[Show abstract][Hide abstract] ABSTRACT: BACKGROUND: The ABO blood group is important in blood transfusion. Recently, an erythroid cell-specific regulatory element has been identified in the first intron of ABO using luciferase reporter assays with K562 cells. The erythroid cell-specific regulatory activity of the element was dependent upon GATA-1 binding. In addition, partial deletion of Intron 1 including the element was observed in genomic DNAs obtained from 111 Bm and ABm individuals, except for one, whereas the deletion was never found among 1005 individuals with the common phenotypes. STUDY DESIGN AND METHODS: In this study, further investigation was performed to reveal the underlying mechanism responsible for reduction of B antigen expression in the exceptional Bm individual. Peptide nucleic acid-clamping polymerase chain reaction was carried out to amplify the B-related allele, followed by sequence determination. Electrophoretic mobility assays and promoter assays were performed to examine whether a nucleotide substitution reduced the binding of a transcription factor and induced loss of function of the element. RESULTS: Sequence determination revealed one point mutation of the GATA motif in the element. The electrophoretic mobility shift assays showed that the mutation abolished the binding of GATA transcription factors, and the promoter assays demonstrated complete loss of enhancer activity of the element. CONCLUSION: These observations suggest that the mutation in the GATA motif of the erythroid-specific regulatory element may diminish the binding of GATA transcription factors and down regulate transcriptional activity of the element on the B allele, leading to reduction of B antigen expression in erythroid lineage cells of the Bm individual.
[Show abstract][Hide abstract] ABSTRACT: We report a case of adult T-cell leukemia/lymphoma (ATL) with rapidly progressive pulmonary areas of ground-glass attenuation (GGA) and nodules resulting from acute transformation of chronic ATL. A 48-year-old Japanese man was admitted for progressive dyspnea. Chest computed tomography showed rapidly progressive bilateral pulmonary areas of GGA and nodules. Flow cytometry of bronchoalveolar lavage fluid and immunohistochemical examination of lung biopsy specimens revealed invasion of ATL cells. Systemic chemotherapy improved the pulmonary findings. Rapidly expanding areas of GGA and nodules are a rare manifestation of pulmonary invasion of ATL that clinicians should nevertheless keep in mind.
[Show abstract][Hide abstract] ABSTRACT: The ABO blood group is of great importance in blood transfusion and organ transplantation. However, the mechanisms regulating human ABO gene expression remain obscure. On the basis of DNase I-hypersensitive sites in and upstream of ABO in K562 cells, in the present study, we prepared reporter plasmid constructs including these sites. Subsequent luciferase assays indicated a novel positive regulatory element in intron 1. This element was shown to enhance ABO promoter activity in an erythroid cell-specific manner. Electrophoretic mobility-shift assays demonstrated that it bound to the tissue-restricted transcription factor GATA-1. Mutation of the GATA motifs to abrogate binding of this factor reduced the regulatory activity of the element. Therefore, GATA-1 appears to be involved in the cell-specific activity of the element. Furthermore, we found that a partial deletion in intron 1 involving the element was associated with B(m) phenotypes. Therefore, it is plausible that deletion of the erythroid cell-specific regulatory element could down-regulate transcription in the B(m) allele, leading to reduction of B-antigen expression in cells of erythroid lineage, but not in mucus-secreting cells. These results support the contention that the enhancer-like element in intron 1 of ABO has a significant function in erythroid cells.
[Show abstract][Hide abstract] ABSTRACT: We report a 73-year-old Japanese man with early onset pure red cell aplasia (PRCA) caused by subcutaneous administration of recombinant epoetin-β. Two months after the start of epoetin therapy, he developed PRCA. Anti-erythropoietin (EPO) antibody, detected in the patient's serum by enzyme immunoassay and radioimmunoprecipitation method, inhibited EPO-dependent growth of AS-E2 cells in vitro. Treatment with prednisone (1 mg/kg) significantly reduced antibody levels 3 months later. It is important to have an awareness of antibody-mediated PRCA. Our case shows that subcutaneous epoetin administration produces this complication in the early period of therapy.
[Rinshō ketsueki] The Japanese journal of clinical hematology 01/2012; 53(1):110-2.
[Show abstract][Hide abstract] ABSTRACT: Human T-cell leukemia is an aggressive malignancy of T lymphocytes. T-cell leukemia has a very poor prognosis, even with intensive chemotherapy, indicating the need for development of new drugs to treat the disease. Triterpenoid cucurbitacins have been shown to have antitumor activity, but the mechanism of this activity is not fully understood.
The effects of cucurbitacin D on the proliferation and apoptotic induction of T-cell leukemia cells using the Cell viability assay and Annexin V staining were evaluated. To investigate the mechanisms of apoptosis, antiapoptotic protein, NF-κB, and the proteasome activity of leukemia cells treated with cucurbitacin D were evaluated by Western blotting both in vitro and in vivo.
In this study, cucurbitacin D was found to inhibit proliferation and to induce apoptosis of T-cell leukemia cells. Constitutively activated NF-κB was inhibited by cucurbitacin D in the nucleus, which resulted in accumulation of NF-κB in the cytoplasm, leading to down-regulation of the expression of antiapoptotic proteins Bcl-xL and Bcl-2. Furthermore, cucurbitacin D induced the accumulation of inhibitor of NF-κB (IκB)α by inhibition of proteasome activity. Low doses of cucurbitacin D synergistically potentiated the antiproliferative effects of the histone deacetylase inhibitor VPA. Finally, the proapoptotic and proteasome inhibitory activities of cucurbitacin D also were demonstrated using SCID mice in an in vivo study.
Cucurbitacin D induced apoptosis through suppression of proteasome activity both in vitro and in vivo, making cucurbitacin D a promising candidate for clinical applications in the treatment of T-cell leukemia.
Cancer 06/2011; 117(12):2735-46. · 5.20 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The C/EBP family of proteins represents an important group of bZIP transcription factors that are key to the regulation of essential functions such as cell cycle, hematopoiesis, skeletal development, and host immune responses. They are also intimately associated with tumorigenesis and viral disease. These proteins are regulated at multiple levels, including gene induction, alternative translational initiation, post-translational modification, and protein-protein interaction. This review attempts to integrate recent reports with more than 20 years of previous effort focused on this fascinating collection of regulators.
[Show abstract][Hide abstract] ABSTRACT: The efficacy of pirarubicin (THP)-COP was previously compared with cyclophophamide + doxorubicin + vincristine + prednisolone (CHOP) in elderly patients with lymphoma. The subset analysis showed that T-cell lymphoma had a significantly better response with THP-COP, whereas no such difference was observed in B-cell lymphoma. The aim of this study is to confirm the efficacy of THP-COP in the treatment of T-cell lymphoma.
We underwent a multicenter phase II study of THP-COP as a first-line treatment for T-cell lymphoma. The overall response rate, survival period, and toxicity were analyzed.
Fifty-three patients were enrolled in this study. Seventeen patients had peripheral T-cell lymphoma (PTCL), including nine of PTCL not otherwise specified (PTCL-NOS) and eight of angioimmunoblastic T-cell lymphoma (AITL). Thirty-six patients had adult T-cell leukemia/lymphoma (ATLL), including 20 of acute type and 16 of lymphoma type. A treatment response was obtained in 35 (66%) patients, including 17 (32%) complete responses. Median overall survival (OS) and progression-free survival (PFS) times were 14.3 months and 5.2 months, respectively. Patients with ATLL showed a tendency to obtain low response rate (61% vs. 77%, P = 0.27) and had a significantly inferior OS (13.3 vs. 28.6 months, P = 0.04) and PFS (4.6 vs. 8.1 months, P = 0.01) in comparison with PTCL. Grade 3 to 4 neutropenia, anemia, and thrombocytopenia occurred in 72%, 34%, and 58% of the patients, respectively. Febrile neutropenia was observed in 51% and grade 3 non-hematological toxicities in 2-9% of the patients.
The efficacy of THP-COP is equivalent to that of CHOP for the first-line therapy in T-cell lymphoma.
European Journal Of Haematology 05/2010; 84(5):391-7. · 2.55 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Whether galectin-9 plays a role in inflammatory responses remains elusive. The present study was designed to determine the role of intracellular galectin-9 in activation of inflammatory cytokine genes in human monocytes. Galectin-9 expression vector pBKCMV3-G9 was transiently co-transfected into THP-1 monocytic cells along with luciferase reporters carrying gene promoters of IL-1alpha (IL1A), IL-1beta (IL1B) and IFNgamma. Transient transfection studies showed that galectin-9 over-expression activated all three gene promoters, suggesting that intracellular galectin-9 induces inflammatory cytokine genes in monocytes. Galectin-9 over-expression also activated NF-IL6 (C/EBP beta) and AP-1, but not NF-kappaB. In contrast, extracellular galectin-9 is not involved in regulation of inflammatory cytokines. Immunoprecipitation/Western blotting, using anti-galectin-9 Ab and anti-NF-IL6 Ab, showed physical association of intracellular galectin-9 with NF-IL6. RT-PCR confirmed that galectin-9 over-expression increased IL-1alpha and IL-1beta mRNA levels in THP-1 cells. The interaction of galectin-9 with NF-IL6 was enhanced following LPS treatment in THP-1 cells. Intracellular galectin-9 synergized with LPS to activate NF-IL6. Nuclear translocation of galectin-9 was also observed in THP-1 cells treated with LPS. Our results indicate that galectin-9 is a LPS-responsive factor, and further demonstrate that intracellular galectin-9 transactivates inflammatory cytokine genes in monocytes through direct physical interaction with NF-IL6.
Genes to Cells 05/2009; 14(4):511-21. · 2.73 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The oncogenic Wip1 phosphatase (PPM1D) is induced upon DNA damage in a p53-dependent manner and is required for inactivation or suppression of DNA damage-induced cell cycle checkpoint arrest and of apoptosis by dephosphorylating and inactivating phosphorylated Chk2, Chk1, and ATM kinases. It has been reported that arsenic trioxide (ATO), a potent cancer chemotherapeutic agent, in particular for acute promyelocytic leukemia, activates the Chk2/p53 pathway, leading to apoptosis. ATO is also known to activate the p38 MAPK/p53 pathway. Here we show that phosphatase activities of purified Wip1 toward phosphorylated Chk2 and p38 in vitro are inhibited by ATO in a dose-dependent manner. Furthermore, DNA damage-induced phosphorylation of Chk2 and p38 in cultured cells is suppressed by ectopic expression of Wip1, and this Wip1-mediated suppression can be restored by the presence of ATO. We also show that treatment of acute promyelocytic leukemia cells with ATO resulted in induction of phosphorylation and activation of Chk2 and p38 MAPK, which are required for ATO-induced apoptosis. Importantly, this ATO-induced activation of Chk2/p53 and p38 MAPK/p53 apoptotic pathways can be enhanced by siRNA-mediated suppression of Wip1 expression, further indicating that ATO inhibits Wip1 phosphatase in vivo. These results exemplify that Wip1 is a direct molecular target of ATO.
Journal of Biological Chemistry 08/2008; 283(27):18969-79. · 4.65 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Cancer chemotherapy regimens which had been used at a ward and outpatient chemotherapy in various departments were collected and made available to everybody in October, 2003. However, it was difficult to manage cancer chemotherapy regimens in real time, from the viewpoint of risk management. Then, the Cancer Chemotherapy Center took the leading part and established a chemotherapy exploratory committee, which consists of 4 doctors, 2 nurses and 2 pharmacists, in June, 2006. The department of pharmacy could control all cancer chemotherapy regimens by this system, and lead the proper use of increasing anticancer agents. Inquiries on prescription by the pharmacist contributed to proper medical treatment. The role of the cancer chemotherapy exploratory committee and its outcome are described for the purpose of the prevention of medical accidents in this paper.
[Show abstract][Hide abstract] ABSTRACT: Summary The effects of recombinant human erythropoietin (rHuEpo) on megakaryocytopoiesis and platelet production were investigated in splenectomized, sham-operated and intact mice. When splenectomized mice were injected with 50 U of rHuEpo daily for 4 d, peripheral platelet counts, megakaryocyte (MK) size and the percentage of mature MK (stage IV MK) increased from 1 to 4 d after the initial rHuEpo injection. The total number of marrow MK colony forming units (CFU-MK) increased from 2 to 4 d after the initial injection. Furthermore, from days 6 to 8, the total number of marrow MK also increased. In addition, when the dose of rHuEpo was increased to 20 ˜ 200 U per mouse, clear dose responses were detected in platelet counts and MK numbers. When sham-operated and intact mice were injected with rHuEpo, no significant change in platelet numbers was detected. The number of MK, MK size and the number of CFU-MK increased only in spleen, not in bone marrow. Our data indicate that Epo has stimulatory effects on platelet production in vivo, if used in sufficient quantities.
British Journal of Haematology 03/2008; 76(2):260 - 268. · 4.94 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: High mobility group box 1 protein (HMGB1), originally described as a non-histone, DNA binding protein, was recently identified as a late mediator of inflammation via its extracellular release from activated macrophages/monocytes. In the present study, we report that intracellular HMGB1 synergizes with a macrophage/monocyte-specific E26 transformation-specific sequence (Ets) transcription factor PU.1 to transactivate the promoter of the IL1B gene coding a 31-kDa proIL-1beta protein. The -131 to +12 IL1B promoter, which possesses a PU.1 binding motif essential for its transactivation, was induced when HMGB1 expression vector was transfected into murine RAW264.7 macrophage cells. Our glutathione S-transferase-pulldown and coimmunoprecipitation assays demonstrated direct physical interaction of HMGB1 with PU.1. Deletion of the PU.1 winged helix-turn-helix DNA-binding domain inhibited the association of the two proteins. In electrophoretic mobility shift assay using recombinant PU.1 protein, a ternary complex of PU.1, HMGB1 and PU.1-binding element within the IL1B promoter was generated. The importance of PU.1 was further supported by our observation that induction of the IL1B promoter was obtained only after PU.1 expression in PU.1-deficient murine EL4 thymoma cells. Thus, our data raise the possibility of a novel mechanism which sustains and amplifies inflammatory reactions through physical interaction of PU.1 with intracellular HMGB1 in macrophages/monocytes.
European Journal Of Haematology 02/2008; 80(1):10-9. · 2.55 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Constitutive activation of nuclear factor (NF)-kappaB is a common feature of human T-cell leukemia virus type I (HTLV-I)-transformed T cells. Inhibition of NF-kappaB activity reduces cell growth and induces apoptosis of HTLV-I-transformed T cells, suggesting a central role of NF-kappaB in their proliferation and survival. In this study, we investigated whether MyD88, an adaptor protein of Toll-like receptor (TLR) signaling, contributes to constitutive NF-kappaB activation in HTLV-I-transformed T cells.
Activation status of MyD88 and interleukin (IL)-1R-associated kinase 1 (IRAK1) in HTLV-I-transformed human T cells, MT2, MT4, and HUT102 was examined by using Western blot and immunoprecipitation. TLR expression was evaluated with reverse transcription polymerase chain reaction. An expression vector encoding a dominant negative MyD88 with a deletion of its death domain (MyD88dn) was transfected into MT2 cells to evaluate roles of MyD88 in spontaneous activation of cytokine gene promoters and transcription factors, proliferation, and apoptosis in HTLV-I-transformed T cells.
Constitutive association of MyD88 with IRAK1 was observed in all three of HTLV-I-transformed T cells, but not in HTLV-I-negative T cells, such as Jurkat, HUT78, and MOLT4. MT2 cells showed expression of TLR-1, -6, and -10 mRNAs. Constitutive activation of NF-kappaB and NF-IL-6 and cytokine gene promoters, such as IL-1alpha, interferon-gamma, and tumor necrosis factor-alpha in MT2 cells was inhibited by MyD88dn expression. MyD88dn reduced proliferation and induced apoptosis of MT2 cells. HTLV-I Tax enhanced TLR expression and synergistically activated NF-kappaB with wild-type MyD88.
Our results show a novel pathway in NF-kappaB activation in HTLV-I-transformed T cells and further demonstrate a critical role of MyD88 in their dysregulated gene activation, survival, and proliferation.
[Show abstract][Hide abstract] ABSTRACT: Outpatient treatment in the cancer chemotherapy center was begun in April 2005 at the University of Occupational and Environmental Health Hospital. Drugs were prescribed 2,590 times during the past year. Times for intravenous drip for various regimens and outpatient chemotherapy desired by patients showed a rough. The number of incidents was three (0.12%) and no accidents occurred. There were 74 consultations with pharmacists about prescriptions (2.6%) and 286 (11.0%) with nurses. Both types of consultation decreased and their contents were different. The number of consultations about prescriptions by special staff at the cancer chemotherapy center was less than at other departments. Therefore, a system assuring safe management is critically required for the establishment of a system for outpatient cancer chemotherapy treatment.
Gan to kagaku ryoho. Cancer & chemotherapy 01/2008; 34(13):2259-62.
[Show abstract][Hide abstract] ABSTRACT: Rapid induction of transcription is known to be mediated by factors which bind DNA following post-translational modification. We report here that non-tyrosine phosphorylated (NTP)-Stat1 is involved in a cooperative interaction with Spi-1/PU.1 and IRF8 to form a pre-associated, poised complex for IL1B gene induction. A double point mutation at a putative STAT binding site, which overlaps this composite Spi-1 x IRF8 site located in the LPS and IL-1 response element (LILRE), inhibited human IL1B LPS-dependent reporter activity to about 10 percent of the control wild type vector. Chromatin immunoprecipitation revealed stimulation-independent constitutive binding of IRF8, Spi-1 and NTP-Stat1 at the LILRE, while binding of C/EBP beta was activated at an adjacent C/EBP beta site after LPS stimulation. In contrast to Stat1, IRF8 was tyrosine phosphorylated following LPS treatment. Supporting the involvement of NTP-Stat1, LPS-induced IL1B reporter activity in monocytes was enhanced by ectopic expression of NTP-Stat1 Y701F. In contrast, co-expression of a Y211F IRF8 mutein functioned as a dominant-negative inhibitor of LPS-induced IL1B reporter activity. In vitro DNA binding using extracts from LPS-treated monocytes confirmed that the LILRE enhancer constitutively binds a trimolecular complex containing IRF8, Spi-1 and NTP-Stat1. Binding studies using in vitro-expressed proteins revealed that NTP-Stat1 enhanced the binding of Spi-1 and IRF8 to the LILRE. Co-expression of TRAF6, an LPS surrogate, with Spi-1 and IRF8 enhanced IL1B reporter activity in HEK293R cells, which was dramatically reduced when Y211F IRF8 was co-expressed. These results suggest that the rapid transcriptional induction of an important inflammatory gene is dependent upon constitutive cooperative binding of a Spi-1 x IRF8 x NTP-Stat1 complex to the LILRE, which primes the gene for immediate induction following IRF8 phosphorylation. Phosphorylation of chromatin pre-associated factors like IRF8 may be an important strategy for the rapid transcriptional activation of genes involved in innate immunity.
[Show abstract][Hide abstract] ABSTRACT: The purpose of this study was to establish criteria to predict the need for emergency hospitalization of patients receiving chemotherapy, based on information at presentation. 158 consecutive patients treated for cancer at the Cancer Chemotherapy Center in the University of Occupational and Environmental Health were examined. The number of emergency hospitalization cases for outpatients undergoing cancer chemotherapy was 14 (8.9%) and including seven lung carcinomas, six hematological carcinomas, and one mediastinal tumor. The reason for emergency hospitalization in twelve (85.7%) of the cases was infection. No significant difference was observed between the cases with and without emergency hospitalization regarding age, gender, cancer type, previous treatment, objective of the chemotherapy, or line of chemotherapy. A significantly higher number of the emergency cases were associated with performance status 2, severe adverse events and comorbidity than with a performance status 0-1 where there were no or only mild adverse events and no comorbidity. Multiple logistic regression models indicated that severe adverse events and comorbidities were independent predictive factors for patients with emergency hospitalization. By combining selected clinical information for outpatients receiving cancer chemotherapy, the need for emergency hospitalization could be predicted.
Anticancer research 01/2007; 27(2):1133-6. · 1.71 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A quality of life (QOL) assessment has become increasingly common in cancer clinical trials. Seventy-four consecutive patients treated for cancer between August 2005 and January 2006 at the Cancer Chemotherapy Center in the University of Occupational and Environmental Health, Japan, were examined. The 8-Short form health survey (SF-8) was utilized as a comprehensive scale and quality of life questionnaire for cancer patients treated with anticancer drugs (QOL-ACD) as disease specific scale for the QOL evaluation. The QOL for outpatients was investigated in comparison with that for inpatients. All questionnaires were collected and baseline questionnaires were filled in by 98.1% of the subjects. The physical comprehensive score (PCS) of SF-8 for the outpatients was higher than that for the inpatients. The physical condition of the outpatients was better than that of inpatients. There was no difference in the baseline scores of the QOL-ACD scales in daily activity, psychological condition, social attitude, and face scale of the analyzed domains between the two groups. Furthermore, a longitudinal study from admission to outpatient was carried out on 27 patients who were treated on an outpatient basis in our clinic. No difference in the baseline scores of the SF-8 and QOL-ACD scales were observed in any of the analyzed domains. These data suggest that the present QOL study has a sufficient feasibility for the outpatients evaluated in our study, and QOL of outpatients after discharge is equal to that of inpatients receiving cancer chemotherapy.
Anticancer research 01/2007; 27(2):1127-32. · 1.71 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We conducted a questionnaire survey to evaluate patients at the cancer chemotherapy center of the university of occupational and environmental health hospital from January to March 2006. Thirty-three (61.1%) and 5 (9.2%) of 54 patients wished to receive chemotherapy on an outpatient basis and as inpatients, respectively. Among the reasons for wishing to be treated as outpatients were the following: Keeping in touch with family, not wanting to be hospitalized, a desire to work, and the wish to continue hobbies, at 45.5 (15/33), 36.4 (12/33), 24.2 (8/33), and 24.2 (8/33), respectively. Twenty-four (44.4%) patients voiced concerns about chemotherapy at an outpatient clinic in terms of associated adverse events, feeling bad about causing trouble to their family, and the desire to have consultations at night or on holidays. Thirty-eight (70.4%) patients wished to have chemotherapy within 3 hours, and 42 (77.7%) of them requested improvements in the examinations, time, duration of chemotherapy, and the effect of chemotherapy. These findings indicate that a major part of patients wish to receive cancer chemotherapy at an outpatient clinic within 3 hours. However, they also have serious reservations about any associate adverse events. We therefore need to improve the overall environment for cancer chemotherapy patients in order to allow them to feel safe when undergoing such treatments.
Gan to kagaku ryoho. Cancer & chemotherapy 12/2006; 33(11):1681-3.