Expression of trkA cDNA in neuroblastomas mediates differentiation in vitro and in vivo

1993 ◽  
Vol 13 (12) ◽  
pp. 7447-7456
Author(s):  
H Matsushima ◽  
E Bogenmann
Keyword(s):  
Cell Differentiation ◽  
Cell Line ◽  
Neuronal Cell ◽  
Human Neuroblastoma ◽  
Myc Oncogene ◽  
Ngf Receptor ◽  
Neuronal Cell Differentiation ◽  
Mature Neurons

The human trkA cDNA was transfected into a malignant human neuroblastoma (NB) cell line (HTLA230) to investigate its role in NB growth and differentiation. This cell line lacks expression of both endogenous trkA and gp75NGFR genes. Transfectants expressing the trkA mRNA and surface-bound receptors transcriptionally activate immediate-early genes (c-fos, c-jun, and jun-B) following nerve growth factor (NGF) stimulation. NGF treatment induces growth arrest as well as down-regulation of the amplified N-myc oncogene. Genes selectively expressed in mature neurons (SCG-10, ret proto-oncogene, GAP-43, etc.) are transcriptionally activated, and neurite outgrowth further demonstrates differentiation of transfectants following NGF stimulation. trkA-expressing NB cells remain tumorigenic in nude mice; however, subcutaneous treatment of tumor-bearing mice with NGF induces Schwannian and neuronal cell differentiation similar to the induction seen in human ganglioneuroblastomas. Thus, trkA expression in HTLA230 cells is sufficient to generate a functional NGF receptor complex that leads to growth-arrested and differentiated NB cells in vitro and in vivo in the presence of NGF. Hence, NGF may play a crucial role in NB cell differentiation and regression in vivo.

scholarly journals Expression of trkA cDNA in neuroblastomas mediates differentiation in vitro and in vivo.

1993 ◽  
Vol 13 (12) ◽  
pp. 7447-7456 ◽  
Author(s):  
H Matsushima ◽  
E Bogenmann
Keyword(s):  
Cell Differentiation ◽  
Cell Line ◽  
Neuronal Cell ◽  
Human Neuroblastoma ◽  
Myc Oncogene ◽  
Ngf Receptor ◽  
Neuronal Cell Differentiation ◽  
Mature Neurons

The human trkA cDNA was transfected into a malignant human neuroblastoma (NB) cell line (HTLA230) to investigate its role in NB growth and differentiation. This cell line lacks expression of both endogenous trkA and gp75NGFR genes. Transfectants expressing the trkA mRNA and surface-bound receptors transcriptionally activate immediate-early genes (c-fos, c-jun, and jun-B) following nerve growth factor (NGF) stimulation. NGF treatment induces growth arrest as well as down-regulation of the amplified N-myc oncogene. Genes selectively expressed in mature neurons (SCG-10, ret proto-oncogene, GAP-43, etc.) are transcriptionally activated, and neurite outgrowth further demonstrates differentiation of transfectants following NGF stimulation. trkA-expressing NB cells remain tumorigenic in nude mice; however, subcutaneous treatment of tumor-bearing mice with NGF induces Schwannian and neuronal cell differentiation similar to the induction seen in human ganglioneuroblastomas. Thus, trkA expression in HTLA230 cells is sufficient to generate a functional NGF receptor complex that leads to growth-arrested and differentiated NB cells in vitro and in vivo in the presence of NGF. Hence, NGF may play a crucial role in NB cell differentiation and regression in vivo.

MPTP-Induced Modulation of Neurotransmitters in SH-SY5Y Human Neuroblastoma Cells

1998 ◽  
Vol 17 (6) ◽  
pp. 677-701 ◽  
Author(s):  
Xiaoou Song ◽  
Marion Ehrich
Keyword(s):  
Cell Line ◽  
Neuroblastoma Cells ◽  
Neuronal Cell ◽  
Cholinergic Antagonists ◽  
Mao Inhibitor ◽  
Human Neuroblastoma ◽  
Hydroxyindoleacetic Acid ◽  
Mao Activity

Neurotoxic effects of MPTP(1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine) were evaluated in vitro using a human neuronal cell line, SH-SY5Y, that contained features contributing to expression of MPTP toxicity in vivo, namely, a transport system for dopam ine (DA) and monam ine oxidase (MAO) activity. In this model system, MPTP was found to reduce levels of catecholamines (DA, norepinephrine, epinephrine), serotonin (5-HT), and the 5-HT metabolite 5-hydroxyindoleacetic acid (5-HIAA). MPTP enhanced 3H-DA release, which could contribute to the reduction in DA concentrations seen in these cells. In addition, MPTP inhibited MAO activity (Ki 2.26 X 10-5 M). Pretreatment with the MAO inhibitor pargy-line protected the cells from MPTP-induced alterations of catecholamines and the decrease in 5-HT. In this in vitro model, the cholinergic antagonists atro-pine and A-tubocurarine also protected cells from MPTP-induced alterations of catecholamines. The capability of cholinergic antagonists to prevent the MPTP-induced alterations of catecholamine concentrations suggests a possible cholinergic contribution to MPTP neurotoxicity in this cell line.

Abstract 2357: Src Kinase Inhibition Blocks Thrombin-induced Brain Injuries without Cognitive Side Effects

Stroke ◽  
2012 ◽  
Vol 43 (suppl_1) ◽  
Author(s):  
Da-Zhi Liu ◽  
Bradley P Ander ◽  
Ali Izadi ◽  
Ken Van ◽  
Xinhua Zhan ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cognitive Deficits ◽  
Neuronal Cell ◽  
Thrombin Injection ◽  
Cell Cycle Reentry ◽  
Cell Cycle Inhibition ◽  
Mature Neurons ◽  
Cognitive Side Effects

Intracerebral hemorrhage (ICH) activates thrombin, a potent mitogen. Thrombin triggers mitosis by modulating several intracellular mitogenic molecules including Src family kinases. These molecules regulate mitogen-activated protein kinases (MAPKs) and cell cycle proteins such as cyclin-dependent kinases (Cdks); and play critical roles in mitogenic signaling pathways and cell cycle progression. Since aberrant cell cycle reentry results in death of mature neurons, cell cycle inhibition appears to be a candidate strategy for the treatment of neurological diseases including ICH. However, this can also block cell cycle (proliferation) of neural progenitor cells (NPCs) and thus impair brain neurogenesis leading to cognitive deficits. We hypothesized that inhibition of cell cycle by blocking mitogenic signaling molecules (i.e., Src family kinase members) blocks cell cycle reentry of mature neurons without injuring NPCs, which will avoid cognitive side effects during cell cycle inhibition treatment for ICH. Our data shows: (1) Thrombin 30U/ml results in apoptosis of mature neurons via neuronal cell cycle reentry in vitro ; (2) PP2 (Src family kinase inhibitor) 0.3 µM attenuates the thrombin-induced neuronal apoptosis via blocking neuronal cell cycle reentry, but does not affect the viability of NPCs at the same doses in vitro ; (3) Intracerebral ventricular thrombin injection (20U, i.c.v.) results in neuron loss in hippocampus and cognitive deficits 5 weeks after thrombin injection in vivo ; (4) PP2 (1mg/kg, i.p.), given immediately after thrombin injection (i.c.v.), blocks the thrombin-induced neuron loss in hippocampus and cognitive deficits, whereas PP2 on its own at the same doses does not affect normal cognition in vivo . These suggest that Src kinase inhibition prevents hippocampal neuron death via blocking neuronal cell cycle reentry after ICH, but does not affect survival of NPCs.

Vitamin D as Radiosensitizer: A Review in Cell Line -

2020 ◽  
Vol 10 (6) ◽  
pp. 315-324
Author(s):  
Fahmi Radityamurti ◽  
Fauzan Herdian ◽  
Tiara Bunga Mayang Permata ◽  
Handoko Handoko ◽  
Henry Kodrat ◽  
...  
Keyword(s):  
Vitamin D ◽  
Cell Differentiation ◽  
Cell Line ◽  
Cell Lines ◽  
Anticancer Agent ◽  
Medical Literature ◽  
In Vitro Studies ◽  
Anti Cancer

Introduction: Vitamin D has been shown to have anti-cancer properties such as antioxidants, anti-proliferative, and cell differentiation. The property of vitamin D as an anticancer agent triggers researchers to find out whether vitamin D is useful as a radiosensitizer. Multiple studies have been carried out on cell lines in various types of cancer, but the benefits of vitamin D as a radiosensitizer still controversial. This paperwork aims to investigate the utilization of Vitamin D3 (Calcitriol) as radiosensitizer in various cell line through literature review.Methods: A systematic search of available medical literature databases was performed on in-vitro studies with Vitamin D as a radiosensitizer in all types of cell lines. A total of 11 in-vitro studies were evaluated.Results: Nine studies in this review showed a significant effect of Vitamin D as a radiosensitizer agent by promoting cytotoxic autophagy, increasing apoptosis, inhibiting of cell survival and proliferation, promoting gene in ReIB inhibition, inducing senescene and necrosis. The two remaining studies showed no significant effect in the radiosensitizing mechanism of Vitamin D due to lack of evidence in-vitro settings.Conclusion: Vitamin D have anticancer property and can be used as a radiosensitizer by imploring various mechanism pathways in various cell lines. Further research especially in-vivo settings need to be evaluated.

scholarly journals Inhibition of the SK-N-MC human neuroblastoma cell line in vivo and in vitro by a novel nutrient mixture

2013 ◽  
Vol 29 (5) ◽  
pp. 1714-1720 ◽  
Author(s):  
M. WAHEED ROOMI ◽  
TATIANA KALINOVSKY ◽  
NUSRATH W. ROOMI ◽  
ALEKSANDRA NIEDZWIECKI ◽  
MATTHIAS RATH
Keyword(s):  
Cell Line ◽  
Neuroblastoma Cell ◽  
Neuroblastoma Cell Line ◽  
Human Neuroblastoma Cell ◽  
Human Neuroblastoma ◽  
Human Neuroblastoma Cell Line

scholarly journals The Mixture of Gotu Kola, Cnidium Fruit, and Goji Berry Enhances Memory Functions by Inducing Nerve-Growth-Factor-Mediated Actions Both In Vitro and In Vivo

Nutrients ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 1372
Author(s):  
Jin Gyu Choi ◽  
Zahra Khan ◽  
Seong Min Hong ◽  
Young Choong Kim ◽  
Myung Sook Oh ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Neuronal Cell ◽  
Main Role ◽  
Memory Functions ◽  
Nerve Growth ◽  
Goji Berry

Nerve growth factor (NGF), a typical neurotrophin, has been characterized by the regulation of neuronal cell differentiation and survival involved in learning and memory functions. NGF has a main role in neurite extension and synapse formation by activating the cyclic adenosine monophosphate-response-element-binding protein (CREB) in the hippocampus. The purpose of this study was to determine whether a mixture of Gotu Kola, Cnidium fruit, and Goji berry (KYJ) enhances memory function by inducing NGF-mediated actions both in vitro and in vivo. The KYJ combination increased NGF concentration and neurite length in C6 glioma and N2a neuronal cells, respectively. Additionally, we discovered memory-enhancing effects of KYJ through increased NGF-mediated synapse maturation, CREB phosphorylation, and cell differentiation in the mouse hippocampus. These findings suggest that this combination may be a potential nootropic cognitive enhancer via the induction of NGF and NGF-dependent activities.

Abstract W P248: miRNA Profiling of the Ischaemic Penumbra

Stroke ◽  
2015 ◽  
Vol 46 (suppl_1) ◽  
Author(s):  
Emily N Ord ◽  
Chris McCabe ◽  
Catriona McDonald ◽  
John D McClure ◽  
I M Macrae ◽  
...  
Keyword(s):  
Cell Line ◽  
Neuronal Cell ◽  
Mrna Translation ◽  
In Vitro Study ◽  
Brain Regions ◽  
Artery Occlusion ◽  
Paramagnetic Resonance ◽  
Spontaneously Hypertensive

MicroRNAs (miRNAs) are small non-coding RNA molecules (20 - 24 nucleotides) that inhibit mRNA translation. Demonstrated to have key roles in normal CNS development & function, they have also emerged to have effecter roles in the pathogenesis & endogenous repair mechanisms following stroke. To select 2 miRNAs to modulate therapeutically we profiled miRNA expression of the evolving (24h) & final (72h) peri-infarct tissue of adult spontaneously hypertensive stroke-prone rats (SHRSP) following 45 min transient middle cerebral artery occlusion (tMCAO). T2-weighted magnetic resonance imaging (MRI) was used for accurate dissection of the peri-infarct tissue, with equivalent brain regions taken from time-matched shams (n=6/group). Of the 754 miRNAs evaluated (TaqMan® human miRNA microarray card v3.0 Applied Biosystems) 89 were determined as differently regulated following tMCAO. 22 of these miRNAs were relevant in stroke & were thus validated by Taqman® qRT-PCR using specific probes (n=9 /group). 5 miRNAs were successfully validated; miR-34b & miR-520b were selected as miRNAs of interest due to their novelty, time of endogenous regulation & targets. An in vitro study to determine whether upregulation/knock-down of these miRNAs would demonstrate functional effects in classical hypoxic pathways was performed. A rat neuronal cell line (B50) & glial cell line (B92) were subjected to 9hr hypoxia (1% O2 -serum) & 24h reoxygenation (+serum) +/- miR-34b or miR-520b regulation. Upregulation of either miRNA in B50 cells demonstrated a reduction in apoptosis, assessed qualitatively by Caspase-3 immunocytochemistry & quantitatively by cell death detection ELISA (p<0.01 vs hypoxic non-treated cells (NTC)). Upregulation of either miRNA in B92 cells significantly reduced superoxide production, assessed by electron paramagnetic resonance (p<0.001 vs hypoxic NTC). MiR-520b significantly lowered levels of lipid peroxidation in B92 cells, assessed by malondialdehyde assay, & both were significantly effective in B50 cells (p<0.01 vs hypoxic NTC). These data suggest miR-34b & -520b upregulation ameliorates damage following hypoxia/reperfusion in cerebral cell lines. Future studies will assess the effect of modulating these miRNAs in vivo.

In Vitro Labeling Strategies for Identifying Primary Neural Tissue and a Neuronal Cell Line after Transplantation in the Cns

1993 ◽  
Vol 2 (2) ◽  
pp. 131-149 ◽  
Author(s):  
Stephen M. Onifer ◽  
Linda A. White ◽  
Scott R. Whittemore ◽  
Vicky R. Holets
Keyword(s):  
Escherichia Coli ◽  
Cell Line ◽  
Neuronal Cell ◽  
Fluorescein Isothiocyanate ◽  
Initial Step ◽  
Host Cells ◽  
Lacz Gene ◽  
Raphe Neurons

Potential labels for identifying embryonic raphe neurons and a clonal, neuronally differentiating, raphe-derived cell line, RN33B, in CNS transplantation studies were tested by first characterizing the labels in vitro. The labels that were tested included 4,6-diamidino-2-phenylindole hydrochloride, 1,1′-dioctadecyl-3,3,3′-tetramethylindocarbocyanine perchlorate, the Escherichia coli lacZ gene, Fast Blue, Fluoro-Gold, fluorescein-conjugated latex microspheres, fluorescein isothiocyanate-conjugated or nonconjugated Phaseolus vulgaris leucoagglutinin, methyl o-(6-amino-3′-imino-3H-xanthen-9-yl) benzoate monohydrochloride, or tetanus toxin C fragment. Subsequently, the optimal in vitro labels for embryonic raphe neurons and for RN33B cells were characterized in vivo after CNS transplantation. In vitro, 1,1-dioctadecyl-3,3,3′-tetramethylindocarbocyanine perchlorate (DiI) optimally labeled embryonic neurons. The Escherichia coli lacZ gene optimally labeled RN33B cells. Most labels were rapidly diluted in cultures of embryonic astrocytes and proliferating RN33B cells. Some labels were toxic and were often retained in cellular debris. In vivo, DiI was visualized in transplanted, DiI-labeled raphe neurons, but not in astrocytes up to 1 mo posttransplant. DiI-labeled host cells were seen after transplantation of lysed, DiI-labeled cells. β-Galactosidase was visualized in transplanted, Escherichia coli lacZ gene-labeled RN33B cells after 15 days in vivo. No β-galactosidase was seen in host cells after transplantation of lysed, lacZ-labeled RN33B cells. The results demonstrate that labels for use in CNS transplantation studies should be optimized for the specific population of donor cells under study, with the initial step being characterization in vitro followed by in vivo analysis. Appropriate controls for false-positive labeling of host cells should always be assessed.

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