• Journal of Gastric Cancer
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• 제3권2호
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• pp.75-79
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• 2003

Purpose: Evidence exists that dysregulation of apoptosis is involved in the pathogenesis of cancer development. The Bcl-$x_{L}$/Bcl-2-associated death promoter (BAD), a member of the Bcl-2 family, is a critical regulatory component of the intrinsic cell-death pathway that exerts its pro-apoptotic effect upon heterodimerization with anti-apoptotic proteins Bcl-2 and Bcl-$X_{L}$. Expression of the BAD protein has been reported in several cancer types, but not in stomach cancer. The aim of this study was to explore the expression status of the BAD protein in gastric carcinomas. Materials and Methods: In the current study, we analyzed the expression of the BAD protein in 60 advanced gastric adenocarcinomas by using immunohistochemistry and a tissue microarray approach. Results: Immunopositivity (defined as $\geq\30\%$) was observed for the BAD protein in 57 ($95\%$) of the 60 cancers. Normal gastric mucosal cells showed weaker expressions of the BAD protein than gastric carcinomas. Conclusion: Taken together, these results suggest that stomach cancer cells in vivo may need BAD protein expression for apoptosis. Also, the higher expression of the BAD protein in stomach cancer cells than in normal gastric mucosal cells suggests that apoptosis might be easily triggered in susceptible stomach cancer cells, thereby producing selective pressure to make more apoptosis-resistant cells during tumor development.

• Natural Product Sciences
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• 제23권4호
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• pp.227-234
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• 2017

Moringa oleifera Lam (M. oleifera, Moringaceae) is a tree of the Moringaceae family that can reach a height of between 5 and 10 m. The current paper presents cytotoxic effect of M. oleifera fruits and its flavonoids 1 and 2. The viability of HCT116 human colon cancer cells were 38.5% reduced by $150{\mu}g/mL$ of ethanolic extracts in a concentration-dependent manner; in addition, we observed the apoptotic features of cell shrinkage and decreased cell size. Bcl-2 family proteins were regulated as determined by Western blotting analysis, suggesting that M. oleifera fruits and their flavonoids 1 and 2 induced apoptosis through an intrinsic pathway. Based on our findings, 70% ethanolic extracts of M. oleifera fruits and flavonoids 1 and 2 might be useful as cytotoxic agents in colorectal cancer therapy.

• BMB Reports
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• 제39권5호
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• pp.556-559
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• 2006

The Bcl-2 family of proteins regulates mitochondrial functions during cell death by modulating the efflux of death-promoting proteins such as cytochrome c and endonuclease G. Upon the binding of death ligands to their receptors, caspase-8 cleaves Bid, a BH3-only protein, into tBid that causes the mitochondrial damages resulting in the release of cytochrome c and endonuclease G. Also, another BH3-only protein, hNoxa, has been shown to induce the efflux of cytochrome c from the mitochondria. Whether the efflux proteins from the mitochondria in response to tBid or hNoxa are the same or different, however, has not been addressed. We have demonstrated that endonuclease G activities are not detectable among the proteins released from isolated mitochondria by hNoxa but are detectable in that by tBid. These results suggest that the efflux of proteins from the mitochondria are differentially modulated by tBid and hNoxa.

• Natural Product Sciences
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• 제26권3호
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• pp.191-199
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• 2020

Fucoidan, a natural component of brown seaweed, has various biological activities such as anti-cancer activity, anti-oxidant, and anti-inflammatory against various cancer cells. However, the fucoidan has been implicated in melanoma cells via apoptosis signaling pathway. Therefore, we investigated apoptosis with fucoidan in A2058 human melanoma cells with dose- and time-dependent manners. In our results, A2058 cells viability decreased at relatively short-time and low-concentration through fucoidan. This effects of fucoidan on A2058 cells appeared to be mediated by the induction of apoptosis, as manifested by morphological changes through DNA-binding dye Hoechst 33342 staining. When a dose of 80 ㎍/mL fucoidan was treated, the cells were observed: crescent or ring-like structure, chromatin condensation, and nuclear fragmentation. With the increase at 100 ㎍/mL fucoidan, the cell membrane is intact throughout the total process, including membrane blebbing and loss of membrane integrity as well as increase of sub-G1 DNA. Furthermore, to understand the exact mechanism of fucoidan-treated in A2058 cells, western blotting was performed to detect apoptosis-related protein expression. In this study, Bcl-2 family proteins can be regulated by fucoidan, suggesting that fucoidan-induced apoptosis is modulated by intrinsic pathway. Therefore, expression of Bcl-2 and Bax may result in altered permeability, activating caspase-3 and caspase-9. And the cleaved form of poly ADP-ribose polymerase was detected in fucoidan-treated A2058 cells. These results suggest that A2058 cells are highly sensitive to growth inhibition by fucoidan via apoptosis, as evidenced by activation of extracellular signal-regulated kinases/p38/Bcl-2 family signaling, as well as alteration in caspase-9 and caspase-3.

• 대한본초학회지
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• 제28권2호
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• pp.33-38
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• 2013

Objects : Apoptosis leads to the death of a cell. The mitochondrial pathway of apoptosis, a process regulated by the Bcl-2 family of proteins, plays a key role in various biological processes. The tuber of Liriope platyphylla Wang et TANG (Liliaceae), also known as Liriopis tuber, is famous in Oriental medicine owing to its tonic, antitussive, expectorant and anti-asthmatic properties. The purpose of this study was to observe the effect of the Liriopis tuber on mitochondrial-mediated apoptosis in PC-12 cells. Method : A cytotoxic test on Liriopis Tuber water extract was conducted and another MTT assay was conducted to observe the cytoprotective effect against 4-HNE 25 ${\mu}M$ that causes oxidative stress in PC-12 cells for 24 hours. In addition, in order to observe the expression of Bcl-2 and Bax protein involved with apoptosis, western blot was conducted. Results : The LT water extract had no toxicity for PC-12 cell. In the cytoprotective effect against 4-HNE, both of the group treated with 50 ${\mu}g/m{\ell}$ and 100 ${\mu}g/m{\ell}$ of LT water extract showed a significant increase in comparison with the control group. In Bax protein expression, all the experimental groups treated with LT water extract showed a decrease in comparison with the control group but had no significance. In Bcl-2 protein expression, all the experimental groups treated with LT water extract showed a significant increase in comparison with the control group. Conclusion : These results suggest that LT is effective in reducing apoptosis.

• 한국식품영양과학회지
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• 제37권1호
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• pp.8-15
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• 2008

목향은 국화과에 속한 다년생 식물인 Saussurea lappa의 뿌리로서 한의학에서는 구토, 설사 및 염증치료 등에 사용되고 있다. 목향 추출물 및 그 성분들은 항균 작용, 항염증 작용, 혈관생성 억제 효능 등을 지니고 있으며 위암과 대장암의 세포증식을 억제하는 것으로 알려져 있다. 그러나 현재까지 전립선암에 대한 목향의 효과는 연구된 바 없다. 본 연구에서는 목향이 전립선 암세포에 미치는 영향을 조사하기 위해 인간의 전립선에서 유래한 암세포인 LNCaP 세포의 증식과 apoptosis에 미치는 영향을 조사하였다. 목향 헥산추출물을 LNCaP 세포 배양액에 여러 농도($0{\sim}4$ mg/L)로 첨가하여 세포의 증식에 미치는 영향을 조사하였다. 목향 헥산추출물의 농도가 증가할수록 LNCaP 세포의 증식은 현저하게 감소하였고 apoptosis는 증가함을 관찰하였다. 목향 헥산추출물이 LNCaP 세포의 apoptosis 일으키는 기전을 연구하기 위하여 목향 헥산추출물을 첨가하고 세포를 48시간 배양한 후 cell lysate를 얻어 Western blot을 실시하였다. Apoptosis 과정에 작용하는 중요한 단백질 중 하나인 Bcl-2 family 단백질 중 pro-apoptotic Bcl-2 단백질인 Bak와 BH3 only Bcl-2 단백질인 truncated-Bid의 단백질 수준은 목향 헥산 추출물에 의해 유의적으로 증가한 반면 anti-apoptotic Bcl-2 단백질인 Bcl-2, Bcl-xL 및 Mcl-1 단백질 수준은 변하지 않았다. 또한 apoptosis를 집행하는 caspase의 활성 형태인 cleaved caspase-8, -9, -7, -3의 단백질 수준이 목향 헥산추출물의 처리에 의해 증가하였고 caspase-3의 표적 단백질 중 하나인 PARP의 불활성 형태인 cleaved PARP의 단백질 수준도 현저하게 증가하였다. 이 결과들은 목향 헥산 추출물이 LNCaP 세포의 apoptosis를 유도함으로써 전립선 암세포의 증식을 억제함을 보여주는 것이며 목향 헥산추출물에 의한 apoptosis 유도는 caspase 활성 증가와 Bak 및 t-Bid 단백질의 증가에 의한 것임을 제시한다. 따라서 앞으로 항암효과를 나타내는 성분의 동정 및 동물실험을 통하여 좀 더 면밀한 기전 연구가 수행된다면 목향 헥산추출물은 화학적 암예방 물질이나 치료제로 개발될 수 있을 것으로 사료된다.

• Biomolecules & Therapeutics
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• 제27권1호
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• pp.41-47
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• 2019

The apoptotic effects of shikonin (5,8-dihydroxy-2-[(1R)-1-hydroxy-4-methylpent-3-enyl]naphthalene-1,4-dione) on the human colon cancer cell line SNU-407 were investigated in this study. Shikonin showed dose-dependent cytotoxic activity against SNU-407 cells, with an estimated $IC_{50}$ value of $3{\mu}M$ after 48 h of treatment. Shikonin induced apoptosis, as evidenced by apoptotic body formation, sub-G_1$ phase cells, and DNA fragmentation. Shikonin induced apoptotic cell death by activating mitogen-activated protein kinase family members, and the apoptotic process was mediated by the activation of endoplasmic reticulum (ER) stress, leading to activation of the $PERK/elF2{\alpha}/CHOP$ apoptotic pathway, and mitochondrial $Ca^{2+}$ accumulation. Shikonin increased mitochondrial membrane depolarization and altered the levels of apoptosis-related proteins, with a decrease in B cell lymphoma (Bcl)-2 and an increase in Bcl-2-associated X protein, and subsequently, increased expression of cleaved forms of caspase-9 and -3. Taken together, we suggest that these mechanisms, including MAPK signaling and the ER- and mitochondria-mediated pathways, may underlie shikonin-induced apoptosis related to its anticancer effect.

• 한국발생생물학회지:발생과생식
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• 제12권3호
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• pp.297-303
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• 2008

본 논문은 인공적인 단백질인 MCL-1ES BH3M에 관한 것으로 MCL-1ES BH3M를 과발현시 세포사멸을 유도한다. MCL-1L을 주형으로 재조합 PCR을 통해서 MCL-1ES BH3M를 클로닝하였다. 새롭게 클로닝한 단백질인 MCL-1ES BH3M 단백질은 안정성을 유지하기 위해서 PEST 도메인이 제거되어 있으며, 다른 BCL-2 패밀리 단백질과의 결합을 조절하기 위해서 BH3도메인의 Leu-Arg-Arg-Val-Gly-Asp-Gly 서열을 7개의 Ala 잔기로 인위적으로 돌연변이를 유도하였다. MCL-1ES BH3M를 293T 세포에서 과발현할 경우 세포사멸을 유도하였고, 항-세포사멸 단백질인 MCL-1L을 같이 과발현하더라도 세포사멸을 유도하였다. 또한, 과발현시 Caspase 9과 3를 활성화하였으며 면역염색법을 통해서 MCL-1ES BH3M 과발현시 미토콘드리아에 MCL-1ES BH3M 단백질이 부분적으로 위치하는 것을 확인하였다. 이상의 결과로 MCL-1ES BH3M는 Caspase 9과 3의 활성을 통해서 세포사멸을 유도한다. 결론적으로 본 연구는 세포사멸을 유도하는 새로운 molecule을 클로닝하였고, 이 molecule에 의한 세포사멸 기능을 확인하였다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권11호
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• pp.4475-4482
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• 2014

Background: Cisplatin, a DNA damaging agent, induces apoptosis through increasing DNA fragmentation. However, identification of intrinsic resistance molecules against Cisplatin is vital to estimate the success of therapy. Bag-1 (Bcl-2-associated anthanogene) is one anti-apoptotic protein involved in drug resistance impacting on therapeutic efficiency. Elevated levels of this protein are related with increase cell proliferation rates, motility and also cancer development. For this reason, we aimed to understand the role of Bag-1 expression in Cisplatin-induced apoptosis in HeLa cervix cancer cells. Cisplatin decreased cell viability in time- and dose-dependent manner in wt and Bag-1L+HeLa cells. Although, $10{\mu}M$ Cisplatin treatment induced cell death within 24h by activating caspases in wt cells, Bag-1L stable transfection protected cells against Cisplatin treatment. To assess the potential protective role of Bag-1, we first checked the expression profile of interacting anti-apoptotic partners of Bag-1. We found that forced Bag-1L expression prevented Cisplatin-induced apoptosis through acting on Mcl-1 expression, which was reduced after Cisplatin treatment in wt HeLa cells. This mechanism was also supported by the regulation of heat shock protein (Hsp) family members, Hsp90 and Hsp40, which were involved in the regulation Bag-1 interactome including several anti-apoptotic Bcl-2 family members and c-Raf.

• 한국해양바이오학회지
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• 제1권2호
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• pp.63-70
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• 2006

본 연구에서는 U937 인체 백혈병 세포의 증식에 미치는 PTX-2의 영향을 조사한 결과, PTX-2의 처리에 따라 U937 세포는 처리 농도 및 처리 시간 의존적으로 심한 형태적 변형과 함께 증식이 억제되었다. 이러한 PTX-2 처리에 의한 U937 세포의 증식억제는 apoptosis 유발과 관련이 있었으며, 이를 DAPI staining에 의한 apoptotic body 형성, flow cytometry를 이용한 sub-G1 세포 빈도의 정량적 분석을 통하여 확인하였다. 이러한 PTX-2 처리에 의한 U937 세포의 apoptosis 유발은 Bcl-2 family에 속하는 anti-apoptotic 인자인 Bcl-$X_L$의 발현 감소 및 IAPs family에 속하는 유전자들의 선택적 발현 감소와 연관성이 있음을 알 수 있었다. 이상의 결과들은 인체 암세포에서 PTX-2의 항암작용을 이해하는데 중요한 자료가 될 것이고 나아가 PTX-2을 포함한 그와 유사한 항암제 후보물질들의 연구에 있어서 기초 자료로서 사용될 수 있을 것으로 생각된다.