• 생명과학회지
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• 제21권8호
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• pp.1190-1198
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• 2011

한약재의 추출방법에 따른 인체신경모세포 SK-N-SH에 대한 보호 효과를 연구하였다. 당귀, 건지황, 작약 및 천궁을 시료로 사용하였고, 열수추출(환류냉각, 5 시간), 증숙추출(100$^{\circ}C$ 및 120$^{\circ}C$, 90분 후 열수추출) 방법과 에탄올추출(환류냉각, 5 시간)방법을 비교하였다. 추출물을 농도별로 SK-N-SH 세포에 2 시간 처리한 후 $H_2O_2$로 250${\mu}M$로 2 시간 산화스트레스를 유발한 다음 세포독성 및 apoptosis와 caspase-3의 발현 정도를 측정하였다. 모든 약재의 열수추출물이 다른 추출물보다 세포 증식을 촉진하였고, apoptosis를 억제하였다. 한약재 열수추출물 1${\mu}g/{\mu}l}$ 농도까지는 세포증식을 촉진하였지만, 그 이상의 농도에서는 오히려 감소시켰다. 열수추출물은 다른 추출물보다 총페놀성 화합물이 많이 함유되어 있었고, 항산화능이 높았다. 또한, 이와 같은 효과는 당귀의 열수추출물이 다른 약재 열수추출물보다 우수하였다. 본 연구결과는 약재의 열수추출법이 인체신경모세포인 SK-N-SH의 증식과 세포사멸 억제를 위해 가장 우수한 방법이었고, 당귀 열수추출물이 가장 우수한 효과를 지니고 있어 기억력 보호나 상실억제제로 활용할 수 있을 것이다.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2002년도 제9회 학술 발표회 프로그램과 논문초록
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• pp.46-46
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• 2002

Cdk5, a neuronal Cdc2-like kinase, exhibits a variety of functions in neuronal differentiation and neurocytoskeleton dynamics as well as neuronal degeneration and cell death. However, its role in retinoic acid (RA)-induced differentiation has not been reported yet. We newly found that RA treatment of SK-N-BE(2)C, human neuroblastoma, increased expression of Cdk5 concomitantly with a neuronal specific activator, p35.(omitted)

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.4007-4011
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• 2015

Objective: To explore the expression of RECK and relevant matrix metalloproteinases (MMPs) in hepatoblastoma (HB) and neuroblastoma (NB) and their clinical significance in the tumor metastasis. Materials and Methods: Forty-five wax-stone samples of HB and 43 wax-stone samples of NB removed by surgical resection and confirmed by pathology in Linyi Yishui Central Hospital were selected. According to presence and absence of metastasis, both NB and HB samples were divided into metastatic group and non-metastatic group, namely NB metastatic group (n=28), NB non-metastatic group (n=15), HB metastatic group (n=15) and HB non-metastatic group (n=30). The expression of RECK, membrane type-1 matrix metalloproteinase (MT1-MMP) in HB tissue and RECK, MMP-14 in NB tissue was detected using immunohistochemical method, and the correlation between RECK and MT1-MMP, MMP-14 was analyzed. Results: The metastatic rate of NB was dramatically higher than that of HB, with statistical significance (P=0.003). The positive rate of RECK expression in NB group (30.2%) was slightly lower than in HB group (40.0%), but no significant difference was presented (P=0.338). The positive rate of MMPs expression in NB metastatic group was evidently higher than in HB metastatic group (P=0.024). The results of Spearman correlation analysis revealed that the expression of RECK in HB and NB tissues had a significantly-negative correlation with MT1-MMP and MMP-14, respectively (r=-0.499, P=0.012; r=-0.636, P=0.000). Conclusions: In HB and NB tissues, RECK is expressed lowly, while relevant MMPs highly, and RECK inhibits the tumor invasion and metastasis through negative regulation of relevant MMPs.

• 대한생식의학회:학술대회논문집
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• 대한생식의학회 2007년도 제53차 추계학술대회 초록집
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• pp.115.2-116
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• 2007

• 대한미생물학회지
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• 제22권2호
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• pp.167-174
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• 1987

N-myc, a DNA sequence related to the oncogene c-myc, was found to be amplified in untreated primary neuroblastomas and the amplification appeared to be associated with advanced disease at diagnosis and rapid tumor progression. Synthetic peptides have been useful immunogens for generating antisera and monoclonal antibodies to a number of native proteins. In order to identify myc-related protein in the tumor cells, an antiserum against a synthetic hexapeptide (-Glu-Asp-Ile-Trp-Lys-Lys-), whose sequence corresponds to a part of the exon 2 of oncogene N-myc, was prepared by immunizing a rabbit with BSA-conjugated peptide. After ammonium sulfate precipitation and affinity column chromatography, it appeared to be specific to the peptide. Strong nuclear staining in immunoperoxidase method using this serum was observed in both human promyeloid leukemic cell line, HL-60(containing high c-myc copy number), and human neuroblastoma cell line, LA-N-5 (containing high N-myc copy number), whereas LA351 (human lymphoid cell line) cells did not react with the serum. This reaction was completely abrogated by incubating the antiserum with soluble excess peptide. These data suggest that the protein encoded by N-myc could be localized in the nucleus as c-myc protein and this antiserum can be used to detect myc-related tumor cells in clinical samples and to determine if the N-myc expression correlates with genomic amplification in cell lines, untreated primary tumors, and untreated metastases.

• 대한임상독성학회지
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• 제21권2호
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• pp.81-91
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• 2023

Purpose: Edible insect extracts have been used as an alternative source for medicinal supplements due to their significant antioxidative and anti-inflammatory activity. Recent studies have reported that anti-microbial peptides from insects have neuroprotective effects on dopamine toxins. The purpose of this study was to investigate the protective functions of mealworm (Tenebrio molitor) extract (MWE) on N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced cellular toxicity in SH-SY5Y neuroblastoma cells. Methods: Cellular toxicity induced by the MPTP toxin and the impact of MWE on cell survival were analyzed using MTT assays. DAPI staining was performed to observe apoptotic phenomena caused by MPTP. Changes in caspase-3 activity and protein expression were observed using enzyme activity assays and western blot assays, respectively. Results: MWE exerted significant antioxidant activity, which was measured by both DPPH and ABTS radical assays, with a dose-dependent relationship. Furthermore, MWE resulted in cellular proliferation in SHSY5Y cells in a dose-dependent manner. Furthermore, MWE pretreatment significantly inhibited MPTP-induced cytotoxicity, with a dose-dependent relationship. The morphological characteristics of apoptosis and increased reactive oxygen species induced by MPTP were also significantly reduced by MWE pretreatment. Conclusion: MWE treatment significantly attenuated MPTP-induced changes in the levels of proteins associated with apoptosis, such as caspase-3 and PARP. These findings suggest that MWE exerts neuroprotective effects on human neuroblastoma SH-SY5Y cells subject to MPTP-induced dopaminergic neurodegeneration.

• 대한한의학회지
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• 제28권1호통권69호
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• pp.198-210
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• 2007

Objectives : Nitric oxide(NO) is a reactive free radical and a messenger molecule in many physiological functions. However, excessive release NO of induces neurotoxicity. We investigated whether a mixture of red ginseng and paeonia radix prossesses a protective effect against sodium nitroprusside(SNP)-induced apoptosis in the human neuroblastoma cell line SK-N-MC. Methods : We performed 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, 4,6-diamidino-2-phenylindole(DAPD) staining, terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction(RT-PCR), Western blot analysis, and caspase-3 enzyme activity assay in SK-N-HFC cells. Result : MTT assay showed that SNP treatment significantly reduced the viabilities of cells and that pre-treatment with the red ginseng and paeonia radix mixture alleviated SNP-induced cytotoxicity. The cells treated with SNP exhibited several apoptotic features, while those pre-treated fir 1 h with the mixture of red ginseng and paeonia radix 1 h prior to SNP expose showed reduced apoptotic features. In addition, the cells pre-treated with the red ginseng and paeonia radix mixture for 1 h prior to SNP expose increased bel-2 expressions, decreased Bax expressions, and decreased caspase-3 enzyme activity. Conclusions : These results show that the red ginseng and paeonia radix mixture exerts a protective effect against SNP-induced apoptosis in SK-N-MC cells.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2003년도 제39회 학술심포지움
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• pp.77-77
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• 2003

• 대한한의학회지
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• 제31권3호
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• pp.55-65
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• 2010

Background: Nitric oxide (NO) is a reactive free radical gas and a messenger molecule. NO has many physiological functions, but excessive NO production induces neurotoxicity. Objective: The present study investigated whether the aqueous extract of Polygala tenuifolia Willdenow possesses a protective effect on NO-induced apoptosis in human neuroblastoma cell line SK-N-MC. Method: For this study, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and caspase-3 enzyme assay were performed. Result: Sodium nitroprusside (SNP) exposure significantly decreased the viability of cells. The cells treated with SNP exhibited several apoptotic features such as increasing of Bax expression, caspase-3 enzyme activity and inhibiting of Bcl-2 expression. On the other hand, the viability of cells pre-treated with the aqueous extract of Polygala tenuifolia Willdenow was increased dose-dependently. The cells pre-treated for 1 h with the aqueous extract of Polygala tenuifolia Willdenow followed by treatment with SNP showed a decreased occurrence of apoptotic features like decreasing Bax expressions, caspase-3 enzyme activity and increasing Bcl-2 expressions. The aqueous extract of Polygala tenuifolia Willdenow reduced apoptotic cell death in neuroblastoma cell line SK-N-MC through the inhibition of Bax-dependent caspase-3 activation and the increasing of Bcl-2 expression. Conclusion: Based on the present results, it is possible that Polygala tenuifolia Willdenow has therapeutic value for the treatment of a variety of NO-induced brain diseases.

• Journal of Microbiology and Biotechnology
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• 제18권3호
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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.