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European Journal of Pharmacology Oct 2011Zinc (Zn) has been known to inhibit osteoclastic bone resorption and stimulate osteoblastic bone formation. However, the mechanisms responsible for these effects have...
Zinc (Zn) has been known to inhibit osteoclastic bone resorption and stimulate osteoblastic bone formation. However, the mechanisms responsible for these effects have not been well characterized in vivo. Here, the effects of a dietary administration of Zn on osteoclastogenesis and osteoblastogenesis were investigated in Zn-adequate rats. The administration of Zn decreased the activities of bone tartrate-resistant acid phosphatase (TRAP) and cathepsin K, without affecting the serum osteocalcin level. Histological analysis showed a decrease in the number of osteoclasts with a normal number of osteoblasts in the metaphysis of the proximal tibia. The mRNA levels of receptor for activation of NF-κB (RANK), c-fos, c-jun, TRAP and cathepsin K were significantly decreased, although those of RANK ligand, macrophage colony-stimulating factor and c-fms were unaltered. The gene expression of bone morphogenic protein-2, Runx2, Dlx5, osterix, alkaline phosphatase, osteocalcin and collagen was not affected. The level of the RANK protein decreased, while the levels of the Runx2 and β-catenin proteins were unchanged. Further, the osteoclastic differentiation of precursor cells in vitro was suppressed. The suppressed osteoclastogenesis was associated with decreased levels of reactive oxygen species, extracellular signal-regulated kinase (ERK) activation and RANK expression. A lower lipid peroxide level and a higher glutathione level were also observed. These results suggested that Zn-administration did not affect osteoblastogenesis but decreased osteoclastogenesis by inhibiting RANK expression through suppression of the production of reactive oxygen species and ERK activation in Zn-adequate rats.
Topics: Animals; Biomarkers; Cell Count; Cell Differentiation; Core Binding Factor Alpha 1 Subunit; Down-Regulation; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Female; Macrophage Colony-Stimulating Factor; Monocyte-Macrophage Precursor Cells; Osteoblasts; Osteoclasts; Rats; Rats, Wistar; Reactive Oxygen Species; Receptor Activator of Nuclear Factor-kappa B; Receptor, Macrophage Colony-Stimulating Factor; Signal Transduction; Tibia; Zinc; beta Catenin
PubMed: 21806983
DOI: 10.1016/j.ejphar.2011.07.003 -
Oncotarget Sep 2015Recent discoveries have led to the testing of novel targeted therapies for the treatment of acute myeloid leukemia (AML). To better inform the results of clinical...
Recent discoveries have led to the testing of novel targeted therapies for the treatment of acute myeloid leukemia (AML). To better inform the results of clinical trials, there is a need to identify and systematically assess biomarkers of response and pharmacodynamic markers of successful target engagement. Spleen tyrosine kinase (SYK) is a candidate therapeutic target in AML. Small-molecule inhibitors of SYK induce AML differentiation and impair leukemia progression in preclinical studies. However, tools to predict response to SYK inhibition and to routinely evaluate SYK activation in primary patient samples have been lacking. In this study we quantified phosphorylated SYK (P-SYK) in AML cell lines and establish that increasing levels of baseline P-SYK are correlated with an increasing sensitivity to small-molecule inhibitors targeting SYK. In addition, we found that pharmacological inhibition of SYK activity extinguishes P-SYK expression as detected by an immunohistochemical (IHC) test. Quantitative analysis of P-SYK expression by the IHC test in a series of 70 primary bone marrow biopsy specimens revealed a spectrum of P-SYK expression across AML cases and that high P-SYK expression is associated with unfavourable outcome independent of age, cytogenetics, and white blood cell count. This study thus establishes P-SYK as a critical biomarker in AML that identifies tumors sensitive to SYK inhibition, identifies an at-risk patient population, and allows for the monitoring of target inhibition during treatment.
Topics: Antineoplastic Agents; Biomarkers, Tumor; Bone Marrow; Bone Marrow Examination; Dose-Response Relationship, Drug; Enzyme Activation; HL-60 Cells; Humans; Immunohistochemistry; Intracellular Signaling Peptides and Proteins; Kaplan-Meier Estimate; Leukemia, Myeloid, Acute; Molecular Targeted Therapy; Phosphorylation; Predictive Value of Tests; Prognosis; Protein Kinase Inhibitors; Protein-Tyrosine Kinases; Risk Factors; Signal Transduction; Syk Kinase; U937 Cells; Up-Regulation
PubMed: 26315286
DOI: 10.18632/oncotarget.4669 -
Toxicon : Official Journal of the... Nov 2013BF-CT1, a 13 kDa protein isolated from Bungarus fasciatus snake venom through CM cellulose ion exchange chromatography at 0.02 M NaCl salt gradient showed cytotoxicity...
BF-CT1, a 13 kDa protein isolated from Bungarus fasciatus snake venom through CM cellulose ion exchange chromatography at 0.02 M NaCl salt gradient showed cytotoxicity in in vitro and in vivo experimental models. In in vivo Ehrlich ascites carcinoma (EAC) induced BALB/c mice model, BF-CT1 treatment reduced EAC cell count significantly through apoptotic cell death pathway as evidenced by FACS analysis, increased caspase 3, 9 activity and altered pro, antiapoptotic protein expression. BF-CT1 treatment caused cell shrinkage, chromatin condensation and induced apoptosis through increased caspase 3, caspase 9 activity, PARP cleavage and down regulation of heat shock proteins in U937 leukemic cell line. Cytosolic cytochrome C production was increased after BF-CT1 treatment upon U937 cell line. BF-CT1 treated U937 cell showed cell cycle arrest at sub G1 phase through cyclin D and CDK down regulation with up regulation of p15 and p16. It also down regulated PI3K/AKT pathway and MAPkinase pathway and promoted apoptosis and regulated cell proliferation in U937 cells. BF-CT1 prevented angiogenesis in in vitro U937 cell line through decreased VEGF and TGF-β1 production.
Topics: Angiogenesis Inducing Agents; Animals; Annexin A5; Apoptosis; Bungarus; Caspase 3; Caspase 8; Caspase 9; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Cyclin D; Cyclin-Dependent Kinases; Disease Models, Animal; Down-Regulation; Humans; Lethal Dose 50; Male; Mice; Mice, Inbred BALB C; Molecular Weight; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Reactive Oxygen Species; Reptilian Proteins; Signal Transduction; Snake Venoms; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Transforming Growth Factor beta1; U937 Cells; Up-Regulation; Vascular Endothelial Growth Factor A
PubMed: 23981271
DOI: 10.1016/j.toxicon.2013.08.052 -
Cancer Immunology, Immunotherapy : CII Mar 2012Severe immune suppression is frequent in late-stage tumor patients and promotes tumor immune evasion and subsequent tumor progression. Regulatory T cells (Treg) are... (Clinical Trial)
Clinical Trial
Severe immune suppression is frequent in late-stage tumor patients and promotes tumor immune evasion and subsequent tumor progression. Regulatory T cells (Treg) are major suppressors of anti-tumor immune responses. Therefore, targeting of Treg has become a key goal of anti-tumor therapy. Several preclinical and clinical observations suggest that Treg can be depleted by cyclophosphamide. Over a period of 3 months, we investigated the effect of metronomic low-dose cyclophosphamide on Treg numbers, suppressive capacity and proliferation on endogenous anti-tumor T-cell responses and on their correlation to clinical outcome in 12 patients with treatment-refractory metastasized breast cancer who received single-agent 50 mg cyclophosphamide p.o. daily. Cyclophosphamide treatment initially caused a significant reduction in circulating Treg by more than 40% (P = 0.002). However, Treg numbers completely recovered during the treatment due to increased proliferative activity and maintained their suppressive capacity. Treg depletion coincided with a strong increase in breast tumor-reactive T cells (P = 0.03) that remained at high levels during the whole period. Numbers of tumor-reactive T cells but not of Treg correlated with disease stabilization (P = 0.03) and overall survival (P = 0.027). We conclude that metronomic low-dose cyclophosphamide only transiently reduces Treg but induces stable tumor-specific T-cell responses, which correlate with improved clinical outcome in advanced-stage breast cancer patients.
Topics: Adult; Aged; Antineoplastic Agents, Alkylating; Breast Neoplasms; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Cyclophosphamide; Dose-Response Relationship, Drug; Female; Humans; Interferon-gamma; Lymphocyte Count; Middle Aged; Neoplasm Metastasis; Survival Analysis; T-Lymphocytes, Cytotoxic; T-Lymphocytes, Regulatory; Time Factors; Treatment Outcome; U937 Cells
PubMed: 21915801
DOI: 10.1007/s00262-011-1106-3 -
Gynecologic Oncology Aug 2015Tumor-associated macrophages are known to be associated with decreased survival of patients with endometrial cancer. Given the physiological link of circulating...
OBJECTIVE
Tumor-associated macrophages are known to be associated with decreased survival of patients with endometrial cancer. Given the physiological link of circulating monocytes as a progenitor of tumor-associated macrophages, monocyte counts were examined for tumor characteristics and survival in endometrial cancer.
METHODS
A retrospective study was conducted to examine consecutive patients with endometrial cancer with all histologic types who underwent hysterectomy-based surgical staging between 2003 and 2013 (n=541). Preoperative monocyte counts were correlated to patient demographics, pathological findings, complete blood count results, and survival outcomes.
RESULTS
Median monocyte counts were 0.5×10(9)/L. Monocyte counts significantly correlated with all other complete blood count components, with neutrophil counts having the most significant association (r=0.52, p<0.001). Elevated monocyte counts (defined as >0.7×10(9)/L) when compared to lower counts were significantly associated with an increased risk of >50% myometrial tumor invasion (29.2% versus 22.0%, odds ratio [OR] 1.59, 95% confidence interval [CI] 1.01-2.45, p=0.045), pelvic lymph node metastasis (39.0% versus 18.8%, OR 2.76, 95%CI 1.35-5.62, p=0.007), and advanced-stage (stage I through IV, 18.5%, 24.6%, 32.5%, and 41.5%, p=0.001). In survival analysis, elevated monocyte counts were associated with decreased disease-free survival (5-year rates, 71.0% versus 84.5%, p=0.001) and overall survival (77.2% versus 89.3%, p<0.001). In multivariate analysis, elevated monocyte counts remained an independent prognostic factor for decreased disease-free (hazard ratio [HR] 1.74, 95% CI 1.02-2.96, p=0.041) and overall (HR 2.63, 95% CI 1.37-5.05, p=0.004) survival.
CONCLUSIONS
Elevated monocyte counts were associated with aggressive tumor features and poor survival outcomes of patients with endometrial cancer.
Topics: Adult; Aged; Aged, 80 and over; Endometrial Neoplasms; Female; Humans; Macrophages; Middle Aged; Monocyte-Macrophage Precursor Cells; Monocytes; Neoplasm Staging; Prognosis; Retrospective Studies; Survival Rate; Young Adult
PubMed: 26013698
DOI: 10.1016/j.ygyno.2015.05.019 -
Bone 1994Age-associated osteopenia has been documented to occur in mice and, therefore, provides a model system whereby mechanisms of bone loss can be assessed in vivo and in...
Age-associated osteopenia has been documented to occur in mice and, therefore, provides a model system whereby mechanisms of bone loss can be assessed in vivo and in vitro. One such mechanism, that could explain the increased resorptive activity seen in some forms of osteopenia, is an age-associated increase in the osteoclast precursor pool and osteoclastogenic formation. To test this hypothesis, we studied the bone marrow composition of aged (24 months) mice to determine if increased numbers of monocyte/macrophage/osteoclast precursor cells (MMOPC) were present when compared to young (4-6 months) animals. Our data show a moderate increase of 20-30% more hematopoietic cells obtained from the long bones of the aged animals. However, both liquid and semi-solid culture techniques demonstrate an approximately 2-3.5-fold increase in the numbers of plastic adherent macrophages or mononuclear colonies in bone marrow derived from the aged mice when stimulated by interleukin-3 (IL-3), granulocyte-macrophage colony stimulating factor (GM-CSF) or macrophage colony stimulating factor (GM-CSF), indicating a preferential increase in MMOPCs. In addition, cells derived from the aged mice show higher levels of cytokine stimulated incorporation of [3H]-thymidine and [3H]-leucine, with increased protein synthesis seen up to 7 days after cytokine stimulation, suggesting that these cells also have an enhanced sensitivity to cytokines.(ABSTRACT TRUNCATED AT 250 WORDS)
Topics: Aging; Animals; Bone Marrow; Cell Count; Cells, Cultured; Mice; Mice, Inbred C57BL; Osteoclasts; Osteoporosis; Stem Cells
PubMed: 8024854
DOI: 10.1016/8756-3282(94)90893-1 -
Molecular Medicine Reports May 2016Neutrophil elastase (NE) is a neutrophil‑derived serine proteinase with specificity for a broad range of substrates. NE has been reported to be associated with the...
Neutrophil elastase (NE) is a neutrophil‑derived serine proteinase with specificity for a broad range of substrates. NE has been reported to be associated with the pathogenesis of several conditions, particularly that of pulmonary diseases. Previous studies have shown that NE can cleave the pro‑myelocyte ‑ retinoic acid receptor‑alpha chimeric protein and is important for the development of acute pro‑myelocytic leukemia. To further elucidate the role of NE in acute pro‑myelocytic leukemia, the present study successfully constructed a lentiviral vector containing the NE gene (LV5‑NE), which was transfected into NB4 acute pro‑myelocytic leukemia cells. The effects of NE overexpression in NB4 cells were detected using a Cell-Counting Kit‑8 assay, flow cytometry and western blot analysis. The results showed that NE significantly promoted the proliferation of NB4 cells, inhibited cell apoptosis and apoptotic signaling, and led the activation of Akt. In an additional experiment, a vector expressing small hairpin RNA targeting NE was constructed to assess the effects of NE knockdown in U937 cells. Western blot analysis revealed that apoptotic signaling was increased, while Akt activation was decreased following silencing of NE. The results of the present study may indicate that NE activates the phosphoinositide-3 kinase/Akt signaling pathway in leukemia cells to inhibit apoptosis and enhance cell proliferation, and may therefore represent a molecular target for the treatment of pro‑myelocytic leukemia.
Topics: Apoptosis; Cell Proliferation; Enzyme Activation; Humans; Leukemia; Leukocyte Elastase; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; U937 Cells
PubMed: 27035679
DOI: 10.3892/mmr.2016.5051 -
Indian Journal of Experimental Biology Sep 2016Hydantoin derivatives, including phenytoin (5,5-diphenylhydantoin), have recently gained attention as they possess a variety of important biochemical and pharmacological...
Hydantoin derivatives, including phenytoin (5,5-diphenylhydantoin), have recently gained attention as they possess a variety of important biochemical and pharmacological properties. Nevertheless, available information on anticancer activity of hydantoin derivatives is still scarce. Here, we evaluated possible antileukemic potential of four phenytoin analogs, namely: methyl 2-(2,4-dioxo-5,5-diphenylimidazolidin-3-yl)propanoate (1), methyl 2-(1-(3-bromopropyl)-2,4-dioxo-5,5-diphenylimidazolidin-3-yl)propanoate (2), 1-(3-bromopropyl)-3-methyl-5,5-diphenylimidazolidine-2,4-dione (3) and 1-(3-bromobutyl)-3-methyl-5,5-diphenylimidazolidine-2,4-dione (4). The experiments were performed on human acute histiocytic lymphoma U937 cells and human promyelocytic leukemia HL-60 cells. The present study was conducted using spectrophotometric 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay and the electronic Beckman-Coulter method. We observed temporary changes in the leukemia cell viability, volume and count. The effects of the four 5,5-diphenylhydantoin derivatives on U937 and HL-60 cells depended on the agent tested and its concentration, the time intervals after the compound application, and the leukemia cell line used. HL-60 cells were more sensitive than U937 cells to the action of the phenytoin analogs (1-4). The antileukemic activities of the three bromoalkyl diphenylhydantoin derivatives (2, 3, and 4) were stronger than that of the compound 1 [methyl 2-(2,4-dioxo-5,5-diphenylimidazolidin-3-yl) propanoate], with no bromoalkyl substituent. The structural modifications of 5,5-diphenylhydantoin are responsible for such varied antileukemic potential of its four derivatives.
Topics: Cell Survival; HL-60 Cells; Humans; Leukemia; Phenytoin; Structure-Activity Relationship; U937 Cells
PubMed: 28699720
DOI: No ID Found -
International Journal of Oncology Apr 2009Anthocyanins are a class of flavonoids, widely spread throughout the plant kingdom, that exhibit important anti-oxidant and anti-inflammatory actions as well as...
Anthocyanins are a class of flavonoids, widely spread throughout the plant kingdom, that exhibit important anti-oxidant and anti-inflammatory actions as well as chemotherapeutic effects. However, little is known concerning the molecular mechanisms by which these activities are exerted. In this study, we investigated the anthocyanins isolated from Vitis coignetiae Pulliat for their potential anti-proliferative and apoptotic effects on human leukemia U937 cells. It was found that these anthocyanins inhibit cell viability and induce apoptotic cell death of U937 cells in a dose-dependent manner, as measured by hemocytometer counts, by alteration in the mitochondrial membrane potential, by increases in sub-G1 populations and by DNA ladder formation. Apoptosis of U937 cells by anthocyanins was associated with modulation of expression of Bcl-2 and IAP family members. Consequently, anthocyanin treatment induced proteolytic activation of caspase-3, -8 and -9, and a concomitant degradation of poly(ADP-ribose) polymerase. However, anthocyanin-induced growth inhibition and apoptosis were significantly attenuated in Bcl-2 overexpressing U937 cells. Furthermore, z-DEVD-fmk, a caspase-3 specific inhibitor, blocked apoptosis and increased the survival of anthocyanin-treated U937 cells. Taken together, these results show that Bcl-2 and caspases are key regulators of apoptosis in response to anthocyanins in human leukemia U937 cells.
Topics: Anthocyanins; Apoptosis; Caspases; Cell Survival; DNA; Down-Regulation; Enzyme Activation; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Membrane Potentials; Mitochondrial Membranes; Proto-Oncogene Proteins c-bcl-2; U937 Cells; Vitis
PubMed: 19287965
DOI: 10.3892/ijo_00000234 -
Medical Science Monitor : International... Oct 2019BACKGROUND Acute myeloid leukemia (AML) is associated with a high relapse rate and poor prognosis. This study aimed to use weighted gene coexpression network analysis...
Weighted Gene Coexpression Network Analysis Identifies Cysteine-Rich Intestinal Protein 1 (CRIP1) as a Prognostic Gene Associated with Relapse in Patients with Acute Myeloid Leukemia.
BACKGROUND Acute myeloid leukemia (AML) is associated with a high relapse rate and poor prognosis. This study aimed to use weighted gene coexpression network analysis (WGCNA) of gene coexpression networks to identify candidate prognostic biomarker genes in patients with AML and to investigate the expression of these genes in the human U937 cell line in vitro. MATERIAL AND METHODS RNA-seq data were retrieved from the Cancer Genome Atlas (TCGA) and included bone marrow samples and survival data of patients with AML (N=151), patients who did not relapse after treatment (N=119), and patients with relapse (N=40). Differentially expressed genes were identified, WGCNA was used to detect functional modules, and survival analysis was performed. The Cell Counting Kit-8 (CCK-8) assay investigated the proliferation of U937 cells transfected with short hairpin RNAs (shRNAs), shCRIP1, shHIST1H1C, and shHIST1H1E. RNA-seq analysis identified gene expression following CRIP1 knockdown. RESULTS Eighty-two genes were associated with both relapse and prognosis in patients with AML. There were two prognosis-related gene modules in the coexpression network. In the coexpression network, the histone cluster 1 H1 family member gene, HIST1H1C had the maximum relapse fold change, HIST1H1E had the lowest survival p-value, and the cysteine-rich intestinal protein 1 (CRIP1) gene had the most edge numbers and was significantly associated with poor prognosis (P=0.0165786). RNA-seq data showed that there was a significant difference in gene expression after CRIP1 knockdown in U937 cells. CONCLUSIONS WGCNA of gene coexpression networks identified CRIP1 as a potential prognostic biomarker gene in patients with AML.
Topics: Carrier Proteins; Cell Proliferation; China; Computational Biology; Databases, Genetic; Female; Gene Expression Profiling; Gene Regulatory Networks; Humans; LIM Domain Proteins; Leukemia, Myeloid, Acute; Male; Prognosis; RNA, Long Noncoding; RNA, Small Interfering; Recurrence; U937 Cells
PubMed: 31577790
DOI: 10.12659/MSM.918092