• 생명과학회지
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• 제21권8호
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• pp.1109-1119
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• 2011

예로부터 한국, 일본 및 중국에서 민간처방 약재 및 건강식품으로 사용되어온 매실은 다양한 효능을 가지는 것으로 알려져 있으나 인체 암세포에서 유발하는 항암작용 및 그에 따른 분자생물학적 기작에 대해서는 명확히 밝혀져 있지 않다. 본 연구에서는 인체 혈구암 U937 세포에서 매실의 메탄올 추출물(MEPM)이 유발하는 항암효과 및 항암기전을 조사한 결과, MEPM 처리 농도 의존적으로 암세포의 성장억제 및 apoptosis를 유발하는 것으로 나타났다. MEPM에 의해서 유발되는 apoptosis에는 XIAP 및 survivin 등과 같은 IAP family의 발현 감소와 더불어 FasL의 발현 증가, Bcl-2의 발현 감소 및 Bid의 단편화 현상이 관여하는 것으로 나타났으며, 두 가지 apoptosis 유발 개시 및 최종 apoptosis 단계에서 중요한 역할을 하는 caspase-8과 -9 및 -3의 활성화와 그에 따른 다양한 기질단백질의 발현 감소 및 단편화가 동반되었음을 알 수 있었다. 또한 인위적인 caspase-3의 활성 차단으로 MEPM에 의하여 유발되는 apoptosis가 현저하게 억제되는 것으로 나타났다. 이상의 결과에서 MEPM은 암세포의 chemotherapeutic agent로서의 가능성을 확인하였지만 향후 지속적인 연구를 통하여 활성물질의 동정 및 관련 기전의 비교 등이 지속적으로 이루어져야 할 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제28권1호
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• pp.115-121
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• 2018

Upon sensing of microbial infections or endogenous danger signals in macrophages, inflammasome signaling plays a significant role in triggering inflammatory responses via producing interleukin (IL)-$1{\beta}$. Recent studies revealed that active caspase-1, a product of the inflammasome complex, causes maturation of inactive pro-IL-$1{\beta}$ into the active form. However, the underlying mechanism by which this leaderless cytokine is secreted into the extracellular space remains to be elucidated. In this study, we demonstrated that prolonged lipopolysaccharide (LPS) treatment to macrophages could trigger the unexpected maturation and extracellular release of IL-$1{\beta}$ through a nucleotide-binding oligomerization domain-like receptor family, pyrin domain-containing 3 (NLRP3)-independent manner. Short-term treatment (less than 6 h) of LPS induced robust production of the IL-$1{\beta}$ precursor form inside cells but did not promote the maturation and secretion of IL-$1{\beta}$ in bone marrow-derived macrophages or peritoneal macrophages. Instead, prolonged LPS treatment (more than 12 h) led to a significant release of matured IL-$1{\beta}$ with no robust indication of caspase-1 activation. Intriguingly, this LPS-triggered secretion of IL-$1{\beta}$ was also observed in NLRP3-deficient macrophages. In addition, this unexpected IL-$1{\beta}$ release was only partially impaired by a caspase-1 and NLRP3 inflammasome inhibitor. Collectively, our results propose that prolonged exposure to LPS is able to drive the maturation and secretion of IL-$1{\beta}$ in an NLRP3 inflammasome-independent manner.

• 대한한방부인과학회지
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• 제35권3호
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• pp.1-23
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• 2022

Objectives: The purpose of this study is to evaluate anti-breast cancer and anti-inflammatory effects of Chungganhaewool-tang and Shipyeukmiyeugi-eum. Methods: MDA-MB-231 cells were used to measure cytotoxicity, Reactive oxygen species (ROS) production, protein expression amounts of Bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), B-cell lymphoma-extra large (Bcl-xl), Cytochrome C Caspase-3, Caspase-7, Caspase-9, Poly ADP-ribose polymerase (PARP), Nuclear factor erythroid-2-related factor 2 (Nrf2), Heme oxygenase-1 (HO-1) and NAD (P) H Quinone Oxidoreductase 1 (NQO1) to evaluate the anti-breast cancer effects of Chungganhaewool-tang (CHT) and Shipyeukmiyeugi-eum (SYE), and THP-1 cells, differentiated into macrophage and induced inflammation with Lipopolysaccharide (LPS), were used to measure production amounts of ROS, Nitric oxide (NO), and protein expression amounts of Inducible nitric oxide synthase (iNOS), Cyclooxygenase (COX-2), Interleukin-1 beta (IL-1β), Interleukin-6 (IL-6) and Tumor necrosis factor-alpha (TNF-α) to evaluate the anti-inflammatory effects of CHT and SYE. Results: CHT and SYE reduced MDA-MB-231 cell counts, increased protein expression of Bax and Cytochrome C, and decreased protein expression of Bcl-2, Bcl-xl. The protein expression amounts of Caspase-3, 7, and 9 decreased, but amounts of the active form, cleaved Caspase-3, 7, and 9, increased. In addition, PARP protein expression decreased, the amount of PARP protein in the cleaved form increased, and the amount of protein expressions of Nrf2 and HO-1 decreased, but NQO1 showed no significant difference. In THP-1 cells CHT and SYE reduced ROS and NO, and reduced protein expressions of iNOS, COX-2, IL-1, and TNF-α, but only SYE groups reduced IL-6. Conclusions: This study suggests that CHT and SYE have potential to be used as treatments for breast cancer.

• 생명과학회지
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• 제26권7호
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• pp.847-854
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• 2016

Nucleoside adenosine 유도체의 하나인 cordycepin (3′-deoxyadenosine)은 Cordyceps 속에서 유래된 활성 물질 중의 하나로서 항염증, 항산화 및 항암활성을 포함한 다양한 약리학적 효능이 있는 것으로 잘 알려져 있다. 본 연구에서는 AGS 인체 위암세포의 증식에 미치는 cordycepin의 영향과 관련 기전 연구를 시도하였다. Cordycepin의 처리에 따라 AGS 세포의 생존율이 처리 농도 의존적으로 감소되었으며, DNA 단편화 및 flow cytometery 분석에 따른 apoptosis 유발 또한 유의적으로 증가하였음을 확인하였다. 이러한 cordycepin 처리에 따른 AGS 세포의 apoptosis 유도에는 TRAIL, DR5 및 FasL의 mRNA 및 단백질의 발현 증가가 연관되어 있었다. 아울러 cordycepin은 Bcl-2 family 중 pro-apoptotic 인자인 Bax의 발현은 증가시켰으며, anti-apoptotic 인자인 Bcl-2 및 Bcl-xL의 발현은 전사 및 번역 수준에서 억제시켰다. 이러한 현상들은 extrinsic 및 intrinsic apoptosis의 initiator caspase (caspase-8 및 -9) 뿐만 아니라 effector caspase인 caspase-3의 활성과 PARP 단백질의 절단 증가와 연관성이 있었다. 따라서 AGS 세포에서 cordycepin에 의한 apoptosis의 유발은 death receptor 활성과 mitochondria 기능 손상을 포함한 multiple apoptotic pathway가 관여할 것으로 생각된다. 비록 좀 더 세심한 기전 연구의 결과가 뒤따라야 되겠지만, 본 연구의 결과는 cordycepin의 항암작용을 이해하는데 중요한 자료가 될 것이며 향후 수행될 추가 실험을 위한 기초 자료로서 그 가치가 매우 높을 것으로 생각된다.

• 대한한방부인과학회지
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• 제27권2호
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• pp.12-22
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• 2014

Objectives: This study aimed to evaluate the effects of Cuscutae Semen water extract (CS) on MCF-7 human breast cancer cells. Methods: To clarify the results, we cultivated MCF-7 cells in cell culture plates. And then we extracted each of $100{\mu}g/ml$, $300{\mu}g/ml$, $600{\mu}g/ml$ CS, gave it to MCF-7 cell. After these process we performed MTT assay to elucidate the ability of apoptosis. The result of mRNA was analyzed by RT-PCR. Results: Each of concentrated extracts CS decreased the survival rate of MCF-7 cells. CS decreased Bcl-2 which is known as a blocking cell apoptosis. Bax, caspase-3, P21 and RIP-1 that accelerate apoptogenic activity factors increased by CS. CS did not change the condition of caspase-8, caspase-9, P53 factors on MCF-7 cells. Furthermore caspase-8, caspase-9, P53 factors on MCF-7 cells does not make it more active but turn it on. Conclusions: According to the above results, we could suggest that CS can occur the apoptosis on MCF-7 cells.

• 동의생리병리학회지
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• 제22권3호
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• pp.630-641
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• 2008

Gamisamgibopae-tang (GMSGBPT) is a traditional Korean medicine, which has been used for patients suffering from a lung disease in Oriental medicine. In the present study, we examined the biochemical mechanisms of apoptosis by GMSGBPT in NCI-H460 and A549 human non-small-cell lung cancer cell lines. It was found that GMSGBPT could inhibit the cell proliferation of A549 cells in a concentration-dependent manner, however GMSGBPT did not affect the cell proliferation of NCI-H460 cells. Apoptotic cell death in A549 cells were detected using DAPI staining and annexin V fluorescein methods. The induction of apoptotic cell death by GMSGBPT was connected with a down-regulation of anti-apoptotic Bcl-2 and Bcl-xL expression, and proteolytic activation of caspase-3 and caspase-9 in A549 cells. However, GMSGBPT did not affect the levels of pro-apoptotic Bax and Bad expression, and activity of caspase-8. GMSGBPT treatment also concomitant degradation and/or inhibition of poly (ADP-ribose) polymerase (PARP), ${\beta}$-catenin, phospholipase C-1 (PLC${\gamma}$1) and DNA fragmentation factor 45/inhibitor of caspase-activated DNase (DFF45/ICAD). Taken together, these findings suggest that GMSGBPT may be a potential chemotherapeutic agent for the control of human non-small-cell lung cancer cells and further studies will be needed to identify the active compounds that confer the anti-cancer activity of GMSGBPT.

• Archives of Pharmacal Research
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• 제28권9호
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• pp.1047-1052
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• 2005

Two phenolic glucosides, eutigoside Band eutigoside C were isolated from the fresh leaves of Eurya emarginata. These two phenolic glucosides exerted a significant inhibitory effect on the growth of HL-60 promyelocytic leukemia cells. Furthermore, when the HL-60 cells were treated with eutigoside C, several apoptotic characteristics such as DNA fragmentation, morphologic changes, and increase of the population of sub-G1 hypodiploid cells were observed. In order to understand the mechanism of apoptosis induction by eutigoside C, we examined the changes of Bcl-2 and Bax expression levels. The eutigoside C reduced BcI-2 protein and mRNA levels, but slightly increased Bax protein and mRNA levels in a time-dependent manner. When we examined the activation of caspase-3, an effector of apoptosis, the eutigoside C increased the expression of active form (19-kDa) of caspase-3 and the increase of their activities was demonstrated by the cleavage of poly (ADP-ribose) polymerase, a substrate of caspase-3, to 85-kDa. The results suggest that the inhibitory effect of eutigoside C from E. emarginata on the growth of HL-60 appears to arise from the induction of apoptosis via the down-regulation of BcI-2 and the activation of caspase.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9153-9157
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• 2014

Background: Capparis spinosa L., a Uygur medicine, had been shown to have anti-tumor activity in our early experiments with an N-butanol extract (CSBE) as its active fraction. However, the mechanisms responsible for its effects are not clearly understood. Here, we report that treatment of SGC-7901 cells with CSBE resulted in dose-dependent reduction of cell viability and induction of apoptosis. Materials and Methods: To observe the inhibitory and killing effects of CSBE on SGC-7901, the SRB method was adopted, apoptosis being observed by electron microscopy. To clarify the mechanisms of apoptosis, Western blot and enzyme-labeled methods were used to examine the release of cytochrome c (Cyt c) and the activation of the caspase cascade. Results: By electron microscopy, apoptotic morphologic changes were detectable after CSBE administration. In this study, it was also demonstrated that CSBE induced apoptosis in SGC-7901 cells by inhibiting mPTP open, mitochondrial cytochrome c release, caspase-9 and caspase-3 activation. Conclusions: The findings indicated that CSBE induces aap optosis through mitochondrial pathway.

• Asian Pacific Journal of Cancer Prevention
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• 제15권15호
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• pp.6321-6326
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• 2014

${\alpha}$-Methyl-n-butylshikonin (MBS), one of the active components in the root extracts of Lithospermum erythrorhizon, posses antitumor activity. In this study, we assess the molecular mechanisms of MBS in causing apoptosis of SW620 cells. MBS reduced the cell viability of SW620 cells in a dose-and time-dependent manner and induced cell apoptosis. Treatment of SW620 cells with MBS down-regulated the expression of Bcl-2 and up-regulated the expression of Bak and caused the loss of mitochondrial membrane potential. Additionally, MBS treatment led to activation of caspase-9, caspase-8 and caspase-3, and cleavage of PARP, which was abolished by pretreatment with the pan-caspase inhibitor Z-VAD-FMK. MBS also induced significant elevation in the phosphorylation of JNK and p38. Pretreatment of SW620 cells with specific inhibitors of JNK (SP600125) and p38 (SB203580) abrogated MBS-induced apoptosis. Our results demonstrated that MBS inhibited growth of colorectal cancer SW620 cells by inducing JNK and p38 signaling pathway, and provided a clue for preclinical and clinical evaluation of MBS for colorectal cancer therapy.

• Journal of Microbiology and Biotechnology
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• 제15권3호
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• pp.532-536
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• 2005

In the course of our screening for small molecules to inhibit apoptosis of U937 human leukemia cells induced by etoposide ($10\;{\mu}g/ml$), Penicillium sp. F020150 with potent inhibitory activity was selected. The active compound was purified from ethyl acetate extract of the microorganism by Sephadex LH-20 column chromatography and HPLC, and was identified as gentisyl alcohol (2,5-dihydroxybenzyl alcohol) by spectroscopic methods. The compound inhibited caspase-3 induction with $IC_{50}$ value of $3.0\;{\mu}g/ml$ after 8 h of etoposide treatment. The expression levels of caspase-3 and PARP were dose-dependently inhibited by the compound, suggesting that gentisyl alcohol inhibits etoposide-induced apoptosis via downregulation of caspases.