• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.1-8
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• 1996

Human cytomegalovirus (HCMV) replication and $Ca^{2+}$ response in human cell lines of neuronal origin were investigated. SK-N-SH (neuroblastoma cells) and A172 cells (glioblastoma cells) were used. SK-N-SH cells were permissive for HCMV multiplication with a delay of one day compared to virus multiplication in human embryo lung (HEL) cells. The delay of HCMV multiplication in SK-N-SH cells appeared to be correlated with a delay in the $Ca^{2+}$ response. The cytoplasmic free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) began to increase at 12 h p.i. in HCMV-infected SK-N-SH cells, while $[Ca^{2+}]_i$ increase in HCMV-infected HEL cells was observed as early as 3 h p.i. On the whole, the level of the increase in $[Ca^{2+}]_i$ in SK-N-SH cells was about 30% of that in HEL cells. On the other hand, in A172 cells infected with HCMV, neither production of infectious virus nor detectable increase in $[Ca^{2+}]_i$ was observed. Treatment with TPA of HCMV-infected SK-N-SH cells resulted in $[Ca^{2+}]_i$ increase at 6 h p.i. The stimulatory effect of TPA on HCMV- induced $[Ca^{2+}]_i$ increase continued until 12 h p.i., but TPA failed to stimulate the $Ca^{2+}$ response in SK-N-SH cells at 24 h p.i., suggesting that the effect of TPA had disappeared in SK-N-SH cells at that time point. In conclusion, SK-N-SH cells are permissive for HCMV replication and the delay in $Ca^{2+}$ response may be a consequence of the lower responsiveness of SK-N-SH cells than HEL cells to HCMV infection.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.196.2-197
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• 2003

Parkinson's disease (PO) is a widespread neurodegenerative disorder. Even though PD has been studied in many aspects, it is still unknown the molecular signaling mechanisms linking reactive oxygen species (ROS) and neuronal apoptosis in PD. A better understanding of cellular mechanisms that occur in Parkinson's disease is essential for development of new therapies. In this study we investigated the signaling molecules involved in neuronal apoptosis induced by 6-hydroxydopamine (6-OHDA) in human SK-N-SH neuroblastoma cells as a model cellular system. (omitted)

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.21
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• pp.9445-9451
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• 2014

Neuroblastoma is the most common extracranial solid tumor in children. Approximately half of the affected patients are diagnosed with high-risk poor prognosis disease, and novel therapies are needed. Sanguinarine is a benzophenanthridine alkaloid which has anti-microbial, anti-oxidant and anti-inflammatory properties. The aim of this study is whether sanguinarine has in vitro apoptotic effects and which apoptotic genes might be affected in the human neuroblastoma cell lines SH-SY5Y (N-myc negative), Kelly (N-myc positive, ALK positive), and SK-N-BE(2). Cell viability was analysed with WST-1 and apoptotic cell death rates were determined using TUNEL. After RNA isolation and cDNA conversion, expression of 84 custom array genes of apoptosis was determined. Sanguinarine caused cell death in a dose dependent manner in all neuroblastoma cell lines except SK-N-BE(2) with rates of 18% in SH-SY5Y and 21% in Kelly human neuroblastoma cells. Cisplatin caused similar apoptotic cell death rates of 16% in SH-SY5Y and 23% in Kelly cells and sanguinarine-cisplatin combinations caused the same rates (18% and 20%). Sanguinarine treatment did not affect apoptototic gene expression but decreased levels of anti-apoptotic genes NOL3 and BCL2L2 in SH-SY5Y cells. Caspase and TNF related gene expression was affected by the sanguinarine-cisplatin combination in SH-SY5Y cells. The expression of regulation of apoptotic genes were increased with sanguinarine treatment in Kelly cells. From these results, we conclude that sanguinarine is a candidate agent against neuroblastoma.

• Journal of Life Science
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• v.21 no.8
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• pp.1190-1198
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• 2011

Extraction methods of medicinal plants were evaluated for growth enhancing effects of human neuroblastoma SK-N-SH cells. Hot-water extraction (reflux for 5 hr), hot-water extraction post treatment (100$^{\circ}C$ or 120$^{\circ}C$, 90 min) and ethanol extraction (reflux for 5 hr) methods were applied to Angelica gigas, Rhemania glutinosa, Paeonia lactiflora and Cnidium officinale samples to extract their constituents. Cells were treated for 2 hr with various concentrations of extracts (0, 0.5, 1, 2, 4 ${\mu}g/{\mu}l}$ media) prior to $H_2O_2$ (250 ${\mu}M$) treatment for 2 hr to provide oxidative stress. Cell viability, caspase-3 expression and apoptosis were measured for cells treated with sample extracts. Hot-water extract exhibited a stronger growth enhancing and apoptosis protecting ability than other extracts. These activities were shown at less than 1 ${\mu}g/{\mu}l}$ concentration, and not greater than 2 ${\mu}g/{\mu}l}$ concentration. Hot-water extract contained more polyphenolic compounds than other extracts coming along with stronger antioxidant activity. The efficacy of antioxidant activity was stronger in the hot-water extract of Angelica gigas than other hot-water extracts of medicinal plants. These results suggest that hot-water extraction is an appropriate method to extract materials for growth enhancing and apoptosis protection of SK-N-SH cells, and hot-water extracts of Angelica gigas might be useful materials for protection from aging brain cells.

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.512-519
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• 2008

Three different polysaccharide-peptide complexes (PPC, named as Fr-I, Fr-II, and Fr-III) were produced by submerged mycelial culture of an entomopathogenic fungus Cordyceps sphecocephala, and their anticancer activities were investigated in human hepatocarcinoma (HepG2) and neuroblastoma (SK-N-SH) cells. The highest inhibitory effects of PPC on both HepG2 and SK-N-SH cells were achieved with Fr-I, whereas Fr-III with low molecular mass showed lower inhibition effects. Interestingly, the inhibitory effects of the three fractions were increased after protease digestion, suggesting that the inhibitory effects resulted mainly from the carbohydrate moiety, at least in the case of Fr-II and Fr-III, of PPC. The results of DNA fragmentation in PPC-induced apoptotic cells were confirmed by both DNA ladder assay and comet assay. Our investigation also showed that PPC-induced apoptosis of both cancer cells was associated with intracellular events including DNA fragmentation, activation of caspase-3, and modulation of Bcl-2 and Bax. We conclude that PPC has potential as a novel therapeutic agent for the treatment of both HepG2 and SK-N-SH cancer cells without any cytotoxicity against normal cells.

• The Korean Journal of Zoology
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• v.38 no.4
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• pp.542-549
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• 1995

The effect of extraceflular matrix (ECM) protein on the neuronai differentiation of SI(-N-SH and IMR-32 human neuroblastoma cell lines was examined. When ceils were cultured on the laminin/collagen coated plate for 7 days, the extensive neurite outgrowth was observed In IMR-32. To address the reason why IMR-32 cell llne did not respond to ECM proteins, the ECM mediated early signalling mechanisms were analysed in both SK-N-SH and IMR-32. When cells were plated on the laminin/collagen coated plates, tyrosine phosphorylated proteins were Increased within an hour In both of these cells. Moreover, the foaal adhesion IlInase (FAK) was tyrosine phosphorylated in both of these two cell lines. These results suggest that the ECM mediated early signalling mechanism was normal in IMR-32 cell line. The expression of both NSE and Bcl-2 was increased by ECM treatment in SK-N-SH. However, these components were not changed by ECM In IMR 32 cells to ECM component Is likely due to the abnomality of the transcriptional regulation mechanism which Is responsible for the neuronal differentiation.

• Journal of Ginseng Research
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• v.34 no.2
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• pp.138-144
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• 2010

Ginseng (Panax ginseng C.A. Meyer) has been shown to have anti-stress effects in animal studies. However, most studies have only managed to detect altered levels of biomarkers or enzymes in blood or tissue, and the actual molecular mechanisms by which ginseng exerts these effects remain unknown. In this study, the anti-oxidative effect of Korean red ginseng (KRG) was examined in human SK-N-SH neuroblastoma cells. Incubation of SK-N-SH cells with the oxidative stressor hydrogen peroxide resulted in significant induction of cell death. In contrast, pre-treatment of cells with KRG decreased cell death significantly. To elucidate underlying mechanisms by which KRG inhibited cell death, the expression of apoptosis-related proteins was examined by Western blot analysis. KRG pre-treatment decreased the expression of the pro-apoptotic gene caspase-3, whereas it increased expression of the anti-apoptotic gene Bcl-2. Consistent with this, immunoblot analysis showed that pre-treatment of the SK-N-SH cells with KRG inhibited expression of the pro-inflammatory gene cyclooxygenase 2 (COX-2). RT-PCR analysis revealed that the repression of COX-2 expression by KRG pre-treatment occurred at the mRNA level. Taken together, our data indicate that KRG can protect against oxidative stress-induced neuronal cell death by repressing genes that mediate apoptosis and inflammation.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.67-74
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• 2011

Objectives : The purpose of the study is to determine the neuroprotective effects of the water extract of Angelicae Acutilobae Radix(AA) on ischemia reperfusion-induced apoptosis in SK-N-SH human brain neuronal cells. Methods: SK-N-SH cells were treated with different concentrations of AA water extract (0.1, 0.2, 0.5 and 1.0 mg/ml) for 2 hr and then stimulated with Dulbecco's phosphate-buffered saline containing CI-DPBS: 3mM sodium azide and 10 mM 2-deoxy-D-glucose for 45 min, reperfused with growth medium, and incubated for 24 h. Cell viability was determined by WST-1 assay, and ATP/ADP levels were measured by ADP/ATP ratio assay kit. The levels of caspase-3 protein were determined by Western blot and apoptotic body was observed by Hoechst 33258 staining. Results : AA extract significantly inhibited decreasing the cell viability in ischemia-induced SK-N-SH cells. AA also increased the ratio of ADP/ATP in ischemia-induced neuronal cells and decreased the expression levels of apoptotic protein, caspase-3 and apoptotic DNA damage. Conclusions : Our results suggest that AA extract has a neuroprotective property via suppressing the apoptosis and increasing the energy levels in neuronal cells, suggesting that AA extract may has a therapeutic potential in the treatment of ischemic brain injury.

• The Korea Journal of Herbology
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• v.28 no.2
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• pp.45-53
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• 2013

Objectives : The purpose of the study is to determine the neuroprotective effects of the water and 80% EtOH extract of some herbal medicine plant on ischemia reperfusion-induced cell death in SK-N-SH human brain neuronal cells. Methods : SK-N-SH cells were treated with 3mM sodium azide and 10 mM 2-deoxy-D-glucose for 45 min, ptior to the addition of different concentrations of herbal medicine plant extract (0, 10, 25, 50, 100, 250, 500, 1000 ${\mu}g/ml$) for 2 hr and then reperfused with growth medium, incubated for 24 h. Cell viability was determined by WST-1 assay, and ATP/ADP levels were measured by ADP/ATP ratio assay kit. Results : Herbal medicine plant extract significantly inhibited decreasing the cell viability in ischemia-induced SK-N-SH cells. Also increased the ratio of ADP/ATP in ischemia-induced neuronal cells. Conclusions : Our results suggest that herbal medicine plant extract has a neuroprotective property via increasing the energy levels in neuronal cells, suggesting that extract may has a therapeutic potential in the treatment of ischemic brain injury. The exact component and mechanism remains for the future study.

• Preventive Nutrition and Food Science
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• v.10 no.4
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• pp.353-356
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• 2005

It has been recently reported that boron affects bone metabolism in humans and animals. In this study we examined whether boron affects the proliferation on various cell types, MG-63, HOS, Raw 264.7 and SK-N-SH. When treated with different concentrations of boron $(1,\;10,\;100{\mu}M)$ for 24 and 48 hr, the proliferation of MG-63 cells was enhanced at $10{\mu}M\;(p<0.05)$, for 24 hr. In HOS cells, boron had no effect on cell proliferation at 24 or 48 hr. In addition, treatment of pre-osteoclastic cells (Raw 264.7) with 1, 10, $100{\mu}M$ boron resulted in no effect on cell proliferation. Proliferation of neuronal cells (SK-N-SH) was enhanced by boron in a concentration dependent manner at low concentrations (0.1, 0.5, $1{\mu}M$). Besides proliferation activity, boron has an effect on the enhancement of NO production in SK-N-SH cells in a concentration-dependent manner. These studies showed that boron enhances proliferation of osteoblastic cells (especially MG-63), depending upon the concentration of boron. These results also provide further evidence of the positive effects of boron in neuronal disease.