• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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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)

• 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)

• Animal cells and systems
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• 제9권4호
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• pp.191-198
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• 2005

The effects of 6-aminonicotinamide (6-AN) on viability of IMR32 neuroblastoma cells in the presence of ATP or $NAD^+$ have been investigated. 6-AN caused marked reduction in cell viability and similar observations were also made with cells treated with 6-AN+ATP. However, cells treated with $6-AN+NAD^+$ showed cell viability similar to untreated cells. Morphologically, 6-AN and 6-AN+ATP treated cells showed loss of neurites, polyhedric shapes, shrinkage of cell bodies and formation of lysed cells, while $6-AN+NAD^+$ cells did not show any such changes. The flow cytometry analysis demonstrated that 6-AN increased cell population in $G_0/G_1$ phase and decreased cell population in Sand $G_2/M$ phase following a 72 h exposure. Western blot analysis showed that 6-AN stimulated a substantial increase in the level of the cdk inhibitor $p27^{kip1}$, but lowered the levels of cdk2, cyclin E and p-Rb. However, cdc25A and p53R2 were not significantly affected. Immunofluorscence staining of $p27^{kip1}$, cdk2, cyclin E and p-Rb revealed close correlation between the signal observed in the Western blot analysis. 6AN+ATP treated cells showed similar results obtained with 6-AN treated cells in expression of cdk2, cyclin E, p-Rb proteins and $p27^{kip1}$, $6-AN+NAD^+$ cells showed greater expression of cdk2, cyclin E and p-Rb than those in 6-AN and 6-AN+ATP treated cells. The results suggest that 6-AN induced the $G_0/G_1$ phase arrest in IMR32 neuroblastoma cell lines through the increase of $p27^{kip1}$ and the decrease of cdk2, cyclin E and p-Rb.

• 대한의생명과학회지
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• 제3권1호
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• pp.11-20
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• 1997

신경아세포종 세포 분화의 조절에 대한 연구는 아직 초기 단계에 불과하나 신경아세포종양의 임상적인 치료에 매우 중요한 기초가 된다. 본 연구는 신경아세포종 세포의 분화를 유도 할 수 있는 유용한 시약을 찾아내고자 하는 노력의 일환으로, 잘 알려진 DNA 합성 억제제가 신경 아세포종의 분화를 유도할 수 있는지를 살펴보고자 신경아세포종양 세포의 형태적, 생화학적 및 유전자 발현에 미치는 효과를 살펴보았다. 신경아세포종양으로부터 수립된 세포주, IMR-32 세포에 DNA 합성 억제제인 sodium butyrate, hydroxyurea, cytosine arabinoside를 각각 처리한 결과 처리하지 않은 대조군과는 달리 정상신경 세포 분화시 볼 수 있는 신경 돌기의 성장이 유도됨을 관찰할 수 있었으며 ,신경 전달 물질인 acetylcholine의 합성 효소인 choline acetyltransferase의 활성도가 현저히 증가되었다. 또한DNA합성 억제제를 처리하지 않은 IMR-32세포에서 탐지되지 않았던 c-myc 유전자의 발현이 시약 처리시 확연히 탐지됨을 관찰할 수 있었다. 이러한 실험 결과들은 DNA합성 억제제가 IMR-32세포의 성장을 억제하고 분화를 유도했음을 보여준 것이며, 어린 아이 시기에 많이 발병되는 신경아세포종양의 급속한 악성화나 전이의 억제기전을 제시해 줄 수 있으리라 생각한다.

• Journal of Korean Neurosurgical Society
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• 제42권5호
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• pp.392-399
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• 2007

Objective : Arsenic trioxide ($As_2O_3$) has been used as an anticancer agent in traditional Chinese medicine for thousand years and berberine is an isoquinoline alkaloid present that has indicated significant antimicrobial activity. We have examined the combined anticancer effects of $As_2O_3$ and berberine against the human neuroblastoma (HNB) SH-SY5Y cells in vitro, and to elucidate underlying molecular mechanism. Methods : HNB SH-SY5Y cells were treated with $2\;{\mu}M\;As_2O_3$ and $75\;{\mu}g/ml$ berberine, and their survival, cell death mechanism as well as synergistic cytotoxic effects were estimated by using MTT assay, DAPI staining, agarose gel electrophoresis, flow cytometric analysis, and western blot analysis. Results : The combined treatment of two drugs also markedly decreased cell viability. The cytotoxic effects of two drugs were revealed as apoptosis characterized by chromatin condensation, DNA fragmentation, and the loss of mitochondrial membrane potential. The apoptotic cytotoxicity was accompanied by activation of caspase-3 protease as well as decreased the expression of Bcl-2, Bid, and Bcl-x/L. In addition, the cells treated with combination of two drugs also showed significantly increased intracellular reactive oxygen species levels and lipid peroxidation compared to cells $As_2O_3$or berberine only. Conclusion : Combined treatment of $As_2O_3$ with berberine induced activation of apoptotic signaling pathways in HNB SH-SY5Y cells. These results suggest that the possibility of the combined treatment of two chemotherapeutic agents with low concentration improving cytotoxic effect for cancer cells with minimal side effects.

• 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.

• The Korean Journal of Physiology and Pharmacology
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• 제13권4호
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• pp.281-285
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• 2009

Rotenone, a mitochondrial complex I inhibitor, can induce the pathological features of Parkinson's disease (PD). In the present study, naringin, a grapefruit flavonoid, inhibited rotenone-induced cell death in human neuroblastoma SH-SY5Y cells. We assessed cell death and apoptosis by measuring mitogen-activated protein kinase (MAPKs) and caspase (CASPs) activities and by performing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Naringin also blocked rotenone-induced phosphorylation of Jun NH2-terminal protein kinase (JNK) and P38, and prevented changes in B-cell CLL/lymphoma 2 (BCL2) and BCL2-associated X protein (BAX) expression levels. In addition, naringin reduced the enzyme activity of caspase 3 and cleavages of caspase 9, poly (ADP-ribose) polymerase (PARP), and caspase 3. These results suggest that naringin has a neuroprotective effect on rotenone-induced cell death in human neuroblastoma SH-SY5Y cells.

• Clinical and Experimental Pediatrics
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• 제46권7호
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• pp.702-709
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• 2003

목 적 : $IFN{\gamma}$는 다양한 암세포에서 $TNF{\alpha}$와 FAS/CD95 수용체 발현을 증가시키거나 caspase나 Bcl-2 가족의 활성화를 조절하여 $TNF{\alpha}$와 FAS/FASL 유도성 세포고사를 촉진한다. 신경모세포종에서 $IFN{\gamma}$와 $TNF{\alpha}$는 협동적으로 세포 분화를 유도하거나 성장 억제를 일으킨다. 또한 일부 신경모세포종에서 자연적인 FAS 수용체 발현에도 불구하고 그 리간드 자극에 의한 세포고사 유도에는 실패하였고 $IFN{\gamma}$ 투여로 이를 극복할 수 있음이 보고되었다. 본 연구에서는 $IFN{\gamma}$가 $TNF{\alpha}$나 길항적 FAS/CD95 항체 유도성 세포고사를 촉진할 수 있는지 여부를 다양한 항암제에 대한 내성을 가지고 있는 신경모세포종 세포주를 이용하여 알아보았다. 방 법 : CHLA-15, CHLA-90와 LA-N-2 신경모세포종 세포주를 IMDM 배지로 배양하였고 유전자 재조합 $IFN{\gamma}$, $TNF{\alpha}$, 길항적 FAS/CD95 항체(CH-11)를 투여하였다. 세포 생존율은 형광기질인 calcein-AM을 이용한 DIMSCAN을 통하여 측정하였고, 세포고사 정도는 Annexin V-PE와 7-ADD염색을 이용한 유식세포 분석기를 통하여 분석하였고 pancaspase and caspase-8 억제 실험을 통하여 확인하였다. TNF와 FAS/CD95 수용체 표현은 각각에 대한 단클론 항체와 PE가 결합된 이차 항체를 이용하여 유식세포 분석기로 알아보았다. 결 과 : $IFN{\gamma}$ 또는 $TNF{\alpha}$ 단독 투여로는 모든 세포주에서 의의있는 세포 독성을 유도하지 못 했으나 $IFN{\gamma}$와 $TNF{\alpha}$을 병행 투여시에는 CHLA-15과 CHLA-90 세포주에서 의의있는 세포 생존율 감소와 공통 capase경로를 통한 세포고사를 협동적으로 촉진하였다. 또한 길항적 FAS/CD95 항체 단독 투여 시에는 모든 세포주에서 세포 생존율의 변화가 없었으나 $IFN{\gamma}$ 전 처치 후 투여 시에는 CHLA-90 세포주에서 현저한 세포 생존율 변화 및 세포고사를 유도하였다. $INF{\gamma}$ 치료 후 TNFRI와 FASR의 발현이 모든 세포주에서 현저히 증가하였는데 이는 일부 감수성이 있는 신경모세포종에서 $INF{\gamma}$에 의한 $TNF{\alpha}$와 FAS/CD95수용체 유도성 세포고사 촉진의 한 기전이 될 것으로 사료된다. 결 론: 일부 신경모세포종에서 $IFN{\gamma}$가 $TNF{\alpha}$와 길항적 FAS/CD95 항체 유도성 세포고사를 감작화 시켰으며 이는 수용체 발현의 증가와 동반되었다.

• 한국미생물·생명공학회지
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• 제25권1호
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• pp.102-105
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• 1997

In cultivating human neuroblastoma cells maximum number of neurites per cell and length of the neurite were estimated as 5.5 and 2.2 (nm), respectively It was found that there was correlation between growth and differentiation of nerve cells. Maximum specific BDNF production rate was also calculated as 2.5$\times $10$^{-5}$(ng/cell/day) at 7$\times $ 10$^{5}$ (viable cells/ml) of maximum cell density, corresponding to 100 (ng/mL) of BDNF. The secretion of BDNF was occurred most in the later peroids of the cultivation, yielding 75 (ng/mL) of BDNF. The production of rate of BDNF was elongated in adding 1 ($\mu $g/mL) of BDNF as well as 40% increase of the length of the BDNF. It proves that BDNF can be used as one of biopharmaceuticals to treat age-related diseases such as Alzheimer's disease and Prakinson's disease. It can also provide the information of scaling-up mammalian cell cuture system to economically produce BDNF.

• BMB Reports
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• 제36권4호
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• pp.354-358
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• 2003

For better understanding of functions of the Calcyclin Binding Protein (CacyBP) and exploring its possible roles in neuronal differentiation, the subcellular localization of human CacyBP was examined in retinoic acid(RA)-induced and uninduced neuroblastoma SH-SY5Y cells. Immunostaining indicated that CacyBP was present in the cytoplasm of uninduced SH-SY5Y cells, in which the resting $Ca^{2+}$ concentration was relatively lower than that of RA-induced cells. After the RA induction, immunostaining was seen in both the nucleus and cytoplasm. In the RA-induced differentiated SH-SY5Y cells, CacyBP was phosphorylated on serine residue(s), while it existed in a dephosphorylated form in normal (uninduced) cells. Thus, the phosphorylation of CacyBP occurs when it is translocated to the nuclear region. The translocation of CacyBP during the RA-induced differentiation of SH-SY5Y cells suggested that this protein might play a role in neuronal differentiation.