• Molecules and Cells
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• 제37권12호
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• pp.881-887
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• 2014

Cell proliferation is tightly controlled by the cell-cycle regulatory proteins, primarily by cyclins and cyclin-dependent kinases (CDKs) in the $G_1$ phase. The ankyrin repeat-rich membrane spanning (ARMS) scaffold protein, also known as kinase D-interacting substrate of 220 kDa (Kidins 220), has been previously identified as a prominent downstream target of neurotrophin and ephrin receptors. Many studies have reported that ARMS/Kidins220 acts as a major signaling platform in organizing the signaling complex to regulate various cellular responses in the nervous and vascular systems. However, the role of ARMS/Kidins220 in cell proliferation and cell-cycle progression has never been investigated. Here we report that knockdown of ARMS/Kidins220 inhibits mouse neuroblastoma cell proliferation by inducing slowdown of cell cycle in the $G_1$ phase. This effect is mediated by the upregulation of a CDK inhibitor p21, which causes the decrease in cyclin D1 and CDK4 protein levels and subsequent reduction of pRb hyperphosphorylation. Our results suggest a new role of ARMS/Kidins220 as a signaling platform to regulate tumor cell proliferation in response to the extracellular stimuli.

• 동의생리병리학회지
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• 제22권2호
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• pp.328-332
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• 2008

The purpose of this study was to identify the protective effect of Codonopis pilosula extract on cell death induced by $H_2O_2$ in SK-N-MC neuroblastoma cells. We measured the antioxidant effect by DPPH radical scavenging analysis, BSA analyssis and examined the cell viability by crystal violet and cytochrome C, Bax, Bcl-2, p53, p21 by using Western blot analysis. Codonopis pilosula extract scavenged DPPH radical in a dose-dependent manner and shown direct free radical scavenging effect, suggested that Codonopis pilosula extract have antioxidant effect in vitro. Treatment of cells with hydrogen peroxide, a reactive oxygen species, was to induce cell death and pretreatment with Codonopis pilosula extract attenuated the occurrence of $H_2O_2-induced$ cell death. To elucidate the protective mechanisms of action of Codonopis pilosula extract, Western blot analyses for Bcl-2 and Bax expression and cytochrome c release were carried out. Pretreatment with Codonopis pilosula extract induced the expression of Bcl-2 and suppressed the release of cytochrome c and Bax into the cytosol, thereby arresting $H_2O_2-induced$ apoptotic cell death. Especially p21 and p53 were decreased prior to $H_2O_2$ treatment. These results suggest that Codonopis pilosula extract is associated with the cell cycle and anti-apoptotic cell death.

• 대한한의학회지
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• 제21권4호
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• pp.183-192
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• 2000

Objectives : The present study was carried out to investigate the effects of Danchun-hwan(DCH) on the peroxynitrite-induced neural cell death in human neuroblastoma cell line, SH-SY5Y. Methods : The cultured cells were pretreated with DCH and exposed to 3-morpholinosydnonimine(SIN-1) that simultaneously generates NO and superoxide, thus possibly forming peroxynitrite. The cell damage was assessed by using MTT assay and crystal violet staining. Results : Exposure of the cells to SIN-1 for 24hr induced 75% apoptotic cell death, as evaluated by the occurrence of morphological nuclear changes characteristic of apoptosis using 4', 6-diamidino-2-phenylinole(DAPI). However, pretreatment of SH-SY5Y with the water extracts of DCH, inhibited the apoptotic cell death in a dose-dependent manner. DCH also inhibited SIN-1-induced apoptotic caspase 3-like protease activity in a dose-dependent manner. DCH recovered the depleted glutathione levels by SIN-1. Conclusions : Taken together, it is suggested that DCH protected human neuroblastoma cell line, SH-SY5Y, from the free radical injury mediated by peroxynitrite by a mechanism of elevating antioxidant, GSH.

• 한국기계가공학회지
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• 제12권4호
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• pp.10-15
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• 2013

In order to fabricate high sensitive and stable biosensors, we require the material with superior biocompatibility and physical-chemical stability. Many kinds of biomaterials have been evaluated to apply for bioindustry. Recently, carbon based diamond thin films have been focal pointed as bio-applications and their possibility has been evaluated. Diamond thin film has many advantages for electrochemical and biological applications, such as wide potential window (3.0-3.5V), low background current and chemical-physical stability. In this work, we have cultured neuroblastoma cell (SH-SY5Y) on the crystalline diamond films. We use MTT assay to evaluate the characteristic of cell culture on the substrates. As a result, neuroblastoma cell was cultured on the crystalline diamond film as similar as cell culture dish.

• 동의생리병리학회지
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• 제18권2호
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• pp.544-552
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• 2004

In oriental medicine, Radix Polygalae(RP) has been to treat tremors et al. But the mechanism how to decrease tremors was not known. The purpose of this study was to investigate the effect of RP on neurodegenerative disease. We used RP to execute the study of this defense mechanism on dopamine-induced cell death in human SH-SY5Y dopaminergic neuroblastoma cells. MTT assay was used to know the cytotoxicity of dopamine and the defense mechanism. As a result of this experiment, dopamine had cytotoxicity in human SH-SY5Y cells, but when it treated with RP, the cell survival rate increased. This suppressed the cell apoptosis, activation of caspase-3 protease, production of ROS, and repair of membrane potential change. In conclusion, RP has the protective effect on dopamine-induced cell death in human SH-SY5Y dopaminergic neuroblastoma cells, so this could be an effective agent on the neurodegenerative disease like Parkinsonism.

• Journal of Korean Neurosurgical Society
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• 제63권6호
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• pp.707-716
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• 2020

Objective : The function of B7H3, a member of the B7 family of proteins, in neuroblastoma (NB) remains poorly characterized. Here we examine the expression pattern of B7H3 in clinical NB specimens and characterize the phenotype of B7H3 knock-down in NB cell line. Methods : Immunohistochemical (IHC) staining was carried out to assess the expression of B7H3 in clinical NB specimens. Survival association was analyzed using five Gene Expression Omnibus (GEO) datasets (GSE85047, GSE45480, GSE62564, GSE16476, GSE49710). Clonogenic survival and flow cytometry were performed after B7H3 knockdown to assess the cellular proliferation and cell survival in vitro. Impact of B7H3 silencing on NB growth was examined in vivo using the SH-SY5Y xenograft model. Results : On IHC staining, B7H3 was widely expressed in clinical NB specimens. Analysis of the transcriptional profiles of five GEO datasets clinically annotated NB specimens revealed that decreased B7H3 expression was associated with improved overall survival. B7H3 knockdown suppressed the proliferation of the SH-SY5Y NB model in vitro and in vivo. Cell cycle analysis revealed that B7H3 silencing induced G1/S arrest. This arrest was associated with the suppression of E2F1 expression and induction of Rb expression. Conclusion : Our results demonstrate that B7H3 expression correlate with clinical survival in NB patients. Preliminary studies suggest that B7H3 may mediate the G1/S transition.

• Archives of Pharmacal Research
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• 제16권2호
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• pp.155-157
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• 1993

DNA transfection conditions were investigated by calcium phosphate-DNA co-precipitation in SK-N-BE(2)C human neuroblastoma cells. The DNA plasmid of TH2400CAT was used in which rat tyrosine hydroxylase gene was inserted into chloramphenicol acetyltransferase reporter gent. The transfection efficiency was 25-30% and the method was simple and reproducible. So, the method will be a good tool for transient transfection analysis.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.123.2-123.2
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• 2003

The cellular mechanisms by which excess exposure to the excitatory neurotransmitter glutamate can produce neuronal injury are unknown. In this study, we found that glutamate induced cell death at IC (50) of 100 microM on the cultured human SH-SY5Y neuroblastoma cells. It has been hypothesized that glutamate excitotoxicity is related with the elevation of calcium (Ca) levels. To determine the dependence of glutamate neurotoxicity on Ca environment, extracellular (EDTA) and intracellular (BAPTA/AM) chelator were used. (omitted)

• 생약학회지
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• 제35권2호통권137호
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• pp.105-109
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• 2004

The effect of MeOH extract of Cibotium barometz (or Cibaro) on nogo-A expression was studied by neurite cone collapse and neurite outgrowth assay. The degrees of mRNA expression of BDNF, GDNF, and Caspase-3 in nogo-A were also examined with SK-N-SH cell lines using RT-PCR and confocal microscopy methods. We have shown that Cibaro treatment inhibits nogo-A activation in SK-N-SH cell lines. It has been shown that Cibaro increases the expression rates of neurofilament and enhances neurite outgrowth in neuroblastoma cells as increasing the amount of Cibaro. It has been also shown that Cibaro increases the expression rates of BDNF, GDNF mRNA in neuroblastoma cells as increasing the amount of Cibaro. These results suggest that Cibaro induces neutrite outgrowth by nogo-A inactivation and is, therefore, crucial for the treatments against anaplastic disc and spinal neuronal anesthesia.

• Toxicological Research
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• 제16권2호
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• pp.133-139
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• 2000

This study was designed to evaluate the mechanism by which reactive nitrogen intermediates (RNI) induced the cytotoxicity of human doparminergic neuroblastoma SH-SY5Y cells. 3-Morpholino-sydnonimine (SIN-l), a donor of peroxynitrite (ONOO) and sodium nitroprusside (SNP), a donor of nitric oxide (NO) induced cell detachment and apoptotic death, as characterized by chromatin condensation, the ladder pattern fragmentation of genomic DNA and morphological nuclear changes. SIN-l also induced the activation of caspase 3-like protease in a time-dependent manner. Exogenous antioxidants, such as reduced glutathione (GSH), N-acetylcysteine (NAC), and selenium protected the cells from apoptotic death and reduced the activation of caspase 3-like protease by SIN-1. Furthermore, SIN-l directly reduced the intracellular levels of glutathione. Taken together, these data suggested that RNI including NO and peroxynitrite decrease the concentration of intracellular antioxidant such as GSH, which lead to the apoptotic death of human neuroblastoma SH-SY5Y cells.