• Journal of Acupuncture Research
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• 제26권3호
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• pp.49-58
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• 2009

목적 : 본 연구는 봉약침의 주요성분인 멜리틴이 전립선 암세포주인 DU-145 세포성장에 어떤 영향을 미치는지를 알아보기 위하여 시행하였다. 방법 : 멜리틴이 DU-145의 성장에 미치는 영향을 알아보기 위한 cell viability 측정으로는 WST-1 assay를, 세포자멸사의 관찰에는 DAPI(4,6-diamidino-2-phenylindole)와 TUNEL staining assay를 시행하였으며, 세포자멸사 조절단백질(calpain, Bax, caspase-3, -9, cleaved caspase-3, cleavaged PARP, cleaved caspase-9, Bcl-2, XIAP, cIAP2, Akt, p-Akt, MMP-2, MMP-13)의 관찰을 위하여 western blot analysis를 시행하였다. 결과 : 1. DU-145 세포에서 멜리틴을 처리한 후 세포자멸사가 유도되어 세포성장이 억제되었다. 2. 세포자멸사 관련 단백질 중 분리된 caspase-3, caspase-9은 유의한 증가를, Bcl-2, p-Akt, XIAP, cXIAP는 유의한 감소를 나타내었다. 결론 : 이상의 결과는 멜리틴이 인간 전립선암세포주인 DU-145의 세포자멸사를 유발함으로써 증식억제 효과가 있음을 나타낸 것으로, 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 도움이 될 것으로 기대된다.

• Journal of Acupuncture Research
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• 제22권2호
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• pp.103-110
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• 2005

Phellodendri Cortex (PC) has been used traditionally in Korea for damp heat leukorrhea with thick, yellow, discharge, foul-smelling diarrhea or dysentery. We investigated whether the Phellodendri Cortex Herbal-acupuncture Solution (PCHS) induced cell-death on SNU-17, human cervical cancer cell. 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was performed to find out the cytotoxicity of PCHS. The cell death was identified as apoptosis from the results of 4, 6-diamidineo-2-phenylindole (DAPI) staining, terminal deoxy-nucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay. The expression of proapototic gene, Bax, was increased and the expression of apoptotic gene, Caspase-3, was also increased. Considering the above results, PCHS could induce the apoptosis on SNU-17, human cervical cancer cell, via Bax-related Caspase-3 activation. And it might provide the experimental data for the clinical use of Phellodendri Cortex on cervical cancer.

• 생명과학회지
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• 제24권2호
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• pp.173-180
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• 2014

에스트로겐은 조직세포의 유형과 이들의 생리적 상태에 따라 세포증식을 촉진 또는 억제 시킬 수 있으며, 주로 에스트로겐 수용체(ER)에 의해 매개되는 신호전달 경로를 통해 작용한다. 이 연구는 특히 유방암세포에서 ER에 대한 길항제로 잘 알려진 fulvestrant (Ful)가 CHO 세포주의 증식 및 세포사멸에 미치는 영향과 이들에 대한 $17{\beta}$-estradiol (E2)의 작용에 미치는 효과를 조사하고자 하였다. 이를 위하여 먼저 다양한 농도의 E2, Ful 및 E2 plus Ful의 처리기간에 따라 세포증식에 미치는 효과를 조사하였다. Cell proliferation 분석에서, 6-10일의 처리기간 동안 E2는 1 ${\mu}M$의 농도까지는 세포증식에 영향을 주지 않았지만, 15-40 ${\mu}M$에서는 처리기간의 증가에 따라 점진적으로 현저히 세포증식을 억제하였다. 흥미롭게도 Ful 또한 1 ${\mu}M$의 농도까지는 세포증식에 영향을 주지 않았지만, 10-40 ${\mu}M$에서는 농도 및 시간 의존적인 세포증식 억제효과를 보였다. 뿐만 아니라, Ful은 10일 동안 E2와의 혼합처리에서 E2에 의한 세포증식 억제효과를 감소시키기 보다는 오히려 증대시켰다. 따라서 Ful은 CHO 세포에서 E2의 항 증식작용에 대한 길항적 효과를 갖지 않음을 제시한다. 한편, E2 및 Ful에 의한 DNA 분절효과를 확인하기 위한 TUNEL assay에서 20 ${\mu}M$의 E2로 처리된 CHO 세포주에서는 DAPI로 염색된 거의 모든 핵에서 TUNEL 양성반응이 나타났으며, 40 ${\mu}M$ Ful 및 20 ${\mu}M$ E2 plus 40 ${\mu}M$ Ful로 처리된 CHO 세포주에서는 TUNEL 양성반응을 보였으나, E2 처리군과 비교하여 현저히 낮은 비율로 나타났다. 이러한 결과는 Ful이 CHO 세포에서 E2의 항증식작용에 대한 길항적 효과를 갖지 않으며, E2와 Ful에 의한 세포증식 억제와 DNA 분절을 통한 세포사멸 관련기전은 다른 신호전달 경로를 통해 매개됨을 시사한다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.237-237
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• 2004

Parthenogenesis and culture condition are essential to intracytoplasmic sperm injection and cloning by nuclear transfer. This study investigated apoptosis and in vitro development of parthenogenetic preimplantation porcine embryos. 42∼44 h in vitro matured oocytes derived from a local abattoir were used. Apoptotic cell death was analyzed by using a terminal deoxynucleatidyl transferase mediated deoxyuridine 5-triphoshate nick-end labling (TUNEL) assay. (omitted)

• 대한한의학회지
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• 제26권4호
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• pp.87-97
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• 2005

Backgrounds: Intracerebral hemon-hage is one of the most devastating types of stroke. Ginseng radix, the root of Panax Ginseng, C. A. MEYER (Araliaceae), is one of the most famous medicinal herbs with various therapeutic applications. Objectives: In the present study, the effect of aqueous extract of Ginseng radix on intracerebral hemorrhage-induced neuronal cell death in rats was investigated. Materials and Methods: Step-down avoidance task, Nissl staining, immunohistochemistry for caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay were used for this study. Results: The present results show that hemorrhage-induced lesion volume and apoptotic neuronal cell death in the striatum were significantly suppressed by treatment with Ginseng radix, resulting in enhancement of short-ten-n memory. Conclusions: We have shown that Ginseng radix has a neuroprotective effect on stroke, and aids the recovery from central nervous system sequelae following stroke.

• 대한한의학회지
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• 제24권4호
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• pp.127-135
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• 2003

Objectives : Intracerebral hemorrhage (ICH) is one of the most devastating types of stroke. The effect of acupuncture on the intrastriatal hemorrhage-induced neuronal cell death and cell proliferation in rats is examined. Methods : Cell death and cell proliferation in rats was investigated via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and immunohistochemistry for caspase-3 and 5-bromo-2'-deoxyuridine (BrdU). Results : Results showed that apoptotic cell death in the striatum and cell proliferation in the hippocampal dentate gyrus significantly increased following intrastriatal hemorrhage in rats, and that acupunctural treatment at the Zusanli acupoint suppressed the hemorrhage-induced increase in apoptosis in the striatum and cell proliferation in the dentate gyrus. Conclusions : It is suggested that acupunctural treatment, especially at the Zusanli acupoint, may aid recovery following central nervous system sequelae following ICH.

• 한방안이비인후피부과학회지
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• 제22권2호
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• pp.50-59
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• 2009

Objectives : The aim of this study was conducted that CRE (Coptidis Rhizoma Extract) induces apoptosis in YD-10B cells, human oral squamous carcinoma cell line. Methods : In this study, YD-10B cells were exposed to CRE (0.03-0.30 mg/ml), for 6-24 hours. We measured the effects of CRE on the changes of cell viability and cell membrane, TUNEL assay of CRE-treated YD-10B cell. Results : In this study, CRE caused a decrease of viability in YD-10B cells, human oral squamous carcinoma cell line. When YD-10B cells were treated with CRE, cells showed dose-dependent manner apoptotic cell death. Conclusions : These results suggest that CRE may be potential therapeutic approach in the clinical management of oral squamous cell carcinoma.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.151-154
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• 2000

배큘로바이러스와 actinomycin D에 의해 유도되는 곤충세포 apoptosis의 누에체액에 의한 저해 효과에 대해 연구하였다. 감염 전 또는 감염 중, 배양 배지에 누에체액을 첨가함으로써 배큘로바이러스에 의해 감염된 숙주 세포의 생존율을 높은 수준으로 유지시켜 주었다. 누에체액은 또한, RNA 합성 저해제인 actinomycin D에 의해 유도된 apoptosis 역시 저해시켰다. 이러한 효과들은 TUNEL assay와 flow cytometry로 확인하였다. 추가적으로, 누에체액은 감염 후 세포 내 유전자의 발현에는 영향을 미치지 않으나 바이러스 유전자의 발현을 증진시킴을 알 수 있었다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권4호
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• pp.1617-1620
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• 2012

Background: Gold nanoparticles have recently been investigated with respect to biocompatibility according to their interactions with cells. The purpose of this study was to examine cytotoxicity and apoptosis induction by well-characterized gold nanoparticles in human breast epithelial MCF-7 cells. Methods: Apoptosis was assessed by TUNEL, cytotoxicity by MTT assay and caspase 3, 9, p53, Bax and Bcl expression by real-time PCR assays. Results: Gold nanoparticles at up to $200\;{\mu}g/mL$ for 24 hours exerted concentration-dependent cytotoxicity and significant upregulation of mRNA expression of p53, bax, caspase-3 & caspase-9, whereas expression of antiapoptotic bcl-2 was down-regulated. Conclusion: To the best of our knowledge this is the first report showing that gold nanoparticles induce apoptosis in MCF-7cells via p53, bax/bcl-2 and caspase pathways.

• Biomolecules & Therapeutics
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• 제13권4호
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• pp.207-213
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• 2005

It has been proposed that reactive oxygen species (ROS), mainly superoxide anion ($O_2^-$) and hydrogen peroxide ($H_2O_2$), may mediate oxidative stress. Production of $H_2O_2$ during oxidative phosphorylation, inflammation, and ischemia can cause oxidative stress leading to cell death. Although glucose oxidase (GOX) in the presence of glucose continuously generates $H_2O_2$, it is not clear whether GOX produces apoptotic cell death in C6 glial cells. Thus, we investigated the mechanism by which GOX induces cell death. Cells were incubated with different concentration of GOX in the presence of glucose where cell viability, TUNEL and DNA ladder were analyzed. Results indicated that GOX exhibited cytotoxicity in a dose dependent manner by MTT assay. TUNEL positive cell and DNA laddering showed that GOX-induced cytotoxicity was due to apoptosis. Western blot analysis also showed that the cleaved caspase-3 level was detected in the GOX-treated cells at 10 mU/ml and increased dramatically at 30 mU/ml. Cleaved PARP also appeared at 10 mU/ml and lasted at 20 or 30 mU/ml of GOX. Cytochrome c level was increased by GOX dose dependently, which was contrast to Bcl-2 expression level. These results suggest that GOX induces apoptosis through caspase-3 activation, which followed by cytochrome c release from mitochondria through regulating of Bcl-2 level.