• 대한의생명과학회지
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• 제15권2호
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• pp.147-152
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• 2009

Lycopus lucid us Turcz is well known as traditional Chinese medicine, and it has been shown to exhibit antiinflammatory, -allergic and -oxidative effect. However, its anti-cancer properties have not been examined yet. In this study, we investigated the effect of the methanol extract of Lycopus lucid us Turcz on anti-cancer effect in MCF-7 human breast cancer cells. Treatment of Lycopus lucidus Turcz extract induced apoptosis and inhibition of cell proliferation in dose- and time-dependent manner. Apoptosis in the MCF-7 cells was characterized with the changes in nuclear morphology; decrease of Bcl-2 and caspase-7 expression; and increase of cleaved poly ADP-ribose polymerase(PARP). Furthermore, treatment of Lycopus lucidus Turcz extract caused the down-regulation of cell cycle-related protein including, cdk4, cyclin D1 and E2F-1. These results suggest that Lycopus lucidus Turcz might have the therapeutic value against human breast cancer cells.

• 약학회지
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• 제56권1호
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• pp.35-41
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• 2012

In this study, we examined the effect of luteolin to enhance TRAIL-induced anticancer effect in SK-Hep1 cells. We found that combined use of TRAIL with luteolin markedly enhanced the cytotoxicity compared to either agent alone by inducing apoptosis. Furthermore, combined treatment of TRAIL with luteolin significantly induced activation of death receptor pathway-related proteins as well as PARP-cleavage and activation of effector caspases. Also, our result indicated that upregulation of DR4 and DR5 by luteolin combination may contribute to enhanced susceptibility of SK-Hep1 cells to TRAIL.

• 생명과학회지
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• 제25권11호
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• pp.1255-1264
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• 2015

천궁(C. officinale)은 예로부터 민간처방 약재로 사용되었으며, 항염증, 항산화, 항암 및 신생혈관억제 등의 효능을 가지는 것으로 알려져 있다. 하지만 혈구암세포에서 apoptosis 유발과 관련된 분자생물학적 기전에 대해서는 명확히 밝혀져 있지 않다. 본 연구에서는 인체 혈구암세포인 U937 세포에서 천궁의 열수, 에탄올 및 메탄올 추출물(WECO, EECO 및 MECO)이 유발하는 항암효과 및 항암기전을 조사하였다. 먼저 WECO, EECO 및 MECO가 유발하는 증식억제 정도를 조사한 결과 EECO가 가장 뛰어난 효능을 가진다는 것을 알 수 있었으며, 이러한 현상이 apoptosis 유발에 의한 것임을 annexin-V 염색, apoptotic body 형성, DNA 단편화 및 MMP 소실 등을 통하여 확인하였다. EECO 처리에 의한 apoptosis 유발에는 DR4의 발현 증가와 함께 cIAP-1, Bcl-2 및 total Bid의 발현감소가 관여하였으며, caspases-3, -8 및 -9의 활성화와 함께 caspases-3의 기질 단백질인 PARP, β-catenin 및 PLC γ1의 단편화도 관찰되었다. 또한 EECO는 AMPK signaling pathway를 활성화시키는 것으로 나타났으며, AMPK 억제제인 compound C를 이용하여 AMPK의 활성을 억제하였을 경우 EECO에 의하여 유발되었던 apoptosis가 현저하게 감소되는 것으로 나타났다. 이상의 결과를 살펴볼 때 인체 혈구암세포인 U937 세포에서 EECO에 의하여 유발되는 apoptosis는 AMPK가 중요한 조절자로서 작용하는 것으로 생각된다.

• 한국식품영양과학회지
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• 제35권4호
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• pp.395-401
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• 2006

본 연구에서는 해조류 중 홍조류에 속하는 참가사리를 추출, 분획하여 항발암효과를 측정하였다. 참가사리 분획물을 4종의 암세포주 HT29, HepG2, MCF-7 및 H16-F10에 처리하였을 때 암세포 증식 억제실험을 한 결과 대장암세포주인 HT29의 경우 GTMM층과 GTMB층에서 농도의존적인 효과가 나타났으며 첨가농도 $150{\mu}g/mL$에서 각각 93.64%와 74.14%의 수치를 보였다. 간암세포주인 HepG2의 경우 $150{\mu}g/mL$ 첨가시 GTMM층과 GTMB층이 각각 95.97%, 81.78%의 높은 증식 억제효과를 보였으며 유방암세포주인 MCF-7에서는 GTMM층이 $120 {\mu}g/mL$과 $150{\mu}g/mL$ 첨가농도에서 각각 88.50%와 91.82%의 높은 암세포증식 억제효과를 나타냈다. 피부암세포주인 B16-F10은 다른 세포주에 비해 미약하나 역시 GTMM층이 최종첨가농도에서 86.20%의 억제를 나타내었다. 이와 같이 4종의 암세포주의 증식억제는 전반적으로 GTMM층과 GTMB층에서 높은 효과가 나타남을 알 수 있다. 한편 Western blot analysis를 통해 간암세포 주인 HepG2에서 GTMM층의 처리에 따른 pro-apoptosis Bcl-2 family의 발현증가를 볼 수 있었으며, PARP 분절과 연관된 caspase-3, 7의 활성화를 확인할 수 있었다. 이처럼 GTMM층은 apoptosis의 작용에 의한 세포사멸을 유도하며, 이러한 apoptosis의 기전으로는 최소한 caspase의 활성화에 따른 절단 PARP 단백질의 증가, Bad, Bax 등의 apoptosis 관여인자가 작용하고 있음을 확인할 수 있었다. HepG2를 이용한 quinone reductase 유도활성여부를 측정한 결과 GTMM층이 $45{\mu}g/mL$ 및 $60{\mu}g/mL$의 시료첨가농도에서 대조군에 비해 각각 2.34, 2.86배의 QR 유도활성을 나타내었으며, GTMB층의 경우 최종첨가농도인 $60{\mu}g/mL$에서 2.04배의 효소활성을 나타내었다.

• The Korean Journal of Physiology and Pharmacology
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• 제13권4호
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• pp.281-285
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• 2009

Rotenone, a mitochondrial complex I inhibitor, can induce the pathological features of Parkinson's disease (PD). In the present study, naringin, a grapefruit flavonoid, inhibited rotenone-induced cell death in human neuroblastoma SH-SY5Y cells. We assessed cell death and apoptosis by measuring mitogen-activated protein kinase (MAPKs) and caspase (CASPs) activities and by performing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Naringin also blocked rotenone-induced phosphorylation of Jun NH2-terminal protein kinase (JNK) and P38, and prevented changes in B-cell CLL/lymphoma 2 (BCL2) and BCL2-associated X protein (BAX) expression levels. In addition, naringin reduced the enzyme activity of caspase 3 and cleavages of caspase 9, poly (ADP-ribose) polymerase (PARP), and caspase 3. These results suggest that naringin has a neuroprotective effect on rotenone-induced cell death in human neuroblastoma SH-SY5Y cells.

• 생명과학회지
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• 제26권4호
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• pp.398-405
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• 2016

3-Hydroxy-3-methylglutaryl coenzyme A 환원효소의 억제제로 알려진 statin은 고지혈증 치료제로 널리 사용되고 있고, 또한 다양한 암에서 항암효과를 나타낸다고 알려져 있다. 최근 연구에서는 reactive oxygen species (ROS)가 세포사멸 신호에 중요한 역할을 한다고 보고하였지만, rosuvastatin에 의한 ROS 생성의 정확한 기전은 아직 밝혀지지 않았다. 인간 전립선암 세포주인 PC-3 세포를 이용하여 rosuvastatin에 의한 세포사멸 경로를 확인하였다. 세포독성, 세포사멸과 ROS의 생성을 측정하기 위해서 MTT assay, annexin V/PI 염색과 DCFH-DA염색을 통해 flow cytometry에 의해 측정하였다. 본 연구의 결과에서, rosuvastatin은 농도와 시간 의존적으로 세포 생존율 감소와 세포형태변화를 확인할 수 있었다. Flow cytometry 분석을 통해 세포주기와 apoptosis를 확인하였고 Western blotting assay를 통하여 PARP와 procaspase-3가 감소되는 것을 통해 apoptosis를 재확인 할 수 있었다. 또한 rosuvastatin은 농도 의존적으로 ROS 생산을 증가하였고, ROS 생성 저해제인 N-acetylcysteine (NAC) 처리를 통해 ROS와 apoptosis가 회복되었다. 따라서 rosuvastatin이 ROS 생성을 통해 apoptosis를 유도한다는 것을 알 수 있었고, 이는 인간 전립선 암세포에 대한 항암치료제로서의 가능성을 제시한다.

• 대한한방내과학회지
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• 제30권4호
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• pp.845-857
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• 2009

Background and Objectives : Korean mistletoe lectin (Viscum album coloratum agglutinin, VCA) and bee venom (BV) have been reported to induce apoptosis in various cancer cell lines in vitro and to show antitumor activity against a variety of tumors in animal models. However, the comparative effect of VCA and BV on the anti-cancer effect and mechanisms of action has not been determined. In this study, the effect in a human hepatocellular carcinoma cell line, Hep G2 cells, was examined. Methods : Cytotoxic effects of VCA and BV on Hep G2 cells were determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay in litro. The apoptotic cell death was then confirmed by propidium iodide staining and DNA fragmentation analysis. The mechanisms of action were examined by the expression of anti-apoptotic proteins and activation of mitogen-activated protein kinases. The involvement of kinase was examined in VCA or BV-induced apoptosis by using kinase inhibitors. Results : VCA and BV killed Hep G2 cells in a time and dose-dependent manner. Treatment of Hep G2 cells with VCA activated poly (ADP-ribose) polymerase-1 (PARP-1) known as a marker of apoptosis, and mitogen-activated protein kinases signaling pathways including MAPK/ERK, p38 MAPK and JNK. BV also activated PARP-1, MAPK/ERK. and p38 MAPK but not JNK. The expression level of anti-apoptotic molecule, Bcl-X, was decreased by VCA treatment but not by BV. Finally, the phosphorylation level of ERM proteins involved in the cytoskeleton homeostasis was decreased by both stimuli. VCA-induced apoptosis was partially inhibited by in the presence of JNK and p38 inhibitor, but BV only by p38 inhibitor. Conclusions : VCA-induced apoptosis is dependent on the activation of p38 and JNK. while BV-induced apoptosis is mediated by p38 activation in Hep G2 cells.

• 대한한방내과학회지
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• 제33권3호
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• pp.272-283
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• 2012

Objectives : The purpose of this study was to investigate the mechanism of protective effect of Jinmu-tang (JMT, Zhenwu-tang) extract on $H_2O_2$-induced cell death in C6 glial cells. Methods : Cultured C6 glial cells of white mice were pretreated with JMT extract and exposed to $H_2O_2$ for inducing cell death. We measure the cell viability by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and investigate the cell morphology using a light microscope after crystal violet (CV) staining. Reactive oxygen species (ROS) formation was analyzed using a flow cytometer and a fluorescent microscope after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). DNA fragmentation was analyzed using a flow cytometer after propidium iodide (PI) staining and nuclei morphology was investigated using a fluorescent microscope after 2-[4-amidinophenyl]-6-indo-lecarbamidine dihydrochloride (DAPI) staining. We analyzed expression of Bax, processing of procaspase-3 and poly (ADP-ribose) polymerase (PARP), and activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) by western blot method. Tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) secretion was analyzed using Quantikine kit. Results : We determined the elevated cell viability by JMT extract on $H_2O_2$-induced C6 glial cell death. ROS formation, DNA fragmentation, $I{\kappa}B{\alpha}$ phosphorylation, NF-${\kappa}B$ activation, and secretion of TNF-${\alpha}$ induced by $H_2O_2$ are inhibited by JMT extract pre-treatment. JMT extract inhibits Bax expression, processing of caspase-3 and PARP that are critical biochemical markers of apoptotic cell death. Conclusions : These results suggest that JMT extract has a protective effect on $H_2O_2$-induced C6 glial cell death in various pathways.

• 생명과학회지
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• 제19권9호
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• pp.1209-1217
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• 2009

많은 암세포에 선택적 세포독성을 나타내는 TNF-related apoptosis-inducing ligand (TRAIL)는 유효한 항암제로서 사용될 수 있지만, TRAIL에 내성을 나타내는 암세포에는 TRAIL-sensitization의 방법이 필요하다. 사람 대장암 세포인 KMl2 세포도 TRAIL에 내성을 나타낸다. 본 연구에서는 KMl2세포의 TRAIL 내성에 대한 새로운 표적 분자의 발굴과 이를 토대로 한 새로운 내성극복 방법을 연구하였다. 새로운 TRAIL sensitizer로서 quercetin을 발굴하고, 이를 KMl2세포에 TRAIL과 병용 처리하여 TRAIL의 효과증강을 시도하였다. KMl2세포에서 quercetin은 c-FLTP의 발현을 감소시키고, DNA-PK/Akt 신호전달경로를 억제하므로서, death receptors (DR4/DR5) 발현을 증강시켰다. 또한 caspases (caspase 3, -8 및 -9)활성 증강과 PARP cleavage, 이에 따른 Bax의 발현을 증강시키는 기전으로 TRAIL에 의한 apoptosis를 증대시키는 활성이 있음을 밝혔다. 즉 quercetin이 KMl2세포의 TRAIL-sensitization에 사용될 수 있음을 제시하였다. 이러한 연구 결과는 대장암의 치료 시 TRAIL과 quercetin병용하므로서 치료 효과를 높일 수 있는 새로운 약제 병용 방법을 제시하였고, 다른 TRAIL 내성 종양에 응용 될 수 있음을 시사하였다.

• 동의신경정신과학회지
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• 제24권4호
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• pp.419-434
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• 2013

Objectives: The object of this study is to observe the cognition and motor function recovery effects of Joojakwhan (JJW), a traditional Korean poly-herbal formula for treating various neuropsychiatric diseases such as dementia, for the mildly stroke rats, with 60 minutes of reperfusion transient middle cerebral artery occlusion (tMCAO). Methods: In the present study, 125, 250 and 500 mg/kg of JJW were orally administered, once per day for 10 continuous days 2 hours after the tMCAO. The body weight changes, infarct sizes under 2% 2, 3, 5-triphenyl tetrazolium chloride (TTC) stain, sensorimotor functions and cognitive motor behavior tests were serially monitored with cerebral caspase-3 and cleaved poly (ADP-ribose) polymerase (PARP)-immunoreactivities and histopathological changes. The effects of tMCAO on sensorimotor functions were evaluated by using of limb placing and body-swing tests, and the cognitive motor behaviors were also observed with water maze tests. Results: From the results of tMCAO, with marked decreases of body weights, disorders of sensorimotor functions increases the limb placing test scores, and decrease the numbers and percentages of body swings to the ipsilateral sides. The cognitive motor behaviors increases the distances and time to reach the escape platform which included the inhibitions of the decreases with repeated trials that were observed with focal cerebral cortex infarct volumes. In addition, the marked increases of the atrophy, numbers of degeneration, caspase-3- and PARP-immunoreactive cells around peri-infarct ipsilateral cerebral cortex were also observed in tMCAO controls when compared with the sham control rats, respectively. Conclusions: The results obtained from this study suggest that oral administrations of JJW indicate obvious cognitions and motor function recoveries of the rats with tMCAO, mild strokes, which are mediated by neuro-protective effects through known antioxidant effects of components.