• The Journal of Korean Medicine
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• v.26 no.4
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• pp.12-21
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• 2005

Objectives: In the present study, we investigated whether an aqueous extract of Armeniacae semen induces apoptotic neuronal cell death upon mouse N2a neuroblastoma cells. Methods: 1. Cell viability was determined by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTI) assay. 2. For in situ detection of apoptotic cells, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, 4,6-diamidino-2-phenylindole (DAPI) staining. 3. The fraction of cells was revealed by flow cytometric analysis used that. 4. For detection of apoptotic DNA cleavage, DNA fragmentation assay was performed. 5. For detection of bax and bcl-2, Western blot analysis was performed. 6. Caspase enzyme activity was measured using caspase-3 assay. Results: From the present results, N2a neuroblastoma cells treated with Armeniacae semen extract exhibited several characteristics of apoptosis. A treatment of Armeniacae semen extract was shown to increase the expression of Bax, a proapoptotic protein, and the treatment decreased the expression of Blc2, an anti-apoptotic protein. In addition, Armeniacae semen extract increased the caspase-3 enzyme activity. Conclusions: The present results show that Armeniacae semen extract induces apoptotic cell death in mouse N2a neuroblastoma cells.

• 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.

• The Korea Journal of Herbology
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• v.21 no.4
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• pp.77-84
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• 2006

Objectives : To evaluate the neuroprotective effects of modified Bo-Yang-Hwan-O-Tang (BHT), we investigated the neuronal death protection effects to oxidative damages in N2a neuroblastoma cells. Methods : To study the cytotoxic effect of BHT on N2a neuronal cells, the cell viability was determined by MTT assay. To investigate the neuronal death protection of BHT, N2a neuronal cells were induced oxidative damages by H2O2, and assayed the cell viability and DNA fragmentation. We also investigated DPPH free radical scavenging effects of BHT by tube test. Results : In MTT assay, $500{\mu}g/ml$ of BHT was not showed cytotoxic effect on N2a neuronal cells. BHT protected N2a neuronal cells from H2O2-induced cell death in a dose-dependent manner. BHT also protected N2a neuronal cells from H2O2-induced DNA fragmentation. BHT scavenged DPPH free radicals in a dose-dependent manner. Conclusion : These data suggest that BHT may have strong anti-oxidant effects through the free radical scavenging and neuroprotective effects in neuronal cells.

• Journal of Oriental Neuropsychiatry
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• v.16 no.2
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• pp.73-88
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• 2005

Object : This study was designed to asses the effect of Kunnuetang and herbs on Mouse neuroblastoma 2a cells damaged by hypoxia-reoxygenation. Method : Mouse neuroblastoma 2a (N2a) cells were measured by MTT assay and LDH assay after 48h hypoxia and 6h reoxygenation. Mouse neuroblastoma 2a (N2a) cells were treated by Kunnuetang, Duchang and Daejo. Result : 1. Kunnuetang, was effective on LDH assay of hypoxia and reoxygenation. 2. Duchang and Daejo were generally effective on LDH assay of hypoxia and reoxygenation. 3. In MTT assay of hypoxia Kunnuetang was not effective. Duchung and Daejo were not generally effective on MTT assay, but in certain condition Herbs were effective. 4. In MTT assay of reoxygenation Kunnuetang and Daejo were not effective. But Duchung was effective in certain condition. Conclusion : The results suggest that Kunnuetang, Duchang and Daejo may have protective effect on vascular dementia and ad patient.

• Journal of Oriental Neuropsychiatry
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• v.17 no.2
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• pp.37-59
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• 2006

Objective : This study was designed to asses the effect of Hwangryunhaedoktang and herbs on Mouse neuroblastoma 2a cells damaged by hypoxia-reoxygenation. Method : Mouse neuroblastoma 2a (N2a) cells were measured by MTT assay and LDH assay after 48h hypoxia and 6h reoxygenation. Mouse neuroblastoma 2a (N2a) cells were treated by Hwangryunhaedoktang and herbs. Result : 1. Hwangryunhaedoktang was effective on LDH assay of hypoxia and reoxygenation. 2. All of herbs were generally effective on LDH assay of hypoxia and reoxygenation. In LDH assay of hypoxia, the effects of herbs depended on concentration. In MTT assay of hypoxia, Coptidis Rhizoma and Gardeniae Fructus were effective. In MTT assay of reoxygenation most of herbs were not effective. But Phellodendri Cortex was effective in high concentration. Conclusion : The results imply that Hwangryunhaedoktang and all herbs of it nay have protective effect on dementia and aging.

• Journal of Oriental Neuropsychiatry
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• v.17 no.2
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• pp.15-36
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• 2006

Objective : This study was designed to assess effect of and Chenwangbosim-Dan(CWBSD) herbs on Mouse neuroblastoma 2a cells damaged by hypoxia-reoxygenation. Method : Mouse neuroblastoma 2a (N2a) cells were measured by MTT assay and LDH assay after 48h hypoxia and 6h reoxygenation. Mouse neuroblastoma 2a (N2a) cells were treated by CWBSD and herbs. Result : 1. In MTT assay of hypoxia CWBSD and BJI, SJH, IS, CHR, HS among all of herbs were effective. Especially CWBSD and IS were highly effective. 2. In MTT assay of reoxygenation SJI, SJH, VJ, IS, BJI were effective. Especially SJI, SJH, YJ were highly effective. 3. In LDH assay of hypoxia CWBSD, DS, DG, SJH, OMZ were effective. Especially CWBSD, DG were highly effective. 4. In LDH assay of reoxygenation all of herbs except CWBSD and BJI were generally effective. Especially CHR, SJH, YJ, OMZ, HS were highly effective. Conclusion : The results suggest that CWBSD, and it's ingradient(especially SJH, CHR and SJI) may have protective effect on condition of oxidative stress.

• Journal of Oriental Neuropsychiatry
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• v.16 no.1
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• pp.19-41
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• 2005

Object : This study was designed to asses the effect of Sunghyangjungkisan-ga-pogongng and herbs on Mouse neuroblastoma 2a cells damaged by hypoxia-reoxygenation. Method : Mouse neuroblastoma 2a (N2a) cells were measured by MTT assay and LDH assay after 48h hypoxia and 6h reoxygenation, Mouse neuroblastoma 2a (N2a) cells were treated by SHJG+P and herbs. Result : 1. SHJG+P was effective on LDH assay of hypoxia and reoxygenation. 2. The herbs were generally effective on LDH assay of hypoxia and reoxygenation. In MTT assay of hypoxia JP and GC were effecctive. In LDH assay of hypoxia all of herbs were effective. DMH, BC, SY, NS were more effective than other herbs. In LDH assay of reoxygenation KH, BH, BBR, DMH were especially effective. In MTT assay of reoxygenation most of herbs were not effective. But GC, SY, BH, JP were effective. Conclusion : The results imply that SHJG+P and all of berbs may have protective effect on dementia and GC, SY, BH, JP may have protective effect.

• Archives of Pharmacal Research
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• v.27 no.8
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• pp.834-839
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• 2004

In human neuroblastoma SK-N-BE(2) cells undergoing apoptotic death induced by ginsenos-ide Rh2, a dammarane glycoside that was isolated from Panax ginseng C. A. Meyer, caspase-1 and caspase-3 were activated. The expression of Bax was increased in the cells treated with ginsenoside Rh2, whereas Bcl-2 expression was not altered. Treatment with caspase-1 inhibi-tor, Ac-YVAD-CMK, or caspase-3 inhibitor, Z-DEVD-FMK, partially inhibited ginsenoside Rh2-induced cell death but almost suppressed the cleavage of the 116 kDa PARP into a 85 kDa fragment. When the levels of p53 were examined in this process, p53 accumulated rapidly in the cells treated early with ginsenoside Rh2. These results suggest that activation of caspase-1 and -3 and the up-regulation of Bax are required in order for apoptotic death of SK-N-BE(2) cells to be induced by ginsenoside Rh2, and p53 plays an important role in the pathways to promote apoptosis.

• Advances in pediatric surgery
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• v.10 no.2
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• pp.107-111
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• 2004

Recently, the incidence of perinatally detected asymptomatic adrenal gland masses has increased because of widespread use of radiological diagnostic tools. However, optimal treatment of these masses has not been determined. The aim of this study is to elucidate the treatment guideline of perinatally diagnosed adrenal gland masses. The authors retrospectively reviewed the medical records of the 11 patients with asymptomatic adrenal gland mass, detected perinatally, between 1999 and 2004. Six cases were detected by prenatal ultrasound and 5 cases were incidentally detected by postnatal ultrasound. Six patients (surgery group) underwent mass excision. The pathologic diagnoses were neuroblastoma (n=4), adrenocortical adenoma (n=1) and adrenal pseudocyst (n=1). The indications for operation were suspicion of neuroblastoma (n=5) or absence of size decrease during observation (n=1). Three of the 5 suspicious cases of neuroblastoma and one case under observation were proven to be neuroblastoma. There was no surgical complication in the urgery group. All neuroblastoma patients have been well during the follow up period ($24.4{\pm}14.4$ month) without evidence of recurrence. Five cases (observation group) were closely observed because of the benign possibility or size decrease in follow up ultrasound. During the observation period ($39{\pm}21$ week), 4 cases showed complete spontaneous resolution and 1 case showed markedly decreased size of the mass but could not be followed up completely. Surgical resection of the perinatally diagnosed asymptomatic adrenal gland mass is a safe treatment method especially in case of suspicion of neuroblastoma, but closed observation can be applied.

• Journal of Genetic Medicine
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• v.1 no.1
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• pp.45-50
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• 1997

Neuroblastoma, a pediatric malignant neoplasm of neural crest origin, has a wide range of clinical virulence. The mechanisms contributing to the development of neuroblastomas are largely unclear, but non-random chromosomal changes identified over the past years suggest the involvement of genetic alterations. Amplification of the human N-myc proto-oncogene is frequently seen either in extrachromosomal double minutes or in homogeneously staining regions (HSRs) of aggressively growing neuroblastomas. N-myc maps to chromosome 2 band 24, but HSR have never been observed at this band, suggesting transposition of N-myc during amplification. We have constructed and analyzed the region-specific painting probe for HSR in neuroblastoma IMR-32 to determine the derivative chromosomes. Microdissection was performed on HSR using an inverted microscope with the help of microglass needles and an micromanipulator. We pretreated the microdissected fragments with Topoisomerase I which catalyzes the relaxation of supercolled DNA, and performed two initial rounds of DNA synthesis with T7 DNA polymerase followed by conventional PCR to enable the reliable preparation of Fluorescent in situ hybridization probe from a single microdissected chromosome. With this method, it was possible to construct the region-specific painting probe for HSR. The probe hybridized specifically to the HSRs of IMR-32, and to 2p24, 2p13 of normal chromosome. Our results suggest there was coamplification of N-myc together with DNA of the chromosome 2p24 and 2p13. Moreover, the fluorescent signals for the amplified chromosomal regions in IMR-32 cells were also easily recognized at a Thus this painting probe can be applied to detect the similar amplification of N-myc in neuroblastoma tissue, and the probe pool for HSR may be used to identify the cancer-relevant genes.