Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Resistance to (2-chloroethyl)-3-sarcosinamide-1-nitrosourea (SarCNU), an experimental antitumor compound, was investigated in the sensitive SK-MG-1 cells and the 20-fold more resistant SKI-1 human glioma cells [which are 3-fold more resistant to 1,3,bis(2-chloroethyl)-1-nitrosourea (BCNU)]. The transport of SarCNU was examined by utilizing tritiated sarcosinamide. Sarcosinamide uptake into SK-MG-1 cells is via the catecholamine carrier that accommodates epinephrine. Dixon plot analysis of SarCNU inhibition of sarcosinamide uptake reveals that SarCNU is also accommodated by this carrier. The uptake of 0.5 mM [3H]sarcosinamide was temperature dependent, with similar levels of intracellular sarcosinamide accumulating at steady state in both cell lines. The uptake of sarcosinamide in SKI-1 cells obeyed Michaelis-Menten kinetics over a 200-fold range of concentrations with a Km of 1.52 +/- 0.151 mM and Vmax of 0.659 +/- 0.066 nmol/10(6) cells/min. This represents a more than 5-fold decrease in the uptake affinity and a more than 4-fold increase in the transport capacity compared with SK-MG-1 cells (Km = 0.282 +/- 0.041 mM; Vmax = 0.154 +/- 0.024 nmol/10(6) cells/min). The initial rate of sarcosinamide uptake is similar in both cell lines. Dixon plot analysis confirmed that SarCNU is a competitive inhibitor of sarcosinamide transport in SKI-1 cells with a Ki of 17.5 mM, which is more than 5-fold greater than the Ki obtained in SK-MG-1 cells. The steady state accumulation of SarCNU is significantly reduced by 47% in SKI-1 cells compared with the SK-MG-1 cells (cell to medium ratios of 0.65 +/- 0.11 and 1.22 +/- 0.08, respectively) (p less than 0.005). The accumulation of BCNU was comparable in the two cell lines. Since the Vmax of sarcosinamide (SarCNU) uptake is increased in the SKI-1 cells, the decrease in intracellular SarCNU is not related to decreased drug influx via the catecholamine carrier in SKI-1 cells. The efflux of tritiated sarcosinamide was temperature dependent and similar in both cell lines, with 54 and 58% of sarcosinamide being freely exchangeable in SKI-1 and SK-MG-1 cells, respectively. SarCNU efflux may or may not be altered. Since the expression of mdr is higher in the sensitive cells, it is unlikely that increased efflux of SarCNU mediated by the P-glycoprotein is responsible for drug resistance.(ABSTRACT TRUNCATED AT 250 WORDS)
Mol Pharmacol 1990 Sep
PMID:Mechanisms of resistance to (2-chloroethyl)-3-sarcosinamide-1-nitrosourea (SarCNU) in sensitive and resistant human glioma cells. 240 23

The role of membrane lipid composition in determining the electrical properties of neuronal cells was investigated by altering the available fatty acids in the growth medium of cultured neuroblastoma X glioma hybrid cells, clone NG108-15. Growth of the cells for several days in the presence of polyunsaturated fatty acids (linoleic, linolenic, and arachidonic) caused a pronounced decrease in the Na+ action-potential rate of rise (dV/dt) and smaller decreases in the amplitude, measured by intracellular recording. Oleic acid had no effect on the action potentials generated by the cells. In contrast, a saturated fatty acid (palmitate) and a trans monounsaturated fatty acid (elaidate) caused increases in both the rate of rise and the amplitude. No changes in the resting membrane potentials or Ca2+ action potentials of fatty acid-treated cells were observed. The membrane capacitance and time constant were not altered by exposure to arachidonate, oleate, or elaidate, whereas arachidonate caused a small increase in membrane resistance. Examination of the membrane phospholipid fatty acid composition of cells grown with various fatty acids revealed no consistent alterations which could explain these results. To examine the mechanism for arachidonate-induced decreases in dV/dt, the binding of 3H-saxitoxin (known to interact with voltage-sensitive Na+) channels was measured. Membranes from cells grown with arachidonate contained fewer saxitoxin binding sites, suggesting fewer Na+ channels in these cells. We conclude that conditions which lead to major changes in the membrane fatty acid composition have no effect on the resting membrane potential, membrane capacitance, time constant, or Ca2+ action potentials in NG108-15 cells. Membrane resistance also does not appear to be very sensitive to membrane fatty acid composition. However, changes in the availability of fatty acids and/or changes in the subsequent membrane fatty acid composition lead to altered Na+ action potentials. The primary mechanism for this alteration appears to be through changes in the number of Na+ channels in the cells.
Cell Mol Neurobiol 1985 Dec
PMID:The effects of exposure to exogenous fatty acids and membrane fatty acid modification on the electrical properties of NG108-15 cells. 241 16

1. The neuroblastoma x glioma hybrid NG108-15 cell line has been widely studied as a neuronal model for its serotonergic, cholinergic, and peptidergic properties. 2. The catecholamine and serotonin content and that of their major metabolites have been determined by high-performance liquid chromatography with electrochemical detection (HPLC-EC) in NG108-15 cells under differentiated and undifferentiated conditions. 3. Cellular contents of L-DOPA, norepinephrine, (NE), L-epinephrine (EPI), and dopamine (DA) in differentiated cells, induced by 1 mM dibutyryl cyclic AMP (dBcAMP), are 149, 40, 129, and 124%, respectively, higher than those in undifferentiated cells. 4. 3,4-Dihydroxyphenethylacetic acid (DOPAC), the major metabolite of DA, is detectable only in differentiated cells. Similarly, DOPAC is present only in culture medium from differentiated cells, and not that of undifferentiated cells. 5. Serotonin (5-HT) is detectable only in undifferentiated cells; and the level of 5-hydroxyindoleacetic acid (5-HIAA), the major metabolite of 5-HT, is also 12.7% higher is undifferentiated cells. 6. Comparative analyses of differentiated and undifferentiated cells in monolayer cultures and undifferentiated cells cultured in the presence of 1 mM dBcAMP under suspension conditions suggest that change in the indolamine content is due to cellular changes upon morphological differentiation. 7. The clonal NG108-15 cell line is also catecholaminergic, in addition to cholinergic and serotonergic; and a shift of neurotransmitter pattern from serotonin to dopamine production occurs during morphological differentiation.
Cell Mol Neurobiol 1989 Sep
PMID:Modification of the indolamine content in neuroblastoma x glioma hybrid NG108-15 cells upon induced differentiation. 248 34

Studies have established that major increases in muscarinic acetylcholine receptor (mAchR) binding in the brain appear to coincide with synaptogenesis. The neuroblastoma x glioma hybrid NG108-15 cell line has been demonstrated to possess numerous functional characteristics of intact neurons, including synapse formation with myotubes. The present study examines and characterizes the mAchR on the hybrid NG108-15 cells during differentiation, induced by 1 mM dBcAMP. Specific binding of [3H]-QNB for differentiated cells increases gradually to a final level of 130% (P less than 0.05) over the control undifferentiated cells during the first 24 hr of incubation. Further, this increase of receptor sites appears to correlate proportionately to the degree of neurite extension of the differentiating cells. The dissociation rate constant at equilibrium (Kd) and maximum binding capacity (Bmax) have been determined to be 5.6 nM and 920 fmol/10(6) cells, respectively, for differentiated cells, and 4.4 nM and 400 fmol/10(6) cells, respectively, for undifferentiated cells. Computer analyses of the data obtained from saturation experiments reveal a single class of binding sites for [3H]-QNB on both differentiated and undifferentiated cells. The Hill plot analysis of the QNB-binding indicates a Hill coefficient (nH) of 1.0 and 0.91 for differentiated and undifferentiated cells, respectively, suggesting the unity of receptor sites with no co-operativity. Our results depict that increases of mAchRs on intact cells correlate with the degree of cellular differentiation.
Mol Cell Biochem 1989 Apr 11
PMID:Characterization of muscarinic acetylcholine receptors on intact neuroblastoma x glioma NG108-15 cell upon induced differentiation. 254 90

1. The possibility that a long-lasting neuronal activation regulates the expression of muscarinic cholinergic receptors was studied with three cultured neuronal cell lines. 2. Continuous depolarization of a subclone of the neuroblastoma-glioma NG108-15 hybrid cells with potassium chloride increased by 45-75% the number of cholinergic muscarinic receptors, monitored with 3H-QNB, whereas a short incubation with KCl for 10 min or 6 hr had no effect. 3. The calcium channel blocker verapamil increased the effect of KCl. 4. Two cell lines, named SC9 and WC5, that originate from the rat brain, also bind 3H-QNB. They were therefore used to test whether the effect of chronic depolarization is universal. Depolarized SC9 and WC5 cells, in the presence or absence of verapamil, did not show an increased 3H-QNB binding. 5. Muscarinic receptors of both SC9 and WC5 cells have a higher affinity to pirenzepine than the M-3 receptor subtype of the neuroblastoma-glioma cells, suggesting therefore that the two rat brain cell lines possess M-1 or M-2 receptors. 6. The physiological significance of this differential role of depolarization on the expression of different muscarinic receptors is discussed in the context of their postreceptor second messengers.
Cell Mol Neurobiol 1989 Mar
PMID:Neuronal membrane depolarization and the control of cholinergic muscarinic receptors: selective effect on different neuronal cell types. 271 80

The presence of a brain tumor alters regional cerebral blood flow, oxygen consumption, and glucose utilization in adjacent and remote brain tissue, but its effect on brain neurotransmitter levels is unclear. In the present report, the levels of noradrenaline (NA), dopamine (DA), 5-hydroxytryptamine (5-HT), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 5-hydroxyindoleacetic acid (5-HIAA) in tumor tissue and gray and white matter obtained from cats with induced brain tumors were measured. Glioma cells (9L) were xenotransplanted into the central white matter of the right hemisphere, and 15 d later the brains were frozen in vivo. Samples of tumor, parietal (peritumor), temporal, and frontal gray and white matter were divided for analysis of water content and quantification of amines and their metabolites. The water content of white matter, but not gray matter, adjacent to the tumor was increased. Neurotransmitter amine and metabolite levels were much lower in the tumor than in brain tissue. In gray matter adjacent to the tumor, concentrations of DA and its metabolites HVA and DOPAC were significantly decreased from control, whereas 5-HIAA was increased. The NA, DA, HVA, and DOPAC levels were decreased in temporal gray matter, whereas all amine and metabolite levels were unchanged in frontal gray matter. These results indicate that altered neurotransmitter metabolism is one of the effects of the presence of a brain tumor.
Mol Chem Neuropathol 1989 Apr
PMID:Regional monoamine and metabolite levels in a feline brain tumor model. 274 38

Differential hybridization of a cDNA library from rat C6 glioma cells with cDNA probes from naive C6 glioma cells and from cells exposed to 17 beta-estradiol identified cDNAs of an mRNA stimulated by 17 beta-estradiol. This mRNA designated ESP1 mRNA, reached maximal levels after 8 h of treatment with 17 beta-estradiol. The stimulation was not suppressed by cycloheximide. Dexamethasone treatment of C6 glioma cells did not induce ESP1 mRNA. It codes for a 164 amino acids long peptide. The sequence is similar in part to that of CRIP protein, a probably member of the ferredoxin superfamily. The conservation of primary structure suggests a role of ESP1 peptide in oxygen consumption. ESP1 mRNA expression is sexually dimorphic in body tissue, whereas it is expressed to comparative levels in the brain of adult males and females. This suggests that 17 beta-estradiol stimulates the expression of the ESP1 gene in the brain of both gender.
Mol Cell Endocrinol 1989 Apr
PMID:Characterization of an estradiol-stimulated mRNA in the brain of adult male rats. 274 28

The amount and distribution of glial fibrillary acidic protein (GFAP) were determined by flow cytometry (FCM) and an enzyme-linked immunosorbent assay (ELISA) in five human glioma cell lines stained by the indirect immunofluorescence method using anti-GFAP monoclonal antibody. Standard reference beads containing a known amount of fluorescein were used to calibrate the flow cytometer; however, the intensity of fluorescence from these beads was too weak to allow direct comparison with the fluorescence from the stained cells. Therefore, the flow cytometer was recalibrated using reference beads with a fluorescence intensity similar to that of the glioma cells. By comparing the fluorescence intensities of the two types of reference beads, it was possible to determine the fluorescein content of the stained cells directly from the relative fluorescence intensity (channel number). Glioma cell lines 343 MGA, SF 126, SF 188, U 251, and U 87 had fluorescein concentrations of 72.0 +/- 6.8, 8.1 +/- 0.3, 52.6 +/- 3.1, 86.4 +/- 4.0, and 56.2 +/- 2.9 x 10(5) (mean +/- standard error) Eq Sol Mol (equivalent solution of mole), respectively. The GFAP content of these cell lines, determined by ELISA, was 15.7 +/- 5.2, 0.5 +/- 0.1, 11.1 +/- 2.0, 20.8 +/- 4.6, and 9.5 +/- 2.7 pg GFAP/cell, respectively, and correlated closely with the results of FCM (R = 0.983, p less than 0.0028). A linear regression analysis yielded the following equation: pg GFAP/cell = -2.3376 + 0.2518 x FCM integrated mean channel number (fluorescein concentration: 10(5) Eq Sol Mol).(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Quantitation and distribution analysis of glial fibrillary acidic protein in human glioma cells in culture. 276 8

The regulation and expression of protein kinase C (PKC) and phosphomyristin C (PMC) (a principal substrate of PKC which is the major myristylated protein in lymphocyte and glioma lines that express it) in murine B and T lymphocytes were investigated. Both PMC and PKC are differentially regulated during T-cell development. The level of PMC expression is highest in CD4-8-, intermediate in CD4+8+, and lowest in J11d-, CD4, or CD8 single-positive thymocytes. PKC is equally expressed by all three thymic populations. In striking contrast to thymocytes, resting peripheral lymph node T cells and T-cell clones express little if any PMC and reduced levels of PKC. Neither PKC nor PMC is significantly induced upon the activation of lymph node T cells: treatment with anti-CD3 antibodies or anti-CD3 and interleukin-2 fails to induce PKC, whereas PMC is not induced by anti-CD3 alone and is only slightly induced by anti-CD3 and interleukin-2. In contrast to the situation with T cells, PMC and PKC are constitutively expressed at moderate levels in mature B cells. PMC is greatly increased in B-cell blasts generated by cross-linking the antigen receptor with anti-immunoglobulin. These results demonstrate that PMC and PKC are differentially regulated during the development and activation of B and T cells, suggesting that cellular events that rely upon PKC and PMC may differ during ontogeny and activation of different lymphocyte subsets.
Mol Cell Biol 1989 Sep
PMID:A major myristylated substrate of protein kinase C and protein kinase C itself are differentially regulated during murine B- and T-lymphocyte development and activation. 278 36

Human cell lines with neuronal and neuroendocrine features were examined for their expression of pp60c-src, the cellular homolog of the transforming gene product pp60v-src of Rous sarcoma virus. Four neuroblastoma (LA-N-5, SH-SY5Y, Paju, and SK-N-MC) and three small-cell lung carcinoma (U-2020, U-1690, and U-1285) cell lines were selected on the basis of their stage of neurocrine differentiation, as determined by the expression of neuron-specific enolase. In an immune complex protein kinase assay, all seven cell lines displayed c-src kinase activity which was considerably higher than that found in nonneurocrine cells (human diploid fibroblasts, glioma, and non-small cell lung carcinoma cell lines). Furthermore, the c-src kinase activity, as determined by autophosphorylation or phosphorylation of an exogenous substrate, enolase, correlated with the stage of neurocrine differentiation. There was an approximately 30-fold difference in c-src kinase autophosphorylation activity between the cell lines representing the highest and lowest stages of neurocrine differentiation. A similar variation was found in the steady-state levels of the c-src protein of these cell lines. Highly differentiated neuroblastoma cells expressed two forms of the src protein. Digestion by Staphylococcus aureus V8 protease did reveal structural diversity in the amino-terminal ends of these c-src molecules. In summary, we found a clear correlation between c-src kinase activity and the stage of neuronal and neuroendocrine differentiation. Thus, the phenotypic similarity between neurons and neuroendocrine cells includes high c-src expression.
Mol Cell Biol 1987 Dec
PMID:Expression of c-src in cultured human neuroblastoma and small-cell lung carcinoma cell lines correlates with neurocrine differentiation. 283 Apr 84


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