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Hiroshi Miwa,
Masato Shikami,
Mineaki Goto,
Shohei Mizuno,
Miyuki Takahashi,
Norikazu Tsunekawa-Imai,
Takamasa Ishikawa,
Motonori Mizutani,
Tomohiro Horio,
Mayuko Gotou, Hidesuke Yamamoto,
Motohiro Wakabayashi,
Masaya Watarai,
Ichiro Hanamura,
Akira Imamura,
Hidetsugu Mihara,
Masakazu Nitta
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ABSTRACT: The shift in energy metabolism from oxidative phosphorylation to glycolysis can serve as a target for the inhibition of cancer growth. Here, we examined the metabolic changes induced by 2-deoxyglucose (2-DG), a glycolysis inhibitor, in leukemia cells by metabolome analysis. NB4 cells mainly utilized glucose as an energy source by glycolysis and oxidative phosphorylation in mitochondria, since metabolites in the glycolytic pathway and in the tricarboxylic acid (TCA) cycle were significantly decreased by 2-DG. In THP-1 cells, metabolites in the TCA cycle were not decreased to the same extent by 2-DG as in NB4 cells, which indicates that THP-1 utilizes energy sources other than glucose. TCA cycle metabolites in THP-1 cells may be derived from acetyl-CoA by fatty acid β-oxidation, which was supported by abundant detection of carnitine and acetylcarnitine in THP-1 cells. 2-DG treatment increased the levels of pentose phosphate pathway (PPP) metabolites and augmented the generation of NADPH by glucose-6-phosphate dehydrogenase. An increase in NADPH and upregulation of glutathione synthetase expression resulted in the increase in the reduced form of glutathione by 2-DG in NB4 cells. We demonstrated that a combination of 2-DG and inhibition of PPP by dehydroepiandrosterone (DHEA) effectively suppressed the growth of NB4 cells. The replenishment of the TCA cycle by fatty acid oxidation by carnitine palmitoyltransferase in THP-1 cells, treated by 2-DG, might be regulated by AMPK, as the combination of 2-DG and inhibition of AMPK by compound C potently suppressed the growth of THP-1 cells. Although 2-DG has been effective in preclinical and clinical studies, this treatment has not been fully explored due to concerns related to potential toxicities such as brain toxicity at high doses. We demonstrated that a combination of 2-DG and DHEA or compound C at a relatively low concentration effectively inhibits the growth of NB4 and THP-1 cells, respectively. These observations may aid in the identification of appropriate combinations of metabolic inhibitors at low concentrations which do not cause toxicities.
Oncology Reports 02/2013; · 1.84 Impact Factor
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Mayuko Gotou,
Ichiro Hanamura,
Hisao Nagoshi,
Motohiro Wakabayashi,
Natsumi Sakamoto,
Norikazu Tsunekawa,
Tomohiro Horio,
Mineaki Goto,
Shohei Mizuno,
Miyuki Takahashi, [......], Hidesuke Yamamoto,
Akihito Hiramatsu,
Masaya Watarai,
Masato Shikami,
Akira Imamura,
Hidetsugu Mihara,
Tomohiko Taki,
Hiroshi Miwa,
Masafumi Taniwaki,
Masakazu Nitta
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ABSTRACT: In this study, we established and analyzed a novel human myeloid leukemia cell line, AMU-AML1, from a patient with acute myeloid leukemia with multilineage dysplasia before the initiation of chemotherapy. AMU-AML1 cells were positive for CD13, CD33, CD117, and HLA-DR by flow cytometry analysis and showed a single chromosomal abnormality, 46, XY, t(12;22)(p13;q11.2), by G-banding and spectral karyotyping. Fluorescent in situ hybridization analysis indicated that the chromosomal breakpoint in band 12p13 was in the sequence from the 5' untranslated region to intron 1 of TEL and that the chromosomal breakpoint in band 22q11 was in the 3' untranslated region of MN1. The chimeric transcript and protein of MN1-TEL could not be detected by reverse-transcriptase polymerase chain reaction or Western blot analysis. However, the MN1 gene was amplified to three copies detected by array comparative genomic hybridization analysis, and the expression levels of the MN1 transcript and protein were high in AMU-AML1 cells when compared with other cell lines with t(12;22)(p13;q11-12). Our data showed that AMU-AML1 cells contain t(12;22)(p13;q11.2) without chimeric fusion of MN1 and TEL. The AMU-AML1 cells gained MN1 copies and had high expression levels of MN1. Thus, the AMU-AML1 cell line is useful for studying the biological consequences of t(12;22)(p13;q11.2) lacking chimeric MN1-TEL.
Genes Chromosomes and Cancer 10/2011; 51(1):42-53. · 3.31 Impact Factor
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Mayuko Gotou,
Ichiro Hanamura,
Hisao Nagoshi,
Motohiro Wakabayashi,
Natsumi Sakamoto,
Norikazu Tsunekawa,
Tomohiro Horio,
Mineaki Goto,
Shohei Mizuno,
Miyuki Takahashi, [......], Hidesuke Yamamoto,
Akihito Hiramatsu,
Masaya Watarai,
Masato Shikami,
Akira Imamura,
Hidetsugu Mihara,
Tomohiko Taki,
Hiroshi Miwa,
Masafumi Taniwaki,
Masakazu Nitta
[show abstract]
[hide abstract]
ABSTRACT: In this study, we established and analyzed a novel human myeloid leukemia cell line, AMU-AML1, from a patient with acute myeloid leukemia with multilineage dysplasia before the initiation of chemotherapy. AMU-AML1 cells were positive for CD13, CD33, CD117, and HLA-DR by flow cytometry analysis and showed a single chromosomal abnormality, 46, XY, t(12;22)(p13;q11.2), by G-banding and spectral karyotyping. Fluorescent in situ hybridization analysis indicated that the chromosomal breakpoint in band 12p13 was in the sequence from the 5′ untranslated region to intron 1 of TEL and that the chromosomal breakpoint in band 22q11 was in the 3′ untranslated region of MN1. The chimeric transcript and protein of MN1-TEL could not be detected by reverse-transcriptase polymerase chain reaction or Western blot analysis. However, the MN1 gene was amplified to three copies detected by array comparative genomic hybridization analysis, and the expression levels of the MN1 transcript and protein were high in AMU-AML1 cells when compared with other cell lines with t(12;22)(p13;q11-12). Our data showed that AMU-AML1 cells contain t(12;22)(p13;q11.2) without chimeric fusion of MN1 and TEL. The AMU-AML1 cells gained MN1 copies and had high expression levels of MN1. Thus, the AMU-AML1 cell line is useful for studying the biological consequences of t(12;22)(p13;q11.2) lacking chimeric MN1-TEL. © 2011 Wiley Periodicals, Inc.
Genes Chromosomes and Cancer 10/2011; 51(1):42 - 53. · 3.31 Impact Factor
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Kazuto Suganuma,
Hiroshi Miwa,
Norikazu Imai,
Masato Shikami,
Mayuko Gotou,
Mineaki Goto,
Shohei Mizuno,
Miyuki Takahashi, Hidesuke Yamamoto,
Akihito Hiramatsu,
Motohiro Wakabayashi,
Masaya Watarai,
Ichiro Hanamura,
Akira Imamura,
Hidetsugu Mihara,
Masakazu Nitta
[show abstract]
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ABSTRACT: For generation of energy, cancer cells utilize glycolysis more vigorously than oxidative phosphorylation in mitochondria (Warburg effect). We examined the energy metabolism of four leukemia cell lines by using glycolysis inhibitor, 2-deoxy-d-glucose (2-DG) and inhibitor of oxidative phosphorylation, oligomycin. NB4 was relatively sensitive to 2-DG (IC(50): 5.75 mM), consumed more glucose and produced more lactate (waste product of glycolysis) than the three other cell lines. Consequently, NB4 was considered as a "glycolytic" leukemia cell line. Dependency on glycolysis in NB4 was confirmed by the fact that glucose (+) FCS (-) medium showed more growth and survival than glucose (-) FCS (+) medium. Alternatively, THP-1, most resistant to 2-DG (IC(50): 16.14 mM), was most sensitive to oligomycin. Thus, THP-1 was recognized to be dependent on oxidative phosphorylation. In THP-1, glucose (-) FCS (+) medium showed more growth and survival than glucose (+) FCS (-) medium. The dependency of THP-1 on FCS was explained, at least partly, by fatty acid oxidation because inhibitor of fatty acid β-oxidation, etomoxir, augmented the growth suppression of THP-1 by 2-DG. We also examined the mechanisms by which THP-1 was resistant to, and NB4 was sensitive to 2-DG treatment. In THP-1, AMP kinase (AMPK), which is activated when ATP becomes limiting, was rapidly phosphorylated by 2-DG, and expression of Bcl-2 was augmented, which might result in resistance to 2-DG. On the other hand, AMPK phosphorylation and augmentation of Bcl-2 expression by 2-DG were not observed in NB4, which is 2-DG sensitive. These results will facilitate the future leukemia therapy targeting metabolic pathways.
Leukemia & lymphoma 11/2010; 51(11):2112-9. · 2.40 Impact Factor
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ABSTRACT: Thrombosis in myeloproliferative thrombocytosis occurs usually in the microvessels and medium-sized arteries and veins and only rarely in the aorta. Aortic thrombosis is usually treated with thrombectomy. Reported here is a rare case that was treated pharmacologically.
A 60-year-old Japanese woman presented with numbness of both lower extremities. Her platelet count was 1787 x 103/mul. Through bone marrow examination, we diagnosed her condition as myelodysplastic and/or myeloproliferative disorder-unclassifiable. Abdominal ultrasonography and computed tomographic scan revealed aortic thrombosis. Her platelet count was controlled with hydroxyurea and ranimustine. Aspirin and ticlopidine improved the numbness in both lower limbs on the second day. Aortic thrombosis was not observed in a computed tomographic scan on the seventh day.
For aortic thrombosis, surgical management is usually adopted, but pharmacological management is also an option because of its immediate curative effects.
Journal of Medical Case Reports 01/2010; 4:219.
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ABSTRACT: Endothelial progenitor cells (EPCs) have been isolated from peripheral blood, bone marrow, and umbilical cord blood (CB) and determined to be in heterogeneous populations; however, specific variations in their characteristics remain to be clarified. In this study, we observed that mononuclear cells (MNCs) of CB change in morphology to differentiate into mature endothelial cells (EC) after 6 weeks of culture. In early days of culture along with the differentiation, two distinct populations of EPCs were detected, defined by two-dimensional dot plots (forward scatter vs side scatter) with flow cytometry, namely, relatively small cells (S-EPCs) and relatively large cells (L-EPCs). S-EPCs were found to express CD34 but not CD14, while the converse was the case for L-EPCs. When CD34(+)/CD14(-) cells and CD34(-)/CD14(+) cells were isolated from original MNCs of CB and cultured independently, S-EPCs and L-EPCs were derived from CD34(+)/CD14(-) and from CD34(-)/CD14(+) cells, respectively. Furthermore, when the two EPCs at day 7 were separated by cell sorter and recultured, there was no crossover in terms of CD34 and CD14 expression. While expression of VE-cadherin and vascular endothelial growth factor receptor-2 (VEGFR-2) on L-EPCs was significantly greater than on S-EPCs, levels of CD31 were lower. In addition, L-EPCs exhibited greater proliferative ability on stimulation with VEGF. Although these two EPCs expressed different phenotypes, including growth factor receptors, and had different proliferative ability, they both eventually differentiated into mature ECs after more than 3 weeks of culture.
Annals of Hematology 03/2008; 87(2):87-95. · 2.62 Impact Factor
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Toshiko Ikai,
Hiroshi Miwa,
Masato Shikami,
Akihito Hiramatsu,
Emi Tajima, Hidesuke Yamamoto,
Norikazu Imai,
Akiko Hattori,
Kazuhiro Nishii,
Kazuhisa Miura,
Atsushi Satoh,
Masato Itoh,
Akira Imamura,
Hidetsugu Mihara,
Yoshiro Katoh,
Masakazu Nitta
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ABSTRACT: Vascular endothelial growth factor (VEGF) and its associated molecule, placenta growth factor (PlGF) are now known to support normal hematopoiesis, and leukemia cell growth. In this study, expression of VEGF and PlGF in acute lymphoblastic leukemia (ALL) cells was examined by real time reverse transcription-polymerase chain reaction in 20 patient samples. Expression of PlGF was more intense in Philadelphia chromosome positive (Ph(+)) ALL than in Ph(-) ALL cases. On the other hand, expression level of VEGF was not different between Ph(+) and Ph(-) cases. Then, PlGF was added to the two ALL cell lines, CRL1929 (Ph(+)), and Nalm6 (Ph(-)). The PlGF stimulated the growth of CRL1929 in time- and dose-dependent manners, although the growth of Nalm6 was not affected by PlGF. The growth stimulation of CRL1929 by PlGF was confirmed by the increase of S phase cells. And the growth promoting effect of PlGF on CRL1929 was cancelled by simultaneous addition of VEGFR1/Fc (which binds to PlGF and abrogates its function), but was not cancelled by VEGFR2/Fc (which does not bind to PlGF). Then, addition of VEGFR1/Fc to the simple culture of CRL1929 demonstrated growth inhibitory effect. These observations demonstrated that PlGF stimulates the growth of Ph(+) ALL cells by both autocrine and paracrine pathways. Finally, PlGF-VEGFR1 loop might be a therapeutic target to improve the prognosis of Ph(+) ALL.
European Journal Of Haematology 11/2005; 75(4):273-9. · 2.61 Impact Factor
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ABSTRACT: We report an unusual case of angioimmunoblastic T cell lymphoma arising in the setting of 5 years of immunosuppressive treatment for progressive systemic sclerosis. The lymph node lesion was accompanied by large blastic B cells with an association of Epstein-Barr virus. Southern blot study demonstrated the clonal rearrangement of T cell receptor beta-chain gene, but not of immunoglobulin heavy chain gene. Phenotypical examination of the lymph node also revealed the predominance of CD4+ T cells in addition to the proliferation of follicular dendritic cells, but no light chain restriction in large B cell components. In the clinical and laboratory aspects, neutrophilia (15.8 x 10(9)/l) and plasmacytosis (40%) in bone marrow were noted, which were considered to be closely related to elevated serum granulocyte colony-stimulating factor, interleukin (IL)-4 and IL-6. Based on the combined data described here, our preferred diagnosis was angioimmunoblastic T cell lymphoma with Epstein-Barr virus-associated B cell lymphoproliferative disorder, the pathogenesis of which was suggested to be closely associated with immunosuppressive treatment for progressive systemic sclerosis.
Acta Haematologica 02/2005; 114(2):108-12. · 1.35 Impact Factor
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Motohiro Wakabayashi,
Hiroshi Miwa,
Masato Shikami,
Akihito Hiramatsu,
Toshiko Ikai,
Emi Tajima, Hidesuke Yamamoto,
Kazuhisa Miura,
Atsushi Satoh,
Masato Itoh,
Akira Imamura,
Hidetsugu Mihara,
Yoshiro Katoh,
Masakazu Nitta
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ABSTRACT: Hematopoietic cells and endothelial cells are mutually correlated in their development and growth. Various angiogenic factors, such as vascular endothelial growth factor (VEGF) and angiopoietins (Angs), are thought to be associated with leukemia cell growth. In this study, we examined if the Angs-Tie2 autocrine pathway works in primary AML cells or not by using soluble Tie2-Fc, which inhibits Angs from binding to Tie2 receptor. After 48 h of culture with Tie2-Fc, nine AML cells from 19 examined samples were not influenced by Tie2-Fc (group A), while AML cells from remaining 10 patients demonstrated remarkable reduction of cell number by Tie2-Fc treatment (group B). Tie2 receptor, upon binding to Angs, are known to activate phosphatidyl-inositol 3 kinase (PI3 kinase). Then, we examined the effect of LY294002, a potent PI3 kinase inhibitor, on primary AML cells. Cell number reduction effect by the treatment of LY294002 was much more prominent in cells of group B than of group A. In addition, extent of cell number reduction by Tie2-Fc and LY294002 was quite well correlated. These observations demonstrated that cells from a part of AML were dependent on autocrine Angs-Tie2 pathway. This notion was further supported by the study of two AML cell lines, KG-1 and HL-60: the growth of KG-1 was suppressed by Tie2-Fc, and also by anti-Tie2 antibody, which inhibits receptor-ligand interaction, while that of HL-60 was not suppressed by Tie2-Fc or anti-Tie2 antibody. Our results will help to explore the angiogenesis-oriented or endothelial cell-mediated therapy for leukemia.
The Hematology Journal 02/2004; 5(4):353-60. · 1.86 Impact Factor
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Atsushi Satoh,
Toshiko Ikai,
Hiroshi Miwa,
Norikazu Imai,
Akihito Hiramatsu,
Emi Tajima, Hidesuke Yamamoto,
Motohiro Wakabayashi,
Kazuhisa Miura,
Masato Ito,
Masato Shikami,
Akira Imamura,
Hidetsugu Mihara,
Yoshiro Kato,
Masakazu Nitta
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ABSTRACT: We report a case of acute promyelocytic leukemia (APL) with drug-induced hypersensitivity syndrome associated with Epstein-Barr virus (EBV) infection. A 33-year-old woman was admitted because of APL. After complete remission was obtained with the use of all-trans retinoic acid (ATRA), intensive chemotherapy was administered. She developed high grade fever and severe systemic erythematous eruptions followed by cervical lymphoadenopathy, hepatosplenomegaly, hepatitis and hypotension in a state of myelosuppression during consolidation chemotherapy. Systemic corticosteroids alleviated the symptoms. Since an anti-EB VCA IgM antibody titer was continuously positive, persistent infection of EBV was suspected. In this case, EBV infection may have contributed to the development of drug-induced hypersensitivity syndrome.
Internal Medicine 02/2004; 43(1):74-8. · 0.94 Impact Factor
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ABSTRACT: We describe a patient who presented with aplastic anaemia associated with the Philadelphia (Ph1) chromosome during immunosuppressive therapy and who subsequently developed myelodysplastic syndrome (MDS) with monosomy 7. Initially the patient had hypocellular fatty marrow without leukaemic blasts or dysplastic features. Chromosome analysis showed 46, XY, t(9;22)(q34;q11) during immunosuppressive therapy, but no leukaemic transformation was detected. The patient showed gradual haematologic improvement and became transfusion independent. Thereafter, bone marrow dysplasia with monosomy 7 progressed following transfusion independence. These findings indicate that multiple cytogenetic evolutions occur in aplastic anaemia during immunosuppressive therapy, and that Ph1 chromosome may play a role in bone marrow suppression rather than development of leukaemia.
European Journal Of Haematology 09/2003; 71(2):130-2. · 2.61 Impact Factor
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ABSTRACT: We describe a patient who presented with aplastic anaemia associated with the Philadelphia (Ph1) chromosome during immunosuppressive therapy and who subsequently developed myelodysplastic syndrome (MDS) with monosomy 7. Initially the patient had hypocellular fatty marrow without leukaemic blasts or dysplastic features. Chromosome analysis showed 46, XY, t(9;22)(q34;q11) during immunosuppressive therapy, but no leukaemic transformation was detected. The patient showed gradual haematologic improvement and became transfusion independent. Thereafter, bone marrow dysplasia with monosomy 7 progressed following transfusion independence. These findings indicate that multiple cytogenetic evolutions occur in aplastic anaemia during immunosuppressive therapy, and that Ph1 chromosome may play a role in bone marrow suppression rather than development of leukaemia.
European Journal Of Haematology 07/2003; 71(2):130 - 132. · 2.61 Impact Factor
