-Abl+ K562-s and K562-r cells were chosen to evaluate the drugs’ effect. We observed the inhibition of cell growth and cell viability after bortezomib and/or arsenic treatment. Flow cytometric evaluation and western blot analysis were performed to detect the development of apoptosis, the changeable expression of Bcr-Abl protein and the apoptosis-related proteins. Results: K562-s and K562-r cells had different response to imatinib treatment. As expected, 0.25μM of imatinib can inhibit half of K562-s cell growth whereas 10–15 more times of concentration of imatinib were necessary to reach the same inhibitory effect in K562-r cells. Combined bortezomib at 12nM with 1μM of ATO, the concentration inducing 50% of K562-s growth arrest, synergistically induced apoptosis in K562-s cells after 48hrs of cotreatment, confirmed by notable elevation of Annexin V+ cells through flow cytometric analysis, when compared to those in the control or single treatment (p<0.01), and immunoblotting detection of caspase-3 cleavage, degradation of poly-adenosine diphosphate-ribose polymerase (PARP), and decreased expression of Bcr-Abl protein. More surprisingly, 24nM concentration of bortezomib and 2μM of ATO, 2 times of concentration needed for 50% of K562-s cell growth inhibition respectively, inhibited 50% of K562-r cells growth, far less than the increased proportion needed during imatinib treatment. And more of important, although single treatment of bortezomib or ATO had no or little effect on inducing apoptosis, combined treatment significantly induced the apoptosis in K562-r cells (p<0.05), associated with activation of apoptosis-relevant proteins and obvious degradation of Bcr-Abl protein. Conclusion: Taken together, these findings indicate that bortezomib combined with ATO could be developed into a novel therapeutic strategy for CML. More study will be needed to further explore its potent mechanism involved during cotreatment.

Glutathione-Depletion Potently Enhances the Cytotoxic Activity of Arsenic Trioxide in Imatinib Sensitive and Resistant Bcr-Abl Positive Cell Lines.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2098-2098
Author(s):  
Heiko Koenig ◽  
Erika M. Moseson ◽  
Christian Lorentz ◽  
Peter Paschka ◽  
Junia V. Melo ◽  
...  
Keyword(s):  
Arsenic Trioxide ◽  
Cell Lines ◽  
Flow Cytometric Analysis ◽  
Myelogenous Leukemia ◽  
Glutathione Depletion ◽  
Similar Data ◽  
Antileukemic Activity ◽  
Treatment Needs ◽  
Flow Cytometric ◽  
Proliferation Assays

Abstract Arsenic trioxide (ATO, Trisenox™) is currently being investigated as a potential drug to optimize imatinib based treatment of chronic myelogenous leukemia (CML). ATO has antileukemic activity against imatinib sensitive and resistant CML cell lines and primary cells derived from patients with CML. However, preliminary clinical data reveal only moderate activity of ATO in CML (imatinib sensitive or resistant). ATO-activity in acute myelogenous leukemia or multiple myeloma cells is inversely related to the intracellular Glutathione- (GSH-) content with cells bearing low GSH-levels being the most sensitive. To investigate whether this relationship plays a role in CML cell lines sensitive or resistant to the Bcr-Abl inhibitor imatinib, we determined the antiproliferative activity of ATO with regard to the cellular GSH-levels. Furthermore, we examined the ability of GSH-depletion as a therapeutic tool to sensitize CML cells to ATO. MTS-proliferation assays were performed to determine the concentration of ATO needed to induce 50% cellular growth inhibition (IC50). Cell lines used were the imatinib sensitive CML blast crisis lines AR230-s, KCL22-s, LAMA-s as well as their imatinib resistant derivatives AR230-r1, KCL22-r1, LAMA-r1. Known ATO-sensitive AML cell lines (NB4, HL60) with low GSH-content were also included in this study. Intracellular GSH-levels were measured biochemically using a commercially available kit. Protein content was analyzed using the Bradford method. ATO toxicity was analyzed using trypan blue exclusion and flow cytometric analysis of Annexin/PI-stained cells. MTS-proliferation assays indicate a cell type dependent activity of ATO with IC50-values ranging from 0.37±0.03 μM (NB4) up to 6.9±1.43 μM (AR230-r1). The most sensitive NB4 cells express low GSH-levels (8.74±2.9 nmol/mg), whereas highly resistant AR230-r1 cells reveal a threefold increased GSH-content (26.7±8nmol/mg). Imatinib resistance in AR230-r1 and LAMA-r1 cells is not associated with significant GSH-content modulation when compared to the imatinib naïve counterparts (23.6±5.4nmol/mg [AR230-s] vs 26.7±8nmol/mg [AR230-r1]; 8.3±1.8nmol/mg [LAMA-s] vs 6.9±0.2 [LAMA-r1]). Treatment of AR230-s cells with 100 μM of the GSH-depleting agent L-Buthionine-Sulfoximine (BSO) for 12 h leads to significant downregulation of cellular GSH (23.6±5.4 nmol/mg [control] vs 4,25±0,53 nmol/mg [100 μM BSO]). Treatment with BSO alone does not affect cellular viability nor induces apoptosis. Subsequent cotreatment of AR230-s cells with ATO (1 μM) and BSO (100 μM) for 24 h reduces viability to 31.6 % compared to untreated cells. In contrast, treatment with 1 μM ATO alone does not affect viability. Flow cytometric analysis of apoptosis reflects viability data with a 5.5 fold increase of apoptotic cells in the combined treated fraction. Similar data were generated using the KCL22-s cell line. Experiments using primary patient cells are currently in progress. Our data indicate that high intracellular GSH-content confers relative resistance to ATO in Bcr-Abl positive cell lines regardless whether they are imatinib sensitive or resistant. A promising tool to increase the antileukemic activity of ATO is the application of GSH-depleting agents. Therefore, GSH-dependent response to ATO treatment needs further investigation in individual CML patients.

Humoral-Dependent Hemopoiesis and Flow Cytometric Analysis of Chronic Myelogenous Leukemia in Erythroblastic Transformation

1987 ◽  
Vol 77 (2) ◽  
pp. 120-123 ◽  
Author(s):  
J.D. Lutton ◽  
J.W. Chiao ◽  
J.L. Ascensao ◽  
M. Atamer ◽  
Z. Arlin ◽  
...  
Keyword(s):  
Chronic Myelogenous Leukemia ◽  
Flow Cytometric Analysis ◽  
Myelogenous Leukemia ◽  
Flow Cytometric

scholarly journals Autophagic degradation of the BCR-ABL oncoprotein and generation of antileukemic responses by arsenic trioxide

Blood ◽  
2012 ◽  
Vol 120 (17) ◽  
pp. 3555-3562 ◽  
Author(s):  
Dennis J. Goussetis ◽  
Elias Gounaris ◽  
Edward J. Wu ◽  
Eliza Vakana ◽  
Bhumika Sharma ◽  
...  
Keyword(s):  
Arsenic Trioxide ◽  
Chronic Myelogenous Leukemia ◽  
Cathepsin B ◽  
Myelogenous Leukemia ◽  
Molecular Targeting ◽  
Therapeutic Approaches ◽  
Subsequent Degradation ◽  
Autophagic Degradation ◽  
Partial Reversal

Abstract We provide evidence that arsenic trioxide (As2O3) targets the BCR-ABL oncoprotein via a novel mechanism involving p62/SQSTM1-mediated localization of the oncoprotein to the autolysosomes and subsequent degradation mediated by the protease cathepsin B. Our studies demonstrate that inhibitors of autophagy or cathepsin B activity and/or molecular targeting of p62/SQSTM1, Atg7, or cathepsin B result in partial reversal of the suppressive effects of AS2O3 on BCR-ABL expressing leukemic progenitors, including primitive leukemic precursors from chronic myelogenous leukemia (CML) patients. Altogether, these findings indicate that autophagic degradation of BCR-ABL is critical for the induction of the antileukemic effects of As2O3 and raise the potential for future therapeutic approaches to target BCR-ABL expressing cells by modulating elements of the autophagic machinery to promote BCR-ABL degradation.

After chronic myelogenous leukemia: tyrosine kinase inhibitors in other hematologic malignancies

Blood ◽  
2005 ◽  
Vol 105 (1) ◽  
pp. 22-30 ◽  
Author(s):  
Martha Wadleigh ◽  
Daniel J. DeAngelo ◽  
James D. Griffin ◽  
Richard M. Stone
Keyword(s):  
Cell Growth ◽  
Small Molecules ◽  
Chronic Myelogenous Leukemia ◽  
Tyrosine Kinases ◽  
Kinase Inhibitors ◽  
Hematologic Malignancies ◽  
Myelogenous Leukemia ◽  
Pathologic Feature ◽  
Growth And Survival ◽  
Tyrosine Residues

AbstractTyrosine kinases phosphorylate proteins on tyrosine residues, producing a biologic signal that influences many aspects of cellular function including cell growth, proliferation, differentiation, and death. Constitutive or unregulated activity through mutation or overexpression of these enzymes is a common pathologic feature in many acute and chronic leukemias. Inhibition of tyrosine kinases represents a strategy to disrupt signaling pathways that promote neoplastic growth and survival in hematologic malignancies and likely in other neoplasias as well. This review focuses on tyrosine kinases that have been implicated in the pathogenesis of hematologic diseases other than chronic myelogenous leukemia and discusses the evidence for the use of small molecules to target these kinases.

Immunophenotypic Study of Basophils by Multiparameter Flow Cytometry

2008 ◽  
Vol 132 (5) ◽  
pp. 813-819
Author(s):  
Xiaohong Han ◽  
Jeffrey L. Jorgensen ◽  
Archana Brahmandam ◽  
Ellen Schlette ◽  
Yang O. Huh ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Chronic Myelogenous Leukemia ◽  
Peripheral Blood ◽  
Myelogenous Leukemia ◽  
Multiparameter Flow Cytometry ◽  
Color Flow ◽  
Aberrant Expression ◽  
Flow Cytometric ◽  
Hla Dr

Abstract Context.—The immunophenotypic profile of basophils is not yet fully established, and the immunophenotypic changes in chronic myelogenous leukemia are not fully characterized. Objective.—To establish a comprehensive immunophenotypic spectrum of normal basophils and to assess the range of immunophenotypic aberrations of basophils in chronic myelogenous leukemia. Design.—Using 4-color flow cytometry, we compared the immunophenotypic profile of basophils in peripheral blood or bone marrow samples from 20 patients with no evidence of neoplasia to basophils from 15 patients with chronic myelogenous leukemia. Results.—Basophils in control cases were all positive for CD9, CD13, CD22, CD25 (dim), CD33, CD36, CD38 (bright), CD45 (dimmer than lymphocytes and brighter than myeloblasts), and CD123 (bright), and were negative for CD19, CD34, CD64, CD117, and HLA-DR. Basophils in all chronic myelogenous leukemia patients possessed 1 to 5 immunophenotypic aberrancies. The most common aberrancies were underexpression of CD38, followed by aberrant expression of CD64 and underexpression of CD123. CD34 and CD117 were present in cases with basophilic precursors. Myeloblasts showed a distinct immunophenotypic profile, as they typically expressed CD34 and CD117, showed dimmer expression (compared with basophils) of CD38, CD45, and CD123, and lacked expression of CD22. Conclusions.—Flow cytometric immunophenotyping can identify immunophenotypic aberrations of basophils in chronic myelogenous leukemia, and discriminate basophils from myeloblasts.

scholarly journals BCR-ABL maintains resistance of chronic myelogenous leukemia cells to apoptotic cell death [published erratum appears in Blood 1994 Jun 15;83(12):3835]

Blood ◽  
1994 ◽  
Vol 83 (5) ◽  
pp. 1179-1187 ◽  
Author(s):  
A McGahon ◽  
R Bissonnette ◽  
M Schmitt ◽  
KM Cotter ◽  
DR Green ◽  
...  
Keyword(s):  
Cell Death ◽  
Chronic Myelogenous Leukemia ◽  
Apoptotic Cell ◽  
Chimeric Protein ◽  
Chemotherapeutic Agents ◽  
Myelogenous Leukemia ◽  
Temperature Sensitive ◽  
Permissive Temperature ◽  
Expression Of Genes ◽  
Induction Of Apoptosis

Abstract Apoptosis is the major form of cell death associated with the action of chemotherapeutic agents on tumor cells, and therefore the expression of genes that interfere with apoptosis can have important consequences for the efficacy of therapeutic approaches. Here we show that K562, a chronic myelogenous leukemia (CML) cell line expressing the BCR-ABL fusion protein, are resistant to the induction of apoptosis by a number of agents and conditions. Antisense oligodeoxynucleotides corresponding to the translation start of bcr downregulate bcr-abl protein in these cells and render them susceptible to induction of apoptosis by chemotherapeutic agents or serum deprivation. Expression of a temperature sensitive v-Abl protein reverses the effects of the antisense oligonucleotides, such that the cells remain resistant to apoptosis at the permissive temperature. These data indicate that bcr- abl acts as an anti-apoptosis gene in CML cells and suggests that the effect is dependent on the abl kinase activity in this chimeric protein. Inhibition of bcr-abl to render CML cells susceptible to apoptosis can be combined with therapeutic drugs and/or treatment capable of inducing apoptosis to provide an effective strategy for elimination of these cells.

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