bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-12 Producing Capacity of Cultivated Dendritic Cells of Aplastic Anemia Patients during Disease Manifestation and Immunosuppressive Treatment.
Authors: Archuadze Shorena , Mikhailova Elena , Parovichnikova Elena , Kulikov Sergei , Savchenko Valeri 
Published: October 2019

Abstract
Dendritic cells (DCs) play principal role in the induction of antigen-specific T-cell immune response and in the development of antigen-specific self-tolerance. Moreover, they promote either Th1 or Th2 polarization of naive lymphocytes by production of immunoregulatory cytokines (IL-12 and IL-10). AA is characterized by increased Th1/Th2 ratio and by Th1 mediated antigen-specific suppression of hematopoiesis. DCs might be responsible for the activation of T-clone in AA patients. However, functional characteristics such as IL-12 production by DC in AA patients have not been studies yet. Therefore, IL-12 secretion capacity of DCs generated from peripheral mononuclear cells (PMNC) of 30 AA patients before and during the immunosuppressive therapy (IST) was compared to that of DCs of 7 donors. DC were derived from PMNC in the presence of GM-CSF and IL-4 for 5–7 days and further stimulated by exposition to 3T3-CD40L fibroblasts for 48 hours. Supernatants of DC cultures were subjected to ELISA for evaluation of IL-12 production. 70,8% of AA patients exhibited significantly (p=0,02) increased baseline level of IL-12 production by DCs compared to that of donors. Programmed IST resulted in the diminution of IL-12 production level in 67% of AA patients. Moreover, significant decrease in IL-12 production level correlated (Rs=0,79) with the achievement of independence from transfusions. Patients with high baseline level of IL-12 production by DCs required continuous (median - 24 months, p<0,05, see pict. 1) IST including repeated courses of antithymocyte globulin and cyclosporin-A. These data suggest that increase in IL-12 production by DCs of AA patients might contribute to cytotoxic T-clone expansion and consequently, to AA development. Patients that showed high initial levels of IL-12 secretion by DCs comprised the worst prognostic group.
Figure Figure
Effects of Sodium Valproate on Proliferation and Apoptosis of Human Myelodysplastic Syndromes Cell Line MUTZ-1.
Authors: Chen Bao-An , Zhao Hui-Hui , Xia Guo-Hua , Shao Ze-Ye , Gao Feng , Chen Jian , Sun Yun-Yu , Wang Jun , Zhao Gang , Ding Jia-Hua , Chen Ning-Na , Pei Xiao-Ping 
Published: October 2019

OBJECTIVE: Myelodysplastic syndromes (MDS) are a heterogeneous group with the expansion of a malignant clone. No satisfactory treatment for MDS is available. Sodium valproate (VPA), can induce G0–G1 phase arrest and cell apoptosis, inhibit proliferation of tumor cells in vitro significantly. The effects and possible mechanism of VPA on MUTZ-1 cell line of MDS were studied in this experiment.
METHODS: Cell proliferation was determined by MTT assay. Apoptotic morphological features were observed under microscope and transmission electronmicroscope. Cell apoptosis and cell cycle were analyzed by flow cytometry (FCM). The expression of p21WAF1(cyclin-dependent kinase inbihitor) was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.
RESULTS: VPA could inhibit the proliferation of MUTZ-1 cells in dose-and time-dependent manners. The typical apoptotic morphological features appeared in cells treated with 4 mmol/L VPA for 72 hours. Condensation of cells and nuclear chromatin, disintegration of nuclear chromatin, and apoptotic body could be observed under microscope. Aggregation and margination of apoptotic nuclear chromatin, cytoplasm condensation, and irregular chromatin masses could be observed under transmission electronmicroscope. The percentage of apoptotic cells which were treated with 1,2 and 4 mmol/L VPA for 72 hours increased from 1.39% to 2.18%, 16.03%, 22.02%, and cell cycle arrest at G0–G1 phase could be caused by VPA as shown by FCM. The expression level of p21WAF1 mRNA and p21WAF1 protein were up-regulated in a dose dependent manner in MUTZ-1 treated with VPA for 72 hours (P<0.05).
CONCLUSION: VPA induced a G0–G1 cell cycle arrest and have an effect on proliferation and apoptosis of MUTZ-1 cells in vitro. The mechanism may be associated with the up-regulation of p21WAF1.
Ectopic Expression of Mer on Jurkat Cells and Its Anti-Apoptosis Effect.
Authors: Fan Lei , Xie Liqian , Shen Fei , Dai Lan , Li Jianyong , Ruan Changgeng 
Published: October 2019

Abstract
BACKGROUND & OBJECTIVE: Mer is one member of Axl receptor tyrosine kinase family; its ligand Gas6 can bind Mer, then stimulate the tyrosine kinase activity and downstream cell signal pathway of Mer, and take part in cell inflammation, apoptosis and thrombosis. There are more and more reports on Mer function, while few on its association with malignant diseases. This study was to detect the expression of Mer on T-cell leukemia cell line--Jurkat, and investigate its anti-apoptosis function and the mechanism.
METHODS: Flow cytometry (FCM) was used to detect Mer expression on normal T cells and Jurkat cells. RNA interference (RNAi) was used to block the expression of Mer on Jurkat cells. The effect of Mer on the proliferation of Jurkat cells was assessed by MTT assay, and its effect on serum starvation-induced apoptosis was evaluated by FCM with Annexin V/PI double staining. The expression of apoptosis-associated genes Bcl-2 and Caspase-3 in Jurkat cells was detected by real-time polymerase chain reaction (PCR) after Mer blocking.
RUSELTS: Mer was not expressed in normal T cells both in peripheral blood and bone marrow, but highly expressed in Jurkat cells with a positive rate of 51.1%±5.2%. The inhibition rate of Mer expression in Jurkat cells by RNAi was 86.0%±10.3%. After 48-hour serum starvation, the apoptosis rate was 15.3%±4.3% in Mer-blocking Jurkat cells, and only 1.5%±1.0% in control Jurkat cells(P<0.01). There was no significant difference in Jurkat cell proliferation rate between these 2 groups. After Mer blocking, Bcl-2 expression was decreased by 42.7%±8.6% of control(P<0.01), Caspase-3 expression showed no significant change.
CONCLUSION: Mer is highly expressed in Jurkat cells, and could inhibit cell apoptosis via Bcl-2 signaling pathway.
A Novel Histone Deacetylase Inhibitor MCT-3 Augments Inhibition of Leukemia Cell Growth and Suppresses Induction of MMP-9 Expression by Azacitidine in HL-60 Cells.
Authors: Liu Hongbin , Mayes Penelope A. , Perlmutter Patrick , Mckendrick Joseph J. , Dear Anthony E. 
Published: October 2019

Abstract
Azacitidine (AZA), a demethylating agent, has recently been demonstrated to have efficacy in the treatment of myelodysplasia and acute myeloid leukemia. A potential concern when considering the use of this agent is a recent report demonstrating AZA-mediated re-activation of matrix metalloproteinase-9 (MMP-9) expression facilitating the invasive metastatic phenotype (Sato NM et al J Natl Cancer Inst. 2003 Feb 19;95(4):327–30). Histone deacetylase inhibitors (HDACi) are a recently identified class of agents with considerable in vitro and in vivo activity and early phase clinical efficacy in the treatment of haematological malignancies (Bolden JE Nat Rev Drug Discov. 2006 Sep;5(9):769–84). Pre-clinical in vitro studies suggest that addition of HDACi to 5-aza-2′-deoxycytidine, a derivative of AZA, enhances the efficacy of this agent (Yang H et al Leuk Res. 2005 Jul; 29(7):739–48) whilst early phase clinical trials identify therapeutic activity using a combination of demethylating agents and HDACi (Garcia-Manero G, Blood. 2006 Nov 15;108(10):3271–9). Our current study aimed to determine the in vitro activity and molecular mechanisms of action of the novel HDACi MCT-3, a derivative of Oxamflatin, a hydroxamate analogue, (Dear AE, et al, Org Biomol Chem, 2006, 4, 3778–3784) in the HL-60 cell line alone and combination with AZA. AZA (1.0 microM) and MCT-3 (2.5 microM) alone inhibited HL-60 cell growth over 24hrs by 40%, 30% respectively. The combination of AZA with MCT-3 inhibited HL-60 cell growth up to 50%. Real-time PCR demonstrated that AZA and MCT-3 alone increased p15INK4b and Caspase 3 mRNA expression 2 fold. A Combination of AZA with MCT-3 increased p15INK4b and Caspase 3 mRNA expression up to 2.5 and increased p21WAF1/CIP1 and the orphan nuclear receptor Nur77 expression 2 fold. A combination of AZA and MCT-3 significantly attenuated AZA-induced MMP-9 mRNA expression and proteolytic activity. AZA and MCT-3 alone reduce HL-60 cell growth in vitro. Addition of MCT-3 to AZA increased inhibition of cell growth, suggesting that this HDACi may have the potential for additive activity with demethylating agents. AZA and MCT-3 have similar effects on expression of genes implicated in cell cycle arrest and apoptosis. Increased expression of p21WAF1/CIP1 and the orphan nuclear receptor Nur77 via inhibition of cell cycle progression and enhanced apoptosis may in part be responsible for the enhanced anti-leukaemia activity of the combination of AZA and MCT-3. Importantly MCT-3 is able to inhibit AZA-mediated induction of MMP-9 expression.
Methylation of CpG Islands Was Involved in the Down-Regulation of DAZAP2 in Multiple Meyloma Cells.
Authors: Hu Wei-Xin , Ren Wei , He Le-Qi , Sheng Rong , Shi Yi-Wu 
Published: October 2019

Abstract
Our previous studies showed that DAZAP2 (deleted in azoospermia associated protein 2) was the most profoundly downregulated gene in bone marrow mononuclear cells from multiple meyloma (MM) patients. The results implicated a role for DAZAP2 as a potential tumor suppressor involved in the origin and development of MM. To further understand the molecular mechanism underlying this deregulation, we analyzed epigenetic changes (i.e., DNA methylation) associated with DAZAP2 in MM, given that aberrant promoter methylation contributes to tumorigenesis through inducing transcriptional suppression and tumor suppressor inactivation. Bioinformatics analysis showed that there are three CpG islands in the DAZAP2 promoter region. The fragments of DAZAP2 promoter region with different length were separately PCR-amplified and inserted into the luciferase reporter, pGL2-Basic vector. The reporter constructs were transfected into COS-7 cells and their luciferase activities measured. The results showed that the fragment containing the third CpG island was the most active among the constructs tested. When this fragment was treated with M. Sss I methylase, its luciferase activity was almost lost. On the other hand, after 5-aza-2′-deoxycytidine treatment, the expression of DAZAP2 was restored in KM3 cells, an MM cell line associated with DAZAP2 downregulation. To extend these observations, bone marrow samples from MM patients and normal controls were collected and genomic DNA, RNA and proteins were extracted from each sample for RT-PCR, MSP (methylation-specific PCR) and Western blot analyses. The results were in general agreement with that reported previously, demonstrating that the protein and mRNA of DAZAP2, while detectable in 100% normal controls, were undetectable in 62.5% MM patients. The MSP data further corroborated with RT-PCR and Western blot analysis (94%), which is characterized by a negative correlation between promoter methylation and downregulation of DAZAP2 in multiple myeloma. These results indicate that promoter hypermethylation may play an important role in downregulation of DAZAP2 gene expression in multiple myeloma.
Molecular Mechanisms of NF-kappaB Activation by the HTLV-I Oncoprotein Tax: Role of Post Translational Protein Modification.
Authors: Kfoury Youmna , Nasr Rihab , Favre-Bonvin Arnaud , El-Sabban Marwan , Renault Noemie , Giron Marie-Louise , Hajj Hiba El , Chiari Estelle , Mikati Abdul Ghani , Hermine Olivier , Saib Ali , de Thé Hugues , Pique Claudine , Bazarbachi Ali 
Published: October 2019

Abstract
Adult T-cell leukemia/lymphoma (ATL) is an aggressive malignancy of mature activated T-cells caused by human T cell lymphotropic virus type I (HTLV-I). The Tax viral oncoprotein plays a crucial role in the proliferation and transformation of HTLV-I infected T lymphocytes through various mechanisms, chiefly through activation of the NF-kappaB pathway. We found that ubiquitin binding to Tax C-terminal lysines in the cytoplasm is critical for Tax binding to the IkappaB kinase (IKK) complex and subsequent nuclear translocation of NF-kappaB. Conversely, we demonstrate that the same Tax lysines are modified by SUMO binding in the nucleus, an event required for the formation of Tax nuclear bodies and full NF-kappaB transcriptional activation. We further show that ubiquitylated Tax is not associated with active cytosolic IKK subunits, but binds endogenous IKK subunits and targets them to the centrosome. Indeed, differential ubiquitylation of Tax results in different outcomes: K63-ubiquitylated Tax colocalizes at the centrosome with IKK, while K48-ubiquitylated Tax is degraded by the proteasome. Altogether, these results support a model in which K63-ubiquitylated Tax activates IKK in a centrosome-associated signalosome, leading to the production of Tax-free active cytoplasmic IKK. Thus, ubiquitylation and sumoylation of the same lysine residues of Tax regulate two essential steps controlling NF-kappaB activation, demonstrating how these posttranslational modifications can cooperate to promote Tax-induced transformation. These observations highlight an unsuspected cellular and biochemical complexity in Tax-induced NF-kappaB activation and represents ideal targets for the development of new drugs for ATL therapy.
Gadd45a Behaves as a Tumor Suppressor and Gadd45b Behaves as an Oncogene in myc-Mediated Block in Myeloid Differentiation.
Authors: Mohamed-Hadley Alisha , Gibbs John D. , Liebermann Dan A. , Hoffman Barbara 
Published: October 2019

Abstract
We have observed that deregulated c-Myc expression in M1 myeloid leukemic cells blocks IL-6-induced differentiation at an intermediate stage, the cells fail to exit the cell cycle, proliferating indefinitely, with a portion of the cells always undergoing apoptotic cell death by prematurely recruiting the CD95/Fas apoptotic pathway. Similar results were also observed in normal myeloid cells from bone marrow (Amanullah et al. Oncogene19:2967–77, 2000; Amanullah et al. Oncogene21:1600–1610, 2002). Although there is evidence that c-myc represses the gadd45a promoter, this laboratory has observed elevated gadd45a expression in M1myc cells that are blocked during differentiation. In addition, microarray analysis indicated that gadd45b is a c-myc target gene. Because of these observations and the availability of gadd45a null and gadd45b null mice, the effect of deregulated myc expression in Hoxb8 immortalized macrophage progenitors was ascertained. It was observed that deregulated c-myc expression in gadd45a null cells blocked differentiation at the blast stage, compared to a block at the intermediate stage for wild type progenitors. In contrast, deregulated c-myc expression in gadd45b null progenitors resulted in rapid cell death. These observations suggest that in the context of myc mediated myeloid leukemias Gadd45a behaves as a tumor suppressor and Gadd45b behaves as an oncogene.
Direct and Natural Killer Cell-Mediated Anti-Tumor Effects of Low-Dose Bortezomib in Hepatocellular Carcinoma.
Authors: Krusch Matthias , Armeanu Sorin , Baltz Katrin M. , Lauer Ulrich M. , Steinle Alexander , Kanz Lothar , Bitzer Michael , Salih Helmut R. 
Published: October 2019

Abstract
Background & Aims: Hepatocellular carcinoma (HCC) displays a particular resistance to conventional cytostatic agents. Therefore, alternative treatment strategies focus on novel substances exhibiting anti-neoplastic and/or immunomodulatory activity enhancing for example Natural Killer (NK) cell anti-tumor reactivity. However, the tumor-associated ligands engaging activating NK cell receptors are largely unknown. Exceptions are the NKG2D ligands (NKG2DL) of the MIC and ULBP protein families, which potently stimulate NK cell responses. We studied the consequences of proteasome inhibition with regard to direct and NK cell-mediated effects against HCC.
Methods: Primary human hepatocytes (PHH) from different donors, hepatoma cell lines and NK cells were exposed to Bortezomib. Growth and viability of hepatoma cells and PHH as well as immunomodulatory effects including alterations of NKG2DL expression on hepatoma cells, specific induction of NK cell cytotoxicity and IFN-γ production were investigated.
Results: Bortezomib treatment inhibited hepatoma cell growth with IC50 values between 2.4 and 7.7 nanomol/liter. These low doses increased MICA/B mRNA levels and total and cell surface protein expression in hepatoma cells, which stimulated cytotoxicity and IFN-γ production of cocultured NK cells. Importantly, while IFN-γ production of NK cells was concentration-dependently reduced, low-dose Bortezomib neither induced NKG2DL expression or cell death in PHH nor altered NK cell cytotoxicity.
Conclusions: Low-dose Bortezomib mediates a specific dual anti-tumor effect in HCC by inhibiting tumor cell proliferation and by priming hepatoma cells for NK cell anti-tumor reactivity. Our data suggest the clinical evaluation of Bortezomib treatment in HCC, especially in combination with immunotherapeutic approaches like adoptive NK cell transfer.
Imatinib Mesylate Induced Reversal of Leukemic Gene Phenotype in HL60 Cells Coincides with Stimulation of Oxidative Metabolism.
Authors: Pfeilstocker Michael , Wihlidal Peter , Varga Franz , Pittermann Elisabeth , Karlic Heidrun 
Published: October 2019

Abstract
Besides blockade of tyrosine kinases such as c-kit, Imatinib mesylate (IM) regulates glucose flux through downregulation of GLUT-1 transporters in human leukemia cells. This mechanism has the potential to induce regression of type 2 diabetes and hyperlipidemia as observed in patients with chronic myeloid leukemia or hypereosinophilic syndrome. In addition, there is a stimulatory effect of IM on differentiation of human mesenchymal stem cells. Its synergism with retinoic acid or low dose Ara-C is applied in treatment of acute myeloid leukemia (AML). Thus, the AML-derived c-kit positive cell line HL60 was chosen for studying the effect of IM on expression of genes associated with differentiation and metabolism. We analysed the possible feedback on transcription factors (AML1 and AML3) and consequences regarding differentiation and metabolism - associated genes. Quantitative reverse transcriptase PCR analyses revealed that IM treatment of HL60 cells downregulates mRNA synthesis of AML1 and AML3 by 70% without affecting transcription of the c-abl tyrosine kinase. IM reduces expression of CD34 mRNA from 20% to 6% of the housekeeping gene G6PD. The appearance of differentiated cells was accompanied by a remarkable stimulation of mRNAs from CD11b and CD14 (monocyte markers) reaching 4-fold higher expression levels relative to G6PD. This was associated with an increased proportion of osteocalcin (OCN), which has recently been shown to enhance mitochondrial activity. A 2-fold stimulation of a fat-metabolism associated mitochondrial palmitoyltransferase (CPT1B) and 10-fold stimulation of microsomal carnitine palmitoyltransferase and the carnitine transporter OCTN2 supports previous data indicating an IM-associated stimulation of oxidative metabolism resulting in a regression of type-2 diabetes and hyperlipidemia. Our current investigations show that IM-associated attenuation of cell proliferation inhibited transcription factors AML1 and AML3 and triggered differentiation in the leukemic cell line HL60, as reflected by altered mRNA expression of surface marker genes. The IM - induced stimulation of lipid metabolism in HL60 confirms previous data indicating a reversal of the Warburg effect in K562 cells without cytocidal activity. This indicates a similar mechanism as known for other drugs and strategies targeting glucose or fat metabolism, which are discussed in the context of cancer prevention and treatment.
A Novel Chemo Sensitivity Index for Melphalan Based on Gene Expression Profiling (GEP).
Authors: Dahlgaard Jesper O. , Dybkaer Karen , Johnsen Hans Erik , Knudsen Steen 
Published: October 2019

Abstract
Multiple Myeloma (MM) is a lethal hematological malignancy with an incidence of 40–60 per 1.000.000 per year. Over the last decade, the most significant therapeutic improvement has been the introduction of high-dose Melphalan therapy in combination with autologous stem cell support. This strategy has led to an increase in survival from a median of approximately 3 years till now >6 years also in Denmark (unpublished data from Nordic Myeloma Study Group, NMSG #7/98). Despite this important increase in survival, substantial variability still persists among patients in response to high-dose Melphalan. Clinical and genetic heterogeneity is a characteristic feature of MM, and new methodological approaches, such as global gene expression profiling (GEP), are valuable tools in order to link clinical endpoints, such as response to treatment and survival, with the molecular and genetic heterogeneity of the disease (Shaughnessy, J. et al., Blood98, 217–223, 2001). Here we present a bioinformatic analysis of the clinical impact of a novel chemo sensitivity index based on GEP of 1) a panel of sixty cell lines (NCI60) that subsequently have been exposed to a range of anti-cancer agents, including Melphalan, and, 2) a retrospective validation of our method from lung, brain and lymphoma patients. Figure 1, presents a retrospective leave-one-out cross-validation of the prediction whether a patient will respond to the treatment given. Lung- and brain cancer patients were treated with Cisplatin, and the predicted sensitivity or resistance to this drug was used to group patients (hazard ratio 2.9, 95% CI 0.7–11). Lymphoma patients were treated with Vincristine and Adriamycine, and the patients were grouped according to their predicted sensitivity or resistance to these drugs. Patient and raw DNA chip data were obtained from: Beer et al, Nature Medicine 8(8):816–824; Pomeroy et al, Nature 415(6870):436–442; Shipp et al, Nature Medicine 8(1):68–74. For each patient, the method may also allow for a quantitative prediction of drug sensitivity towards a range of drugs including Melphalan. Figure 2 shows the drug sensitivity profile for two selected Multiple Myeloma patients (patient #2 and #9 from Agnelli L et al. Haematologica. 2007 Jan; 92(1):56–65). We are currently collecting chip data from a cohort of 70 newly diagnosed biobanked bone marrow samples from MM patients treated with high-dose Melphalan in NMSG trial #9/99 in order to further confirm these results in MM. These data are subsequently being validated using an independent population of bone marrow samples from MM patients also treated with high-dose Melphalan. We demonstrate that GEP can provide new information concerning resistance towards Melphalan and a range of anti-cancer drugs, separately or in combination (multi drug regimen), which may be clinically useful, and serve as a guide for the future development of individualized treatment strategies.
Figure Figure