• 한국식품과학회지
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• 제34권1호
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• pp.114-117
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• 2002

연교의 메탄올 추출물로부터 caspase 저해물질을 분리하였으며, ESI-MS, $^1H-NMR$, $^13C-NMR$, DEPT 등의 기기분석 자료에 의하여 rengyolone으로 동정하였다. 이 물질은 $IC_{50}\;6.25\;{\mu}g/mL$의 농도에서 etoposide가 처리된 U937 세포주의 caspase-3 유도 저해를 나타내었다. 또한 rengyolone은 $Interleukin-1{\beta}$가 처리된 D10S 세포에서 caspase-1의 유도저해활성을 나타내었으며, $IC_{50}$값은 $7.5\;{\mu}g/mL$이었다.

• 통합자연과학논문집
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• 제7권3호
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• pp.166-172
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• 2014

Caspases, a family of cysteinyl aspartate-specific proteases plays a central role in the regulation and the execution of apoptotic cell death. Activation of caspases-3 stimulates a signaling pathway that ultimately leads to the death of the cell. Hence, caspase-3 has been proven to be an effective target for reducing the amount of cellular and tissue damage. In this work, comparative molecular field analysis (CoMFA) was performed on a series of 3, 4-dihydropyrimidoindolones derivatives which are inhibitors of caspase-3. The best predictions were obtained for CoMFA model ($q^2=0.676$, $r^2=0.990$). The predictive ability of test set ($r^2_{pred}$) was 0.688. Statistical parameters from the generated QSAR models indicated the data is well fitted and have high predictive ability. Our theoretical results could be useful to design novel and more potent caspase-3 derivatives.

• 통합자연과학논문집
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• 제7권4호
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• pp.227-233
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• 2014

Caspases, a family of cysteinyl aspartate-specific proteases plays a central role in the regulation and the execution of apoptotic cell death. Activation of caspases-3 stimulates a signaling pathway that ultimately leads to the death of the cell. Hence, caspase-3 has been proven to be an effective target for reducing the amount of cellular and tissue damage. In this work, comparative molecular similarity indices analysis (CoMSIA) was performed on a series of 3,4-dihydropyrimidoindolones derivatives which are inhibitors of caspase-3. The best predictions were obtained for CoMSIA model ($q^2$ = 0.586, $r^2$ = 0.955). The predictive ability of test set ($r^2_{pred}$) was 0.723. Statistical parameters from the generated QSAR models indicated the data is well fitted and have high predictive ability. Our theoretical results could be useful to design novel and more potent caspase-3 derivatives.

• Natural Product Sciences
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• 제22권4호
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• pp.293-298
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• 2016

Plant-derived triterpenoids commonly possesses biological properties such as anti-inflammatory, antimicrobial, anti-viral and anti-cancer. Luvunga scandens is one of the plant that produced triterpenoids. The aims of the study was to analyze cell cycle profile and to determine the expression of p53 unregulated modulator of apoptosis (PUMA), caspase-8 and caspase-9 genes at mRNA level in MCF-7 cell line treated with two triterpenoids, flindissol (1) and 3-oxotirucalla-7,24-dien-21-oic-acid (2) isolated from L. scandens. The compounds were tested for cell cycle analysis using flow cytometer and mRNA expression level using quantitative RT-PCR. The number of MCF-7 cells population which distributed in Sub G1 phase after treated with compound 1 and 2 were 7.7 and 9.3% respectively. The evaluation of the expression of genes showed that both compounds exhibited high level of expression of PUMA, caspase-8 and caspase-9 as normalized to ${\beta}-actin$ via activation of those genes. In summary, the isolated compounds of L. scandens plant showed promising anticancer properties in MCF-7 cell lines.

• Molecules and Cells
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• 제24권1호
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• pp.95-104
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• 2007

The mechanism of acacetin-induced apoptosis of human breast cancer MCF-7 cells was investigated. Acacetin caused 50% growth inhibition ($IC_{50}$) of MCF-7 cells at $26.4{\pm}0.7{\mu}M$ over 24 h in the MTT assay. Apoptosis was characterized by DNA fragmentation and an increase of sub-G1 cells and involved activation of caspase-7 and PARP (poly-ADP-ribose polymerase). Maximum caspase 7 activity was observed with $100{\mu}M$ acacetin for 24 h. Caspase 8 and 9 activation cascades mediated the activation of caspase 7. Acacetin caused a reduction of Bcl-2 expression leading to an increase of the Bax:Bcl-2 ratio. It also caused a loss of mitochondrial membrane potential that induced release of cytochrome c and apoptosis inducing factor (AIF) into the cytoplasm, enhancing ROS generation and subsequently resulting in apoptosis. Pretreatment of cells with N-acetylcysteine (NAC) reduced ROS generation and cell growth inhibition, and pretreatment with NAC or a caspase 8 inhibitor (Z-IETD-FMK) inhibited the acacetin-induced loss of mitochondrial membrane potential and release of cytochrome c and AIF. Stress-activated protein kinase/c-Jun $NH_4$-terminal kinase 1/2 (SAPK/JNK1/2) and c-Jun were activated by acacetin but extracellular-regulated kinase 1/2 (Erk1/2) nor p38 mitogen-activated protein kinase (MAPK) were not. Our results show that acacetin-induced apoptosis of MCF-7 cells is mediated by caspase activation cascades, ROS generation, mitochondria-mediated cell death signaling and the SAPK/JNK1/2-c-Jun signaling pathway, activated by acacetin-induced ROS generation.

• Journal of Microbiology and Biotechnology
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• 제32권8호
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• pp.1034-1040
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• 2022

Fas-associated death domain (FADD) is an adapter molecule that bridges the interaction between receptor-interacting protein 1 (RIP1) and aspartate-specific cysteine protease-8 (caspase-8). As the primary mediator of apoptotic cell death, caspase-8 has two N-terminal death-effector domains (DEDs) and it interacts with other proteins in the DED subfamily through several conserved residues. In the tumor necrosis receptor-1 (TNFR-1)-dependent signaling pathway, apoptosis is triggered by the caspase-8/FADD complex by stimulating receptor internalization. However, the molecular mechanism of complex formation by the DED proteins remains poorly understood. Here, we found that direct DED-DED interaction between FADD and caspase-8 and the structure-based mutations (Y8D/I128A, E12A/I128A, E12R/I128A, K39A/I128A, K39D/I128A, F122A/I128A, and L123A/I128A) of caspase-8 disrupted formation of the stable DED complex with FADD. Moreover, the monomeric crystal structure of the caspase-8 DEDs (F122A/I128A) was solved at 1.7 Å. This study will provide new insight into the interaction mechanism and structural characteristics between FADD and caspase-8 DED subfamily proteins.

• Journal of Life Science
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• 제9권1호
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• pp.58-63
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• 1999

Highly metastatic mouse T-lymphoma CS21 cells can grow in vitro when cocultured with CA12 lymph node stromal cells, but they undergo apoptotic cell death when separated from CA12 stromal cells. It has been found that cysteine and interleukin-7(IL-7) as antiapoptotic soluble factors that produced by CA12 stromal cells. In this study, we report that an ICE family protease is activated in CS21 cells when separated from CA12 stromal cells and cultured alone. Enzyme purification using an avidin affinity column revealed that the involved cysteine protease possessed caspase3-like death protease activity. In addition, when IL-7 was added to CS21 cell culture, the protease activity could not be detected during partial purification of the enzyme. Taken together, these results strongly suggest that the caspase3-like protease activation is suppressed by IL-7 as an antiapoptotic factor that leads to abrogation of apoptosis execution.

• 생명과학회지
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• 제18권9호
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• pp.1177-1185
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• 2008

폴리아민은 미생물에서부터 동.식물에 이르기 까지 모든 세포들에서 발견되는 극성을 띈 분자이다. 세포의 성장과 분화에 폴리아민이 중요한 역할을 한다는 것은 이미 오래 전부터 알려져 온 사실이나 정확한 작용 기작은 잘 밝혀져 있지 않다. 최근에 와서는 폴리아민이 다양한 방법으로 세포 독성을 유발한다는 결과가 보고되고 있다. 본 연구에서는 spm의 세포독성 효과가 세포 내 칼슘이온 농도 증가에 따른 미토콘드리아-의존 기작에 의하여 일어난다는 것을 보여준다. Spm에 의해 유도된 세포 내 칼슘이온 농도 증가는 주로 외부로부터의 칼슘 유입에 의한 것으로 생각되며, 세포 내 칼슘 증가는 미토콘드리아로부터의 cytochrome c 방출과 미토콘드리아막의 탈분극에 의한 membrane potential 변화를 초래하였다. 세포사멸에 주도적인 역할을 하는 caspase의 확인에 있어서는, MCF-7 세포는 caspase-3이 결핍되어서 caspase-7이 중심적인 역할을 하는 것으로 이미 알려져 있다. 본 연구에서 확인 한 결과 spm 처리 시 caspase-7과 -12가 활성화되었다. 또한 세포사멸 조절 단백질인 Bcl-2 종류 단백질들의 발현을 조사 한 결과 세포사멸 억제 단백질인 Bcl-2의 발현은 크게 억제되었으며, 촉진단백질인 Bax는 spm 처리시 단백질 양이 거의 2배로 증가되었다. 이상의 결과에 의하면, spm에 의해 유도되는 세포사멸과정은 세포질 내 칼슘이온 농도 증가에 의한 미토콘드리아의 변화가 주도적인 역할을 하는 것으로 생각된다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권7호
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• pp.3223-3228
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• 2016

Reports indicate that 15-deoxy-delta-12,14-prostaglandin-J2 (15d-PGJ2) has anticancer activities, but its mechanisms of action have yet to be fully elucidated. We therefore investigated the effects of 15d-PGJ2 on the human breast cancer cell lines, MCF-7 (estrogen receptor $ER{\alpha}+/ER{\beta}+$) and MDA-MB-231 ($ER{\alpha}-/ER{\beta}+$). Cellular proliferation and cytotoxicity were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays while apoptosis was determined by fluorescence microscopy and flow cytometry using annexin V-propidium iodide (PI) staining. ER expression was determined by Western blotting. Intracellular calcium was stained with Fluo-4 AM while intracellular caspase activities were detected with Caspase-$FLICA(R)$ and measured by flow cytometry. We showed that 15d-PGJ2 caused a significant increase in apoptosis in MCF-7 and MDA-MB-231 cells. $ER{\alpha}$ protein expression was reduced in treated MCF-7 cells but pre-incubation with the $ER{\alpha}$ inhibitor' ICI 182 780' did not affect the percentage of apoptotic cells. The expression of $ER{\beta}$ was unchanged in both cell lines. In addition, 15d-PGJ2 increased intracellular calcium ($Ca^{2+}$) staining and caspase 8, 9 and 3/7 activities. We therefore conclude that 15d-PGJ2 induces caspase-dependent apoptosis that is associated with an influx of intracellular $Ca^{2+}$ with no involvement of ER signaling.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1053-1062
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• 2007

유방암 치료제로 사용되는 탐옥시펜 (Tam)은 MCF-7 세포주에서 세포증식을 억제하며 세포사멸을 유도한다. 본 연구에서는 Tam의 세포독성 효과가 세포 내 칼슘이온 농도 증가에 따른 미토콘드리아-의존 기작에 의하여 일어난다는 것을 보여준다. Tam에 의해 유도된 세포 내 칼슘이온 농도 증가는 주로 외부로부터의 칼슘 유입에 의한 것으로 생각된다. 칼슘 채널 억제제를 이용한 실험 결과에 의하며, 칼슘 증가 초기 단계는 주로 전압의존 칼슘채널에 의한 것이며 후기에는 세포 내 저장된 칼슘의 유출, 혹은 다른 방법에 의한 칼슘 유입으로 생각된다. Tam에 의한 세포 내 칼슘 증가는 미토콘드리아로부터의 cytochrome c 방출과 미토콘드리아막의 탈분극에 의한 membrane potential 변화를 초래하였다. 세포사멸에 주도적인 역할을 하는 caspase의 확인에 있어서는, MCF-7 세포는 caspase-3이 결핍되어서 caspase-7이 중심적인 역할을 하는 것으로 이미 알려져 있다. 본 연구에서 확인한 결과 Tam 처리시 caspase-7이 활성화되었으며, 또한 세포사멸 조절 단백질인 Bcl-2 종류 단백질들의 발현을 조사 한 결과 세포사멸 억제 단백질인 Bcl-2의 발현에는 변화가 없었으나 촉진단백질인 Bax는 Tam 처리시 단백질 양이 2배로 증가되었다. 이상의 결과에 의하면, Tam에 의해 유도되는 세포사멸과정은 세포질 내 칼슘이온 농도증가에 의한 미토콘드리아의 변화가 주도적인 역할을 하는 것으로 생각된다.