bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development as Anti-Leukemic Agents, a Novel Synthesis of Phospha Sugar Derivatives in Therapy for Leukemias, and Analysis of Their Characterizations.
Authors: Nakamura Satoki , Okinaka Keiji , Hirano Isao , Ono Takaaki , Shigeno Kazuyuki , Shinjo Kaori , Fujie Michio , Iguchi Yukiko , Niimi Taishi , Ito Satoru , Suyama Takuya , Asai Kazuhide , Yamashita Mitsuji , Ohnishi Kazunori 
Published: October 2019

Abstract
[Background] Sugar derivatives whose oxygen atom in the hemiacetal ring is replaced by a carbon, nitrogen, sulfur atom, and etc., are celled as pseudo sugars. The synthesis and bioassay of these pseudo sugars are well investigated and are known as potential bioactive materials. Phospha sugar, which is one of pseudo sugars, having a phosphorus atom instead of the oxygen atom in the hemiacetal ring, is little known. The synthesis and bioassay of phospha sugars are not well studied in detail. We synthesized the 2, 3, 4-tribromo-3-methyl-1-phenyl phospholane 1-oxide (TMPP) and 2, 3-dibromo-3-methyl-1-phenyl phospholane 1-oxide (DMPP) by the nucleophilic substitution reactions, and found their anti-leukemic activities.
[Purpose] The aims of the present study were to evaluate the inhibition of proliferation and induction of apoptosis in leukemia cell treated with TMPP or DMPP, and define the target molecules for TMPP in leukemia cells.
[Methods] The cells used in this study were human leukemia cell lines, K562, U937, and YRK2 cells. For proliferation analysis, MTT assays were performed in leukemia cells treated with TMPP. For cell cycle analysis, flow cytometory analysis was performed in leukemia cells treated with TMPP or DMPP by PI staining. For analysis of mitotic regulatory proteins (p27, p21, Skp2, Cdc25B, Cyclin D1, Survivin, and Aurora kinase B), Western blotting was performed in leukemia cells treated with TMPP.
[Results] In leukemia cell lines, DMPP and TMPP significantly inhibited the cell proliferation, and TMPP more strongly inhibited the cell proliferation than DMPP. 10 μM TMPP significantly induced G2/M phase arrest, and 25 μM TMPP induced apoptosis in leukemia cells. In leukemia cells, 10 μM TMPP reduced protein of Aurora kinase B, Survivin, Cyclin D1, Skp2 KIS, and FoxM1, while increased protein expression of p21and p27 by western blot analysis. Moreover, 25 μM TMPP activated caspase-3 and caspase-9, cleaved PARP, and reduced Bcl-2.
[Conclusions] In this study, we report in the first time the possibility of phospha sugar derivatives as anti-leukemic agents in therapy for leukemias, and analysis of their characterizations. DMPP significantly inhibited the proliferation, and induced apoptosis of leukemia cells. These results demonstrated that the FoxM1, which is an essential transcription factor for cell growth and regulates expression of the mitotic regulators, is one of the targets for DMPP. Therefore, DMPP or TMPP has possibility as new agents for FoxM1 in leukemia therapy.
Reversal of P-Glycoprotein-Dependent Resistance to Adriamycin by 5-bromotetrandrine in K562/A02 Cell Line.
Authors: Chen Bao-An , Wang Jue-Qiong , Ding Jia-Hua , Gao Feng , Chen Jian , Gao Chong , Sun Yun-Yu , Zhao Gang , Chen Ning-Na , Pei Xiao-Ping , Dai Yong-Yuan , Shun Qian 
Published: October 2019

Abstract
Objective: The present study aimed to evaluate the MDR reversal activity of bromotetrandrine (BrTet), a bromized derivative of tetrandrine (Tet), in vitro.
Methods: Drug sensitivity was determined using the MTT assay. Adriamycin (ADM) accumulation, the protein levels of P-glycoprotein (P-gp) and the apoptotic changes were analyzed by fluorospectrophotometry, respectively. The mRNA levels of P-gp was determined by RT-PCR.
Results: BrTet at 0.25, 0.5 and reversed ADM resistance in MDR K562/A02 cells dose-dependently and its potency was greater than that of Tet at the same concentrations. The IC50 of ADM for K562 and K562/A02 cells were 55.122 mg/l and 1.1373 mg/l, respectively. Treating K562/A02 cells with BrTet(1uM)and TTD(1uM)both for 48 hours partially restored the sensitivity of K562/A02 cells to ADM (IC50 were 4.7729 mg/l and 13.584 mg/l respectively) but had not effect on K562 cells. The fold reversal (FR) were 11.55 and 4.06 respectively. K562/A02 cells showed apoptotic characteristics after treated with Brtet and Tet both for 48 hours compared with control group(apoptosis rate was 61.1%, 11.1% and 9.9%,respectively); Fluorospectrophotometric assay showed that BrTet significantly increased the intracellular accumulation of ADM in K562/A02 cells in a dose-dependent manner. The fluorescence intensity of intracellelar ADM in K562/A02 cells treated with ADM(1mg/L)was 33% of that in K562 cells. BrTet and Tet elevated the intracellular ADM concentration in K562/A02 cells up to 52% and 69%,respectively. BrTet also inhibited the overexpression of P-gp in K562/A02 cells. The fluorescence intensity of P-gp in K562 and K562/A02 cells was 0.5 and 97.97.The P-gp expression was down after treated with BrTet and TTD (65.05 and 54.86). The mdr1 mRNA was also down regulated.
Conclusions: BrTet showed significant MDR reversal activity in vitro. Its activity may be related to the inhibition of P-gp overexpression and the increase in intracellular accumulation of anticancer drugs. BrTet may be a promising MDR modulator for eventual assessment in the clinic.
Difference of Drug-Resistance between Single-Factor Resistance Cell Line K562/MDR1 Transfected with mdr1 Gene and Multi-Factor Resistance Cell Line K562/A02 Induced by Doxorubicin.
Authors: Gong Yuping , Ye Xueshi , Liu Ting 
Published: October 2019

Abstract
Objective To study difference of drug-resistance between single-factor resistance cell line K562/MDR1 transfected with mdr1 gene and multi-factor resistance cell line K562/A02 induced by doxorubicin.
Methods Retroviral virions carrying the complete sequence of mdr1 gene cDNA were produced and infected drug-sensitive leukemia cell line K562 and mdr1 single-factor resistance cell line K562/MDR1 was established. The difference of drug-resistance between K562/MDR1 and K562/A02, a kind of multi-factor resistance cell line induced by doxorubicin, was studied by checking the expression of mdr1 gene and Pgp, daunorubicin efflux rate, MTT drug sensitivity to chemotherapeutic drug. Lentiviral vector encoding shRNA which targeted MDR1 gene was transfected into two kinds of cell lines and effect of RNAi on reversing drug resistance was detected.
Results The results of Q-PCR and flow cytometry demonstrated that there were high expression of mdr1 mRNA and Pgp in both kinds of drug-resistance cell lines and no difference between them. The function of Pgp detected by daunorubicin efflux rate is higher in K562/MDR1 (90.93%) than K562/A02 (78.67%). The results of MTT test showed that IC50 of K562/MDR1 and K562/A02 is 0.55 and 1.22μmol/L respectively and this confirmed that drug-resistance in K562/A02 is higher than that in K562/MDR1. After RNA interference, the expression of the mdr1 gene and Pgp in K562/MDR1 markedly was down-regulated and the drug resistance was restored and IC50 is 0.16μmol/L, similar to K562 sensitive cell line. The expression of the mdr1 gene and Pgp in K562/A02 markedly was downregulated too, and drug resistance to anticancer drug is reduced to some extent but IC50 is 0.56μmol/L, it is still higher than that in sensitive cell line.
Conclusion Drug-resistance in K562/A02 induced by anticancer-drug was made of many factors and it is more resistance to anticancer-drug than that in K562/MDR1 caused by mdr1 gene. Due to only mdr1 resistance, K562/MDR1 is better cell model to make mdr1/Pgp research.
Addition of PTK787/ZK 222584 Can Lower the Dosage of Amsacrine To Achieve Equal Amounts of AML Cell Death.
Authors: Weidenaar Alida C. , de Jonge Hendrik J.M. , Fidler Vaclav , Elst Arja ter , Meeuwsen Tiny , Douwes Jenny , Bouma Jessica C.A. , Hählen Karel , Kamps Willem A. , de Bont Evelina S. 
Published: October 2019

Abstract
Acute myeloid leukemia (AML) is a disease with a poor prognosis. At this moment conventional chemotherapy is the gold standard for the treatment of AML. It is demonstrated that AML cells express VEGFA and VEGFC as well as KDR (VEGFR2), the main receptor for downstream effects, resulting in an autocrine pathway for cell survival. The present study investigates the role of PTK787/ZK 222584, a potent inhibitor of VEGF receptor tyrosine kinases, upon leukemic cell death, and the possibility of an additional effect upon cell death achieved by a conventional chemotherapeutic drug Amsacrine. In 3 AML-cell lines and 33 pediatric AML patient samples we performed total cell kill assays to determine the percentage of cell death achieved by PTK787/ZK 222584 (5–100 μM) and/or Amsacrine (0.001–2 μg/ml). Both drugs induced AML cell death. The LC50 values (drug concentration needed to kill 50% of the leukemic cells) for HL-60, TF-1 and THP-1 were 27 μM, 49 μM and 24 μM respectively. An excellent survival was seen on a KDR-negative cell line demonstrating that the cell death induced by PTK787/ZK 222584 is not a toxic effect. The primary blasts were overall 5–10 times more sensitive to PTK787/ZK 222584 than the cell lines, with a median LC50 value of 5.1 μM. Patient samples with a LC50 value below the median did not differ from patients above the median regarding age, sex, FAB classification or WBC count. With a response surface analysis we investigated the additional effect of PTK787/ZK 222584 upon cell death achieved by Amsacrine; we estimated the concentration combinations resulting in the same cell survival. We could show that, in cell lines as well as in primary AML blasts, the concentration of Amsacrine can be lowered and replaced for a certain dose of the potentially less toxic VEGFR inhibitor to achieve the same percentage of leukemic cell death. This study shows that PTK787/ZK 222584 might have more clinical potential in AML when combined with chemotherapy, e.g. Amsacrine. In addition, it is known from literature that topoisomerase inhibitors, e.g. Amsacrine, can cause treatment related AMLs. Moreover, many survivors of childhood or adolescent cancer experience treatment related cardiovascular complications that can impair their quality of life years after treatment. Therefore, reduction of complications and long-term effects of chemotherapy is warranted and might be achieved by lowering dosages of Amsacrine and replace it by other drugs with a lower toxicity profile, such as PTK787/ZK 222584.
Integrin Linked Kinase Is Important in Platelet Signalling and Function.
Authors: Tucker Katherine L. , Stevens Joanne M. , Jordan Peter A. , Sage Tanya , Jones Sarah , Barrett Natasha E. , St-Arnaud Rene , Frampton Jonathan , Dedhar Shoukat , Gibbins Jonathan M. 
Published: October 2019

Abstract
Integrin linked kinase (ILK) is a 59 kDa protein that has been previously implicated in both collagen- and thrombin-induced platelet activation. Platelet stimulation results in the transient activation of ILK kinase activity and movement to the plasma membrane where signaling complexes are formed with beta 1 and beta 3 integrins. The change in ILK activity and association with beta 1 and beta 3 integrins that occurs upon stimulation suggests that this protein may be important for the co-ordination of platelet responses. We have successfully developed a conditional ILK knockout mouse model using the Cre-Lox system to enable the study of platelets deficient in this protein. ILK deficient mice appear healthy and have normal platelet levels by day 8 following induction of gene deletion. ILK deficient platelets display reduced aggregation and fibrinogen binding in response to agonists such as collagen and thrombin. This is accompanied by a secretion defect, although early collagen stimulated signaling such as PLCγ2 phosphorylation and calcium mobilization are unaffected. Analysis of blood from ILK deficient mice using an in vitro flow system showed reduced thrombus formation under arterial conditions. Furthermore, extended bleeding times were observed in these mice. The data presented here demonstrates that ILK has a role in platelet regulation and is important for functional haemostasis.
The Novel Glycolysis Inhibitor, 3-BrOP, Has Broad Activity Against Acute Lymphoblastic Leukemias and Potentiates Methotrexate and Rapamycin.
Authors: Akers Lauren J. , Franklin Anna R. , Huang Peng , Zweidler-McKay Patrick A. 
Published: October 2019

Abstract
Background: Rapidly growing cancer cells are often dependent on glycolysis for ATP production, known as the Warburg effect. The novel glycolysis inhibitor, 3-bromo-2-oxopropionate-1-propyl ester (3-BrOP), has shown efficacy in models of glioblastoma, colon carcinoma and lymphoma. Evidence suggests that acute lymphoblastic leukemias (ALL) are also dependent on glycolysis. We proposed that potent inhibition of glycolysis would induce growth arrest and cell death in ALL blasts. Furthermore, we hypothesized that anti-metabolic therapies (methotrexate and rapamycin) may synergize with the ATP depletion caused by inhibition of glycolysis.
Objective: To determine the effect of 3-BrOP on human precursor-B and T cell leukemias, and to evaluate combination therapy with methotrexate (MTX) and/or rapamycin.
Design/Methods: A panel of nine human pre-B ALL and T-ALL cell lines were treated with 3-BrOP to determine IC50 via cell counts, cell cycle and MTT analysis under conditions of hypoxia and normoxia. Subsequently combinations with methotrexate and rapamycin were evaluated.
Results: All pre-B ALL and T-ALL cell lines demonstrate growth arrest, decreased MTT metabolism and significant apoptosis when treated with 3-BrOP alone (IC50 10–30mcM) in both normoxia and hypoxia (0.5% 02). Indeed 40mcM 3-BrOP induced >90% death in all cell lines tested. Both pre-B ALL and T-ALL cell lines undergo enhanced apoptosis when 3-BrOP is combined with modest doses of methotrexate (7.5nM) or rapamycin (50nM), as compared to these agents alone. Specifically, at 72 hours exposure T-ALL MTT proliferation decreased from 20% with 3-BrOP alone to 1% with 3-BrOP + rapamycin (p=0.037). Furthermore, sub-G0 content increased from 44% with 3-BrOP alone to 75% with 3-BrOP + rapamycin (p=0.032). Similarly, at 48 hours of exposure, pre-B ALL cells had an increased sub-G0 content from 36% with 3-BrOP alone to 47% with 3-BrOP + rapamycin (p=0.025).
Conclusions: Inhibition of glycolysis via 3-BrOP induces growth arrest and apoptosis across a panel of human pre-B and T cell ALL cell lines indicating a generalized dependence on glycolysis. 3-BrOP is equally effective in both hypoxia and normoxia. When combined with methotrexate and/or rapamycin, 3-BrOP potentiates their anti-leukemia effects. These combinations offer promising therapeutic approaches in ALL and support further pre-clinical evaluation.
Figure Figure
Potential Role of Dual NF-κB and Oxidative Stress Pathway Inhibitor, OT-304 as a Novel Drug Resistance-Reversing Agent.
Authors: Mousa Shaker A. , Patil Ghanshyam , Rebbaa Abdelhadi 
Published: October 2019

Abstract
The development of resistance to chemotherapy represents an adaptive biological response by tumor cells that leads to treatment failure and patient relapse. During the course of their evolution (intrinsic resistance) or in response to chemotherapy (acquired resistance), tumor cells may undergo genetic alterations to possess a drug resistant phenotype. Dysregulation of membrane transport proteins and cellular enzymes, as well as altered susceptibility to commit to apoptosis are among the mechanisms that contribute to the genesis of acquired drug resistance. Recently, the development of approaches to prevent and/or to reverse this phenomenon has attracted special interest and a number of drug candidates have been identified. Despite strong effects observed for these candidates in vitro, however, most of them fail in vivo. In the present study, we have identified a novel small molecule inhibitor of dual NF-κB and oxidative stress pathways, OT-304, as a potential candidate to reverse drug resistance. Initial investigations indicate that this compound effectively inhibits proliferation of doxorubicin-sensitive and doxorubicin-resistant cells to the same extent, suggesting that it is capable of bypassing the development of drug resistance. Additional experiments reveal that OT-304 enhances cancer cell sensitivity to doxorubicin and to etoposide, particularly in cells characterized by the over-expression of the drug transporter P-glycoprotein. These findings suggest that either the expression/and or the function of P-glycoprotein could be affected by OT-304. In vivo studies using tumor xenografts in nude mice showed that OT-304 is also capable of preventing the growth of drug resistant cancer cells. This later finding further confirms the role of OT-304 as a drug resistance-reversing agent and warrants further pre-clinical and clinical investigation to determine its efficacy in treating aggressive tumors.
Simple, Gel-Based Determination of Nucleophosmin Mutations in Acute Myeloid Leukemias.
Authors: Ahrens Kim P. 
Published: October 2019

Abstract
Mutations in the Nucleophosmin, member 1 (NPM1) gene, have been shown to be significant for prognosis and treatment of cytogenetically normal (CN) patients with acute myeloid leukemia (AML). To increase sensitivity for minimal residual disease (MRD) detection, especially in AMLs without a distinct phenotype or genetic marker, some assays have specifically targeted the individual mutation, which has led to an increased complexity of these tests. Other assays use expensive equipment such as capillary electrophoresis to differentiate between the usual 4bp size difference in the mutated gene and the wildtype(wt) gene. We developed a simple PCR-based procedure that amplifies the exon 12 region of the NPM1 gene and produces a small product (156bp for the wt and 160 for the mutant genes) using RNA that can be separated on a 4% metaphor agarose gel. This approach is clinically relevant since it can easily distinguish greater than 95% of the mutations. Eight of 24 AMLs of various types were positive for the mutation using our technique and those that were cloned and sequenced yielded the most common insertion TCTG, confirming its accuracy.
A Novel Approach for Analyzing Gene Expression Profiles Defines Two Distinct Subgroups of t(4;11) Positive Infant Acute Lymphoblastic Leukemia Patients.
Authors: Stam Ronald W. , Bohlander Stefan , Kirschner-Schwabe Renate , Arentsen-Peters Susan , Pieters Rob , Dingermann Theo , Marschalek Rolf 
Published: October 2019

Abstract
Gene expression profiles (GEPs) are becoming increasingly more important for diagnostic procedures, allowing clinical predictions including treatment response and outcome for various types of cancer. However, established gene signatures not always appear helpful in understanding underlying disease mechanisms. Therefore, we explored an alternative approach for analyzing GEPs of a group of t(4;11) positive infant acute lymphoblastic leukemia patients (n=15), a highly aggressive type of MLL rearranged leukemia. We developed a method that uses a relational database program in combination with a normalization approach and specific discriminators. For normalization, every GEP of a given t(4;11) positive ALL sample was compared with 3 GEPs of normal bone marrow aspirates derived from healthy donors. Using the GeneChip Analysis Suite 5.0 program for single comparison analysis, the resulting gene lists were then compared with each other and only consistently identified genes (present on all three gene lists) were used for further analysis. This noise reduction decreased the amount of potentially deregulated target genes to about 30–40%. The remaining gene lists represented highly significant target genes that were then incorporated into a relational database program using specific discriminators. These discriminators were:
upregulation of HOXA9 and HOXA10, presence/absence of the AF4-MLL fusion transcripts in addition to the MLL-AF4 fusion, and the localization of the genomic breakpoint within the MLL gene.
This pilot study led to promising results, surprisingly classifying individual t(4;11) positive ALL patients into two distinct subgroups. Both subgroups share about 80 target genes, but also display particular sets of subgroup specific target genes. Importantly, these identified target genes can directly be linked to biological properties of t(4;11) positive leukemia cells, and therefore, allow important novel insights into this aggressive type of leukemia in infants.
Expression of the Human PIM-2 (hPIM-2) Gene in Patients with Acute Myeloblastic Leukaemia (AML) and Acute Lymphoblastic Leukaemia (ALL).
Authors: Kapelko-Slowik Katarzyna L. , Urbaniak-Kujda Donata M. , Jazwiec Bozena , Dybko Jaroslaw , Wolowiec Dariusz , Slowik Miroslaw , Frydecka Irena , Kuliczkowski Kazimierz 
Published: October 2019

Abstract
Background. A human PIM-2 is proto-oncogene that encodes serine/threonine kinase which interacts with various signalling molecules. hPIM-2 is highly expressed in haematopoietic tissues and in leukaemic and lymphoma cell lines which is consistent with a role during haematopoiesis and during oncogenic transformation. The aim. The aim of the study was to investigate whether the human PIM-2 is altered in acute myeloblastic leukaemia (AML) and acute lymphoblastic leukaemia (ALL). Patients. Thirty-seven patients were included: 22 with AML and 15 with ALL (10 with T and 5 with B subtype), aged 21–70 (x=41). Twenty-two patients reached complete remission (CR). Bone marrow samples were collected at the time of diagnosis. Control samples were obtained from 8 healthy donors.
Methods. We analyzed hPIM-2 expression by quantitative RT-PCR analysis. RNA was isolated from 3mlns mononuclear cells by using standard methods (TriPure Reagent, Roche Diagnostics). cDNA was synthesized by AMV reverse transcriptase (Finnzyme). Sequence for primers was forward, 5′-CTTTCCTTCCAATACCCCA-3′ and reverse, 5′-CCATCTTCCATTCCTTCCC-3′. As a n internal control c-ABL was used with primers c-ABL: 5′-CCCAACCTTTTCGTTGCACTGT-3′ and 5′-TGA CTGGCGTGATGTAGTTGCTT-3′. The intensity of each band was compared densitometrically.
Results. Expression of the hPIM-2 in all leukemic patients (n=37) and both subgroups: AML (n=22) and ALL (n=15) was significantly higher than in normals (p=0,0001; p=0,0009; p=0.0002, respectively). Patients who reached CR expressed hPIM-2 significantly lower than patients with primary resistance to chemotherapy (with no CR) (p= 0,01). Moreover we have found the correlation between hPIM-2 expression and patients age (r=0,45, p=0,005) and WBC in peripheral blood (r=0,34, p=0,04). Summary. PIM-2 expression is increased in patients with AML and ALL Our data indicate that expression of hPIM-2 could be considered as an additional prognostic marker in patients with AML and ALL.