bluuebunny/crossref_metadata_2025_split huggingface dataset:
siRNA Targeting bcl-2 Gene Improve H and RA Drug-Sensitivity in HL-60 Cells.
Authors: Hu Haiyan , Zhang Yuan , Kunyuan Guo 
Published: April 2020

Abstract
The anti-apoptosis gene, bcl-2, is also related to drug-resistance, which expresses markedly higher on all kinds of leukemia. small interference RNA can silence the targeted mRNA expression.
Objective: To study the effect of G3139 and siRNA targeting bcl-2 gene which increase the sensitivity of HL-60 cell to HT, Ara-C, As2O3 and RA.
Method:Incubate leukima cell HL-60 with H, Ara-C, As2O3 and RA, meanwhile transfected siRNAs or G3139 in it by lipofectamine2000 with OPTI-MEM cultural medium for 6h, then add equal volume RPMI-1640 culture medium containing 20% inactive newborn calf serum. After transfection for 24,48 and 72h, cell proliferations were determined by MTT method. After transfection for 48h, we test the level of Bcl-2 protein, ROS and membrane potential of mitochondrion in HL-60 cell by FCM.
Result:
Compare with control group, the G3139 and siRNA targeting bcl-2 combine with H or Ara-C or As2O3 have significant effects on inhibiting the proliferation of HL-60(P &lt;0.05); After transfecting 48h, the level of Bcl-2 protein and ROS decrease significantly, meanwhile membrane potential of mitochondrion in HL-60 cells increase markedly (P&lt;0.01). Compare with control group, the G3139 and siRNA targeting bcl-2 combine with RA have significant effects on inhibiting the proliferation of HL-60 and the level of Bcl-2 protein decrease, but there were not difference between test groups and control group on the level of ROS and membrane potential of mitochondrion (P&lt;0.01).
Conclusion: The effective siRNA2, targeting bcl-2, can increased the drug-sensitivity though inhibiting the expression of Bcl-2 protein. Different drug has different mechanism to induce the leukimic cell apoptosis.
The Requirement of Reactive Oxygen Species Generation in the Apoptosis of Leukemia Cells Induced by 2-Methoxyestradiol.
Authors: Kunyuan Guo , She Miaorong , Hu Haiyan , Niu Xinqing , Tu Sanfang , Lu Xiaoxun 
Published: April 2020

Abstract
2-Methoxyestradiol (2-ME) is a new anticancer agent currently under investigation for treatment of leukemia. We evaluated the effects of 2-ME-induced apoptosis in two myeloid leukemia cell lines (U937 and HL-60) in association with reactive oxygen species (ROS) generation. We found that 2-ME resulted in viability decrease in a dose-dependent manner, generated ROS: nitric oxide and superoxide anions, and mitochondria damage. 2-ME-induced apoptosis correlated with increase in ROS. Quenching of ROS with N-acetyl-L-cysteine protected leukemia cells from the cytotoxicity of 2-ME and prevented apoptosis induction by 2-ME. Furthermore, addition of manumycin, a farnesyltransferase inhibitor, demonstrated by our previous studies that induced apoptosis of leukemic cells and induced ROS, significantly enhanced the apoptosis-induced by 2-ME. In conclusion, cellular ROS generation play an important role in the cytotoxic effect of 2-ME. It is possible to use ROS-generation agents such as manumycin to enhance the antileukemic effect. Such a combination strategy need the further in vivo justify and may have potential clinical application.
Functional CXCR4 Expressing Microparticles and SDF-1 Correlate with Circulating AML Cell Counts.
Authors: Kalinkovich Alexander , Tavor Sigal , Avigdor Abraham , Kahn Joy , Brill Alexander , Petit Isabelle , Goichberg Polina , Tesio Melania , Netzer Neta , Naparstek Elizabeth , Hardan Izhar , Nagler Arnon , Resnick Igor , Tsimanis Alexander , Lapidot Tsvee 
Published: April 2020

Abstract
Stromal cell-derived factor-1 (SDF-1/CXCL12) and its receptor CXCR4 are implicated in the pathogenesis and prognosis of AML. Cellular microparticles (MPs), the submicron vesicles shed from the plasma membrane of various circulating cells, are associated with numerous human disorders. In the present study, we studied the putative relationships between CXCR4/SDF-1 axis and MPs in AML. We detected CXCR4 expressing MPs (CXCR4+MPs) in the peripheral blood and bone marrow plasma samples of normal donors (n=24) and newly diagnosed adult AML patients (n=26). The majority of CXCR4+MPs in AML patients were CD45+ whereas in normal individuals they were mostly CD41+. In samples from AML patients, the levels of CXCR4+MPs and total SDF-1 were significantly elevated as compared to normal individuals. Importantly, we found a strong correlation between the levels of CXCR4+MP and white blood cell (WBC) counts in the peripheral blood and bone marrow plasma obtained from the AML patients. Of interest, functional, non-cleaved SDF-1 levels were reduced in these patients compared to normal individuals, and also strongly correlated with the WBC counts. Furthermore, our data indicate N-terminal truncation of the CXCR4 molecule in the MPs of AML patients. This was found also in MPs obtained from the conditioned media of normal human CD34+ progenitors lentiviarlly transduced with CXCR4 vector in vitro. Appearance of MPs possessing N-terminally truncated CXCR4 in AML patients is likely to be dependent on proteolytic enzymes, such as elastase, which was elevated. However, MPs isolated from AML patients were capable of transferring functional CXCR4 molecule to the AML-derived HL-60 cells, enhancing their migration to SDF-1 in vitro and increasing their homing to the bone marrow of irradiated NOD/SCID mice. The CXCR4 antagonist AMD3100 reduced the increased migration and homing of MP treated HL-60 cells. Taken together, these findings suggest that functional CXCR4+MPs and SDF-1 are involved in the progression of AML. We propose that their levels are potentially valuable as an additional diagnostic AML parameter. Moreover, our findings suggest also the need for CXCR4- and SDF-1-target therapeutic approaches, clinically relevant in AML in the near future.
Epigenetic-Based Treatment Induces Apoptosis in Leukemic Cell Lines.
Authors: Carnicer Maria J. , Lasa Adriana , Serrano Elena , Pena Jorge , Remacha Angel F. , Aventin Anna M. , Nomdedeu Josep F. 
Published: April 2020

Abstract
Pharmacological treatment of cancer cells with demethylating agents and histone deacetylase inhibitors synergistically reactivates the transcription of previously silenced genes.
The aim of this study was to investigate the antileukemic properties of a DNA mehtyltransferase inhibitor 5-aza-2′-deoxycytidine (5-Aza-dC),an histone deacetylase inhibitor Trichostatin A (TSA) and trans-retinoc acid (ATRA), alone or in combination. The effects of these drugs on apoptosis, cell cycle progression, cell-survival pathways and restoration of proliferation-associated genes silenced by aberrant epigenetic mechanisms were assessed. Four human leukemic cell lines were used: lymphoid cell-lines REH (Tel-Aml1+) and DAUDI (c-myc) and myeloid cell-lines Kasumi-1(Aml1-Eto+) and NB4(Pml-Rara+). The cells were cultured for three days. Cell viability, percentage of apoptosis, cell cycle and apoptosis-controlling proteins were examined by multiparametric flow cytometry, western blotting and a RT-PCR low-density array (LDA)containing 48 probes. Combined 5-Aza-dC and TSA treatment induced a high degree of apoptosis and affected the cell-cycle in all the cell lines analyzed. Increased levels of cleaved PARP were detected after the double treatment and paralleled those of caspase-3 except for DAUDI cell-line where there was no cleavage of procaspase 3 suggesting that this treatment induced a death-pathway independent of caspase-3 activation. Furthermore, ATRA alone has a limited capacity to induce apoptosis in all the tested cell-lines.
The genes upregulated following the combined epigenetic-active treatment in DAUDI were; PRKCG, TNFSF10, TNFRSF10B, MAPK8, CASP9, APAF1, PER1, CDKN1A, CCND1. In the REH cell line were; PRKCG, TNFRSF10A, TNFRSF10B, MAPK8, CASP8, CASP9, APAF1, CASP3, AKT1, PIK3CG, BIK, PRKCG, MCL1, CCND1, CCND2, EZH2, PER1, GCLC, GSTP1. In the KASUMI-1 cell line were; PRKCABP, TNFSF10, TNFRSF10A, TNFRSF10B, FAS, CFLAR, CASP8, APAF1, CEBPA, CCND1, CDC25C, CDKN1A, EZH2.For the NB4 cell-line were: PRKCG, PRKCABP, AKT1, TNFSF10, CFLAR, CASP8 and CASP9. We can conclude that combined treatment with demethylating agents and histone deacetylase inhibitors may be active in a wide range of human leukemias. The potential use of this combination in the commonest form of pediatric ALL warrants further investigation.
Synergy Studies between the Nitric Oxide (NO) Donor JS-K and Other Anti-Leukemic Agents.
Authors: Shami Paul J. , Eddington Jordan K. , Udupi Vidya , Kosak Ken M. , Saavedra Joseph E. , Keefer Larry K. 
Published: April 2020

Abstract
NO induces apoptosis in Acute Myelogenous Leukemia (AML) cells. Glutathione S-Transferases (GST) play an important role in multidrug resistance and are upregulated in 90% of AML cells. We have designed a prodrug class that releases NO on metabolism by GST. O2-(2,4-Dinitrophenyl) 1-[(4-ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diolate (JS-K, a member of this class) has potent antileukemic activity (Molecular Cancer Therapeutics 2:409–417,2003). We sought to identify synergistic interactions between JS-K and other antileukemic agents. The human HL-60 myeloid leukemia cell line was used for in vitro studies of the combination of JS-K with Daunorubicin (Dauno), Cytarabine (Ara-C) or Etoposide (Etop). Drugs were added to cells in logarithmic growth. Cell viability was determined 3 days later using the CellTiter 96 assay (Promega, Madison, WI). The median effect method of Chou and Talalay was used to determine synergistic, antagonistic, or additive effects with the CalcuSyn software (Biosoft, Ferguson, MO). Combinations of JS-K added simultaneously, 2 hours before or 2 hours after the other compounds were used. Drug combinations were added in increments at a fixed concentration ratio. The 50% growth inhibitory concentrations (IC50) for Dauno, Ara-C, Etop, and JS-K alone were 0.07, 0.18, 0.79, and 0.30 μM, respectively. JS-K and Dauno (Dauno:JS-K fixed concentration ratio = 1:5) were antagonistic in all 3 drug sequences. The Combination Index (CI) at the 50% fraction affected was 12.5 ± 1.6, 10.7 ± 0.41, and 7.2 ± 0.46 for simultaneous, JS-K first and Dauno first additions, respectively. JS-K and Etop (Etop:JS-K fixed concentration ratio = 2:1) were antagonistic but to a lesser degree. The CI at the 50% fraction affected was 3.03 ± 0.26, 2.2 ± 0.33, and 1.7 ± 0.082 for simultaneous, JS-K first and Etop first additions, respectively. JS-K and Ara-C (Ara-C:JS-K fixed concentration ratio = 1:1) showed strong synergy. The CI at the 50% fraction affected was 0.37 ± 0.23, 0.24 ± 0.27, and 0.15 ± 0.11 for simultaneous, JS-K first and Ara-C first additions, respectively. We conclude that Ara-C and JS-K have synergistic anti-leukemic activity and warrant further exploration in combination.
The Transcription Factor ZBP-89 Controls Generation of the Hematopoietic Lineage in Zebrafish and Mouse Embryonic Stem Cells.
Authors: Li Xiangen , Xiong Jing-wei , Shelley Carl Simon , Park HeiYoung , Arnaout M. Amin 
Published: April 2020

Abstract
Hematopoietic development is closely linked to that of blood vessels and the two processes are regulated in large part by transcription factors that control cell fate decisions and cellular differentiation. Both blood and blood vessels derive from a common progenitor, termed the hemangioblast, but the factors specifying development and differentiation of this stem cell population into the hematopoietic and vascular lineages remain ill defined. Here we report that knockdown of the Krüppel-like zinc finger ZBP-89 (ZNF148) in zebrafish embryos results in a bloodless phenotype, caused by disruption of both primitive and definitive hematopoiesis, leaving primary blood vessel formation intact. Expression of the primitive (Scl, Lmo2, Gata2, Gata1) and definitive (Runx1, c-Myb) hematopoietic stem cell lineage markers in ZBP-89 morphants were significantly decreased while expression of the lymphopoiesis lineage markers Ikaros and Gata3 was unaffected. These developmental HSC defects are rescued by injection of human ZBP-89 RNA, SCL RNA or ZBP-89 cDNA under control of the zebrafish Flk1 promoter. Injection of ZBP-89 RNA into cloche zebrafish embryos, which lack the hematopoietic and endothelial lineages, rescued hematopoiesis but not vasculogenesis. Overexpression of ZBP-89 in wild type zebrafish expanded hematopoietic progenitors (Scl+, Lmo2+, Gata1+) but reduced the endothelial cell markers FLK1 and TIE1. Forced expression of ZBP-89 in mouse embryonic stem cell cultures also induced the expansion of hematopoietic progenitors (c-KIT+, SCA1+) and definitive hematopoietic cell population (CD45+), but inhibited sprouting angiogenesis and the CD31+, VE-Cadherin+ endothelial cell population. These findings establish a unique regulatory role for ZBP-89, positioned at the interface between early blood and blood vessel development.
The Study of the Expression of TNF in Bone Marrow Specimens and Survival in Patients with Acute Myelogenous Leukemia.
Authors: Shum Merrill K. , Hageboutros Karine , Skaff Ziad , Kankipati Shoba , Hageboutros Alexandre 
Published: April 2020

Abstract
Background: The objective of the current study was to assess the correlation between the expression of TNF from bone marrow specimens and survival in patients with acute myelogenous leukemia (AML).
Methods: Immunohistochemical analysis of bone marrow biopsies from 10 patients recently diagnosed with AML was performed using the dakocytomation LSAB + system -HRP kit, to stain for TNF binding. TNF-α and TNF-β expression were analyzed for color intensity and assigned a score from (0–5+) with 5+ being bright and over-expressed.
Results: Bone marrow specimens from 4 women and 6 men (n=10), median age 34 (range 19–61 years), all diagnosed with AML, revealed high TNF-β expression (3, 4, 5+) in 8/10 patients and high TNF-α expression (3, 4, 5+) in 6/10 patients. Median survival was 5.5 (range 1–54 months). Fisher’s exact tests for paired samples showed no correlation between predictors (age, sex, survival) and outcomes (TNF-α, TNF-β). Pearson Chi-square tests showed no correlation between predictors (age, sex, survival) and outcomes (high TNF-α / high TNF-β expression, mixed TNF-α /TNF-β expression, low TNF-α / low TNF-β expression) [See chart below]. However there was an association seen with younger males expressing both high TNF-α and high TNF-β and older women expressing low TNF-α and low TNF-β. Interestingly, the eldest patient who expressed TNF-β 4+ and TNF-α 3+ also survived the longest. He achieved clinical remission after standard induction chemotherapy, but relapsed 8 times over a course of 54 months. During each clinical remission, the patient was treated with infliximab (a TNF-α blocker) for his Crohn’s disease.
Conclusion: Level of expression of TNF-α and TNF-β of bone marrow biopsies from our AML patients showed no correlation with age, sex or overall survival given the small sample size. A large, prospective trial will be needed to determine the significance of TNF-α or TNF-β expression with survival and whether TNF-α blockade alone or in combination with chemotherapy plays a role in the treatment of patients with AML over-expressing TNF-α.
SUMMARY OF ASSOCIATIONS OUTCOMES TNF α+TNFβ* TNF α** TNF β** *Pearson Chi-square with 2dF **Fisher’s Exact Test Sex 0.43 0.57 0.47 PREDICTORS Survival 1.00 1.00 1.00 Age 0.26 0.52 0.42
HLA-DRB1 Analysis in Hairy B-Cell Lymphoproliferative Disorder (HBLD): A Japanese Type of Benign Persistent Polyclonal B-Cell Lymphocytosis.
Authors: Nakahashi Hirotaka , Karasawa Masamitsu , Hashimoto Yoko , Irisawa Hiroyuki , Yokohama Akihiko , Saitoh Takayuki , Handa Hiroshi , Matsushima Takafumi , Murakami Hirokazu , Tsukamoto Norifumi , Nojima Yosihisa 
Published: April 2020

Abstract
Two types of benign persistent polyclonal B-cell Lymphocytosis have been reported: persistent polyclonal B-cell lymphocytosis (PPBL) in Western countries and hairy B-cell lymphoproliferative disorder (HBLD) in Japan. PPBL is characterized by B lymphocytosis with binucleated lymphocytes, polyclonal IgM, isolated splenomegaly, female sex, cigarette smoking and the presence of HLA-DR7 allele. Most patients with HBLD have atypical lymphocytes with microvilli and prominent membranous ruffles, serum polyclonal IgG, and splenomegaly. Only 9 patients with HBLD have been reported to date, and all of these patients are Japanese. Here, we report three new cases of HBLD. We performed HLA-DRB1 subtyping since the association between HBLD and HLA-DR4 has been suggested: 4 of 5 HBLD patients examined demonstrated the HLA allele. We also analyzed HLA-DRB1 alleles in a patient with hairy cell leukemia (HCL) and 3 with HCL Japanese variant (HCL-JV) to examine the possibility that polyclonal cell expansion in HBLD may be followed by transformation to these lymphoproliferative disorders. If the transformation were to occur, the patients should share the same HLA-DR allele.
Expanded B-lymphocytes in all HBLD cases displayed CD5− CD10− CD11c+ CD19+ CD20+ CD23− CD25− and CD103− cell surface phenotype and all patients exhibited a polyclonal increase in serum IgG. Phase-contrast microscopic and scanning electron microscopic examinations demonstrated lymphoid cells with many long surface microvilli. Analysis of immunoglobulin VH gene rearrangements showed a polyclonal pattern. Therefore, we diagnosed these patients as HBLD. HLA-DRB1 subtype analysis strengthened the close relation between HBLD and HLA-DR4: 2 patients having HLA-DRB1*0405 and one HLA-DRB1*0407. In the meanwhile one each patient with HCL and HCL-JV had HLA-DRB1*0405 allele.
Although the exact mechanism underlying the association between HBLD and HLAB1*04 needs to be elucidated, the presence of this HLA might be useful to establish correct diagnosis of the disease, and thus avoid patient exposure to unnecessary treatment. One each patient with HCL and HCL-JV has HLA-DRB1*0405, but a definitive conclusion cannot yet be drawn due to high HLA-DRB1*04 frequency in Japanese population (25.4%).
Significance of Marrow Angiogenesis Factors in Patients with Acute Myelogenous Leukemia.
Authors: Lin Liang-In , Lee Cheng-Yeh , Hu Chung-Yi , Tang Jih-Luh , Tien Hwei-Fang 
Published: April 2020

Abstract
Bone marrow (BM) neoangiogenesis plays an important role in the pathogenesis of acute myelogenous leukemia (AML). Paracrine exchange of growth factors among AML blasts, endothelial cells, and other marrow compartments is believed to contribute to the pathogenesis of AML. The vascular endothelial growth factor (VEGF) and the angiopoietin (ANG) family are the two major classes of growth factors associated with angiogenesis. In the present study, BM plasma from 52 AML patients and 20 normal donors were investigated. We measured the marrow concentrations of seven molecules associated with angiogenesis, VEGF, VEGF/PlGF, VEGF-C, VEGF-D, ANG-1, ANG-2, and Tie-2, by using enzyme-linked immunosorbent assay (ELISA). Compared to normal donors, the marrow levels of VEGF/PlGF, ANG-2, and Tie-2 were increased in patients with AML (p&lt;0.005, &lt;0.0001 and &lt;0.0001, respectively). On the contrary, VEGF-C and ANG-1 levels were decreased in patients with AML (p&lt;0.0001 and &lt;0.0005, respectively). Although the values were not completely normalized, sequential study revealed that at complete remission (CR) period, VEGF/PlGF, ANG-2, and Tie-2 levels were decreased (p&lt;0.005, &lt;0.005 and =0.02, respectively), while VEGF-C increased (p=0.04). Kaplan-Meier survival curves showed that the subgroup of patients with lower Tie-2 level (29 ng/ml) had a longer median survival time than those with higher Tie-2 level (18.9 months versus 2.5 months, p&lt;0.005). In addition, subgroups of patients with higher VEGF/PlGF level (1 pg/ml) and VEGF-D level (350 pg/ml) also had a longer median survival time than those with lower levels (20.8 months versus 7.2 months, p=0.03 and 18.9 months versus 3.4 months, p=0.03, respectively). Taken together, there were significant differences in VEGF/PlGF, VEGF-C, ANG-1, ANG-2, and Tie-2 expressions between AML patients and normal donors. Expressions of VEGF/PlGF, VEGF-D, and Tie-2 might be prognostic markers in AML patients.
Prognostic Significance of FLT3 Mutations in Paired Initial and Relapse Marrow Samples of Acute Myeloid Leukemia Excluding Acute Promyelocytic Leukemia.
Authors: Cho Young-Uk , Chi Hyun-Sook , Jang Seongsoo , Park Chan-Jeoung , Lee Je-Hwan , Lee Kyoo Hyung , Lee Jung-Hee , Lim Ho-Jun , Seo Jong-Jin 
Published: April 2020

Abstract
Fms-Like tyrosine kinase 3 (FLT3) mutations are related to poor prognosis in acute myeloid leukemia (AML), with the exception of acute promyelocytic leukemia (APL), in which the prognostic significance of this mutation is not firmly established even though it is more frequent than other FAB subtypes. We investigated FLT3 internal tandem duplications (FLT3/ITD) and/or D835 point mutations in both initial and relapse marrow samples from 120 AML patients, excluding APL. FLT3/ITDs were found in 18 initial and 16 relapse samples (15.0 and 13.3%, respectively). D835 mutations were found in one initial sample and 4 relapse samples. Two cases acquired FLT3/ITD at relapse, 4 cases lost mutations at relapse, and one had a different allele of FLT3/ITD at relapse. Patients who had FLT3/ITDs at initial diagnosis showed shorter median duration from diagnosis to relapse and shorter overall survival compared with those without mutations (median duration to relapse: 8.3 and 11.0 months, respectively, P = 0.074; overall survival: 12.2 and 20.8 months, respectively, P = 0.058). These findings were more pronounced when the FLT3/ITDs were acquired at relapse or discrepant patterns were noted between diagnosis and relapse (median duration to relapse: P = 0.047 and 0.044, respectively; overall survival: P = 0.002 and 0.038, respectively). In conclusion, the present study showed that patients with AML had heterogeneous patterns of FLT3/ITD or D835 mutations, either acquisition or loss of the mutations at relapse. The overall survival and relapse-free survival times were significantly shorter in the mutation acquisition group than in those with mutation loss at relapse.