acid-phosphatase and Neuroblastoma

Topics: 17-Hydroxycorticosteroids; 17-Ketosteroids; Acid Phosphatase; Adrenal Gland Neoplasms; Alkaline Phosphatase; Amino Acids; Amylases; Bone Neoplasms; Breast Neoplasms; Carcinoid Tumor; Catecholamines; Chorionic Gonadotropin; Clinical Enzyme Tests; Clinical Laboratory Techniques; Female; Glucose-6-Phosphate Isomerase; Humans; Hydroxyindoleacetic Acid; L-Lactate Dehydrogenase; Liver Neoplasms; Male; Neoplasms; Neoplasms, Nerve Tissue; Neuroblastoma; Nucleotidases; Pancreatic Neoplasms; Pheochromocytoma; Pregnancy; Prostatic Neoplasms; Trophoblastic Neoplasms; Vanilmandelic Acid

The progressive accumulation of misfolded and aggregated proteins in neurons is an accepted mechanism in aging. Overproduction of reactive oxygen species (ROS), referred to as oxidative stress, is currently believed to play a pivotal role in this process. Lipofuscin as a histological index of aging results from cross-links between oxidized proteins and lipids. Therefore, to attenuate lipofuscin formation, it would be logical to use exogenous natural or synthetic antioxidants. Yakuchinone B (1-[4'-hydroxy-3'-methoxyphenyl]-7-phenylhept-1-en-3-one) is a component of Alpinia oxyphylla seeds with established antioxidant activity.. To evaluate the neuroprotective roles of yakuchinone B (JC6) and its structural analogues (JC1-JC5), the free radical scavenging capabilities of yakuchinone B derivatives were studied in terms of cell viability, apoptosis, cells ROS content, catalase (CAT) and superoxide dismutase (SOD) activity and the intracellular lipofuscin content in SK-N-MC cells exposed to H2O2. The level of MDA (malondialdehyde), as an index of lipid peroxidation and acid phosphatase activity were also measured.. Our results indicated that derivatives especially JC4, JC5 and JC6 decreased the extent of apoptosis and ROS level, while they increased the activities of SOD and CAT in drug-pretreated cells as compared to H2O2-treated cells. A clear relationship between the structure and antioxidant activities of these compounds was established. In addition, JC4, JC5 and JC6 were capable of down-regulating the formation of MDA and lipofuscin.. Our results indicated that free radicals play significant roles in lipofuscin formation and cellular aging which can be attenuated by yakuchinone B derivatives.

Topics: Acid Phosphatase; Antioxidants; Apoptosis; Catalase; Cell Line, Tumor; Cell Survival; Chalcone; Diarylheptanoids; Free Radical Scavengers; Humans; Hydrogen Peroxide; Intracellular Space; Lipid Peroxidation; Lipofuscin; Neuroblastoma; Protective Agents; Superoxide Dismutase

Most newly synthesized proteins destined for the lysosome reach this location via a specific intracellular pathway. In the Golgi, a phosphotransferase specifically labels lysosomal proteins with mannose 6-phosphate (Man-6-P). This modification is recognized by receptors that target the lysosomal proteins to the lysosome where, in most cell types, the Man-6-P recognition marker is rapidly removed. Despite extensive characterization of this pathway, the enzyme responsible for the removal of the targeting modification has remained elusive. In this study, we have identified this activity. Preliminary investigations using a cell-based bioassay were used to follow a dephosphorylation activity that was associated with the lysosomal fraction. This activity was high in the liver, where endogenous lysosomal proteins are efficiently dephosphorylated, but present at a much lower level in the brain, where the modification persists. This observation, combined with an analysis of the expression of lysosomal proteins in different tissues, led us to identify acid phosphatase 5 (ACP5) as a candidate for the enzyme that removes Man-6-P. Expression of ACP5 in N1E-115 neuroblastoma cells, which do not efficiently dephosphorylate lysosomal proteins, significantly decreased the steady state levels of Man6-P glycoproteins. Analysis of ACP5-deficient mice revealed that levels of Man-6-P glycoproteins were highly elevated in tissues that normally express ACP5, and this resulted from a failure to dephosphorylate lysosomal proteins. These results indicate a central role for ACP5 in removal of the Man-6-P recognition marker and open up new avenues to investigate the importance of this process in cell biology and medicine.

Topics: Acid Phosphatase; Animals; Cell Line, Tumor; Glycoproteins; Humans; Isoenzymes; Mannosephosphates; Mice; Mice, Knockout; Neuroblastoma; Phosphorylation; Protein Processing, Post-Translational; Proteins; Tartrate-Resistant Acid Phosphatase

Neuroblastoma originates from neural crest cells and is the most common extracranial solid tumor in childhood. Bone metastasis in neuroblastoma is an unfavorable prognostic factor even with intensive therapy. In the present study, we screened four cell lines of human neuroblastoma (NB-1, NB-16, NB-19, and NH-6) for tumorigenicity and metastatic capacity in nude mice and found that NB-19 cells caused osteolytic lesions after s.c. injection into mice. To detect micrometastases in the host tissue, we performed two kinds of PCR-based metastasis assays: (a) genomic PCR assay using the primers for human genome-specific Alu sequence; and (b) reverse transcription-nested PCR assay that detects the expression of tyrosine hydroxylase, a marker specific for neuroblastoma. The results of these PCR assays revealed the colonization of human neuroblastoma cells in the bone marrow of the mice that had received the s.c. injection of NB-19 cells. Because osteoclastic bone resorption has been reported to play important roles in osteolysis in some cancers such as breast cancer, we next examined the osteoclast (OC)-inducing activity of NB-19 cells using a coculture system in which NB-19 cells were cultured with murine bone marrow cells containing OC precursors and stromal cells. NB-19 cells induced tartrate-resistant acid phosphatase-positive multinucleated OC-like cells without requirement of 1,25-dihydroxyvitamin D3 or other osteoclastogenic stimulators. To investigate the factors involved in the osteoclastogenesis in the coculture of mouse marrow cells and NB-19 cells, we performed reverse transcription-PCR analysis and revealed the increased expression of receptor activator of nuclear factor kappaB ligand (RANKL) in the coculture compared with the culture of bone marrow cells alone. Interleukin-1alpha and cyclooxygenase-2 expression in the murine marrow cells was also increased in the presence of NB-19 cells. To further study the role of RANKL in the OC-like cell formation in the coculture of NB-19 cells and murine marrow cells, an expression vector encoding the active portion of the murine osteoprotegerin, which is the native inhibitor of RANKL action, was constructed and introduced into COS-7 cells. The conditioned media of the COS-7 cells transfected with the osteoprotegerin expression vector effectively blocked OC-like cell formation in the coculture of the bone marrow cells and NB-19 cells. These results suggested that in the bone microenvironment of NB-19-bearing mi

Topics: Acid Phosphatase; Animals; Bone Marrow Cells; Bone Neoplasms; Carrier Proteins; Cell Communication; Coculture Techniques; COS Cells; Cyclooxygenase 2; Female; Glycoproteins; Humans; Interleukin-1; Isoenzymes; Membrane Glycoproteins; Membrane Proteins; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Neuroblastoma; Osteoclasts; Osteoprotegerin; Prostaglandin-Endoperoxide Synthases; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Receptors, Cytoplasmic and Nuclear; Receptors, Tumor Necrosis Factor; Recombinant Proteins; Reverse Transcriptase Polymerase Chain Reaction; Tartrate-Resistant Acid Phosphatase; Transplantation, Heterologous; Tumor Cells, Cultured

The tauopathies, which include Alzheimer's disease (AD) and frontotemporal dementias, are a group of neurodegenerative disorders characterized by filamentous Tau aggregates. That Tau dysfunction can cause neurodegeneration is indicated by pathogenic tau mutations in frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). To investigate how Tau alterations provoke neurodegeneration we generated transgenic mice expressing human Tau with four tubulin-binding repeats (increased by FTDP-17 splice donor mutations) and three FTDP-17 missense mutations: G272V, P301L, and R406W. Ultrastructural analysis of mutant Tau-positive neurons revealed a pretangle appearance, with filaments of Tau and increased numbers of lysosomes displaying aberrant morphology similar to those found in AD. Lysosomal alterations were confirmed by activity analysis of the marker acid phosphatase, which was increased in both transgenic mice and transfected neuroblastoma cells. Our results show that Tau modifications can provoke lysosomal aberrations and suggest that this may be a cause of neurodegeneration in tauopathies.

Topics: Acid Phosphatase; Animals; Cerebral Cortex; Cytoskeleton; Female; Hippocampus; Humans; Immunohistochemistry; Lysosomes; Male; Mice; Mice, Transgenic; Microscopy, Electron; Mutation; Neuroblastoma; Neurodegenerative Diseases; Neurons; Phosphorylation; Prosencephalon; tau Proteins; Tumor Cells, Cultured

Fetal Alz-50 clone 1 (FAC1) is a novel DNA binding protein with altered expression and subcellular localization during neuronal development and degeneration. FAC1 localizes to the cell body and neurites in undifferentiated neurons during development and in degenerating neurons during Alzheimer's disease progression. In the normal adult brain FAC1 is present predominantly in the nucleus of cortical neurons. When in the nucleus FAC1 has been shown to repress transcription by binding a specific DNA sequence. In the present study we demonstrate that the affinity of FAC1 for the identified DNA sequence is dramatically enhanced when FAC1 is phosphorylated. Phosphatase treatment of neuroblastoma nuclear extracts reduces FAC1 DNA binding affinity. Finally, inhibition of cellular serine/threonine phosphatases results in increased FAC1 DNA binding activity. These data suggest that FAC1 DNA binding activity is dependent upon and regulated by phosphorylation signals in the cell.

Topics: Acid Phosphatase; Adenosine Triphosphate; Animals; Antigens, Nuclear; Cell Nucleus; DNA; DNA-Binding Proteins; Humans; Nerve Tissue Proteins; Neuroblastoma; Neurons; Nuclear Proteins; Okadaic Acid; PC12 Cells; Phosphoprotein Phosphatases; Phosphorylation; Protein Binding; Rats; Recombinant Fusion Proteins; Response Elements; Transcription Factors; Tumor Cells, Cultured

Opioid receptor activity in neuroblastoma x glioma NG108-15 hybrid cell membranes was attenuated by acid phosphatase purified by high performance liquid chromatography and devoid of protease activity. Treatment of membranes with this phosphatase decreased opioid inhibition of adenylate cyclase and this effect was potentiated by the presence of the opioid agonist during the phosphatase treatment. Phosphatase treatment did not affect the number of opioid receptors but it did alter the distribution of receptors among affinity states, by increasing the percentage of receptors in the low affinity state. The similarities between these effects and desensitization of the opioid receptor, during chronic opioid treatment, are discussed.

Topics: Acid Phosphatase; Animals; Chromatography, High Pressure Liquid; Cyclic AMP; Diprenorphine; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Etorphine; Glioma; Guanfacine; Guanidines; Hybrid Cells; Neuroblastoma; Phenylacetates; Receptors, Opioid

The effect of prolonged exposure to ammonia on fluid-phase, receptor-mediated, and adsorptive (non specific) endocytosis in cultured neuroblastoma (Neuro-2a) cells were studied using fluorescein-labeled dextran, concanavalin A conjugated with fluorescein isothiocyanate, and cationized ferritin as tracers. Ammonia treatment increased the rate of endocytosis of cationized ferritin as well as the number of cell elements involved in the process. Moreover, the number of cytoplasmic components containing acid phosphatase activity was also found to increase following ammonia treatment. In contrast, flow-cytometric analyses showed that, under experimental conditions, exposure to ammonia did not alter the intralysosomal pH and had little effect on the fluid-phase and receptor-mediated endocytosis of fluorescein-labeled dextran and concanavalin-A fluorocrome, respectively.

Topics: Acetylcholinesterase; Acid Phosphatase; Ammonia; Animals; Concanavalin A; Dextrans; Endocytosis; Ferritins; Flow Cytometry; Fluorescein-5-isothiocyanate; Fluoresceins; Histocytochemistry; Hydrolyzable Tannins; Neuroblastoma; Time Factors; Tumor Cells, Cultured

Nine tumour lines with different developmental capacities were derived from spontaneous as well as from one induced teratocarcinoma: three teratocarcinoma-derived rhabdomyosarcomas TDR 602, TDR 694, and TDR 114; two teratocarcinoma-derived neuroblastomas TDN 2151 and TDN 2283; two teratocarcinoma-derived endodermal tumours TDE 274 and TDE 113; one multipotential teratocarcinoma OTT 2289, and one undifferentiated teratocarcinoma OTT 2158. Quantitative analyses of ten catabolic enzymes, i.e. alkaline and acid phosphatase, alpha- and beta-galactosidase, alpha- and beta-glucosidase, alpha-mannosidase, alpha-fucosidase, beta-glucuronidase, and hexosaminidase were carried out at the 20-cell level, and specific enzyme activity profiles were established for each of the tumour lines studied. These profiles may be used for the biochemical identification of a tumour type at the single cell level in addition to morphological and biological criteria.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Glycoside Hydrolases; Histocytochemistry; Kinetics; Mice; Mice, Inbred Strains; Neoplasms, Experimental; Neuroblastoma; Rhabdomyosarcoma; Teratoma

The nature of red fluorescent particles in vitally acridine orange stained C 1300 neuroblastoma monolayer cells was evaluated by electron microscopy, cytofluorometry, cytopharmacological and cell fractionation studies. At the ultrastructural level the distribution of red fluorescent granules correlated with that of the Golgi complex and Golgi derived structures during various stages of differentiation, mitosis, and under colcemid treatment. Cytopharmacological studies revealed that red fluorescence was displaced in a concentration and time dependent manner with the basic drugs chloroquine and quinacrine. Subcellular fractionation studies showed that acridine orange was concentrated in fractions that also contained the highest amount of acid phosphatase and electron dense vesicles. Vital acridine orange staining of neuroblastoma cells in culture can give information on the relationship between Golgi-derived vesicles and cell functions like proliferation and differentiation. The influence of drugs on these processes can be studied.

Topics: Acid Phosphatase; Acridine Orange; Animals; Cells, Cultured; Fluorescence; Fluorometry; Mice; Neoplasms, Experimental; Neuroblastoma; Staining and Labeling; Subcellular Fractions

Topics: Acid Phosphatase; Acridine Orange; Animals; Cell Differentiation; Histocytochemistry; Microscopy, Electron; Microscopy, Fluorescence; Neuroblastoma; RNA; Subcellular Fractions

Exposure of mouse neuroblastoma cells in culture to low pH for 8 days caused a sixfold increase in cell lipofuscin pigment. Pigment content was measured as the percent of cells having clumps of acid phosphatase-staining material. Hydergine, between 0.1 and 3 microgram/ml, caused a concentration-related decrease in pigment content. Hydergine also stimulated neurite formation in cells in regular pH medium and was slightly toxic to cells in low pH medium. These results support the use of neuroblastoma cells as an in vitro model for age pigment studies.

Topics: Acid Phosphatase; Aging; Animals; Cells, Cultured; Culture Media; Dihydroergotoxine; Lipofuscin; Mice; Models, Biological; Neoplasms, Experimental; Neuroblastoma; Pigments, Biological

Topics: Acid Phosphatase; Alkaline Phosphatase; Centrifugation, Density Gradient; Dopa Decarboxylase; Dopamine beta-Hydroxylase; Glucuronidase; Humans; Lysosomes; Microsomes; Mitochondria; Monoamine Oxidase; Neoplasm Proteins; Neuroblastoma; Norepinephrine; Subcellular Fractions; Tyrosine 3-Monooxygenase

Topics: Acid Phosphatase; Adenosine Triphosphatases; Alkaline Phosphatase; Bone Neoplasms; Carcinoma; Esterases; Glucuronidase; Histocytochemistry; Hodgkin Disease; Humans; Leukemia, Lymphoid; Leukemia, Myeloid; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Non-Hodgkin; Monoamine Oxidase; Multiple Myeloma; Neoplasm Metastasis; Neuroblastoma; Plasmacytoma; Sarcoma, Ewing