• 대한한방내과학회지
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• 제35권1호
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• pp.79-91
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• 2014

Objectives : To investigate the anti-cancer effect of Palbohoichoon-tang (PBHCT) extracts. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were microscopically analyzed after staining with $10{\mu}M$ 2-[4-amidinophenyl]-6-indolecarbamidine dihydrochloride (DAPI) and TUNEL. We also analyzed expression of Bcl2, $Bcl_{xL}$, Bax, procaspase-3, procaspase-9, and procyclic acidic repetitive protein (PARP) by western blot method. Results : Observations showed that PBHCT induced the apoptotic cell death proved by increased sub-G1 phase cell population, apoptotic body formation and chromatin condensation. Western blot analysis of total cell lysates revealed that the PBHCT induced cleavage of caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP). In addition, PBHCT dose-dependently increased the activity of caspase-9, caspase-3 and PARP-1. Furthermore, PBHCT reduced anti-apoptotic Bcl2, $Bcl_{xL}$ expression which contributed to the loss of mitochondrial membrane potential and the activations of caspase-9 and caspase-3. Conclusions : These findings suggest that PBHCT exerts anti-cancer effects on human neuroblastoma SH-SY5Y cells by inducing apoptotic death via down-regulation of anti-apoptotic proteins such as Bcl2 and $Bcl_{xL}$, up-regulation of pro-apoptotic proteins such as Bax, and activation of caspase cascades and PARP-1.

• 동의생리병리학회지
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• 제17권5호
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• pp.1309-1314
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• 2003

We examined the effects of Oak Smoke Flavoring (OSF, Holyessing) on the cell proliferation of DU145 and PC3 human prostate carcinoma cell line. OSF treatment resulted in a concentration-dependent inhibition of the cell viability in both DU145 and PC3 cell lines. The anti-proliferative effects by OSF treatment in DU145 and PC3 cells were associated with morphological changes such as membrane shrinking and cell rounding up. DNA flow cytometric histograms showed that population of S phase of the cell cycle was increased by OSF treatment in a dose-dependent manner. Western blot analysis revealed that cyclin B1 and cdc2 proteins were reduced by OSF treatment in DU145 cells, whereas cyclin A was markedly inhibited in PC3 cells. Furthermore, we observed an increase of Cdk inhibitor p16 and p27 protein, and an inhibition of phosphorylation of pRB by OSF treatment in a dose-dependent manner. The present results indicated that OSF-induced inhibition of human prostate carcinoma cell proliferation is associated with the blockage of S phase progression.

• Asian Pacific Journal of Cancer Prevention
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• 제17권12호
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• pp.5189-5193
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• 2016

Objective: Dorema glabrum Fisch. & C.A. Mey is a perennial plant that has several curative properties. Anti-proliferative activity of seeds of this plant has been demonstrated in a mouse fibrosarcoma cell line. The aim of the present study was to evaluate cytotoxicity of D. glabrum root extracts in a human gastric adenocarcinoma (AGS) cell line and explore mechanisms of apoptosis induction, cell cycle arrest and altered gene expression in cancer cells. Materials and Methods: The MTT assay was used to evaluate IC50 values, EB/AO staining to analyze the mode of cell death, and flow cytometry to assess the cell cycle. Quantitative real-time polymerase chain reaction (qRT-PCR) amplification was performed with apoptosis and cell cycle-related gene primers, for cyclin D1, c-myc, survivin, VEGF, Bcl-2, Bax, and caspase-3 to determine alteration of gene expression. Results: Our results showed that n-hexane and chloroform extracts had greatest toxic effects on gastric cancer cells with IC50 values of $6.4{\mu}g/ml$ and $4.6{\mu}g/ml$, respectively, after 72 h. Cell cycle analysis revealed that the population of treated cells in the G1 phase was increased in comparison to controls. Cellular morphological changes indicated induction of apoptosis. In addition, mRNA expression levels of Bax and caspase-3 were increased, and of bcl-2 survivin, VEGF, c-myc and cyclin D1 were decreased. Conclusion: Our study results suggest that D. glabrum has cytotoxic effects on AGS cells, characterized by enhanced apoptosis, reduced cell viability and arrest of cell cycling.

• Asian Pacific Journal of Cancer Prevention
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• 제16권10호
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• pp.4357-4361
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• 2015

Thunbergia Laurifolia Linn. (TL) is one of the most familiar plants in Thai traditional medicine that is used to treat various conditions, including cancer. However, the antitumor activity of TL or its constituents has never been reported at the molecular level to support the folklore claim. The present study was designed to investigate the antitumor effect of an aqueous extract of TL in human breast cancer cells and the possible mechanism(s) of action. An aqueous crude extract was prepared from dried leaves of TL. Folin-Ciocalteu colorimetric assays were used to determine the total phenolic content. Antiproliferative and cell cycle effects were evaluated in human breast adenocarcinoma MCF-7 cells by MTT reduction assay, cell growth inhibition, clonogenic cell survival, and flow cytometric analysis. Free radical generation by the extracts was detected using electron paramagnetic resonance spectroscopy. The exposure of human breast adenocarcinoma MCF-7 cells to a TL aqueous extract resulted in decreases in cell growth, clonogenic cell survival, and cell viability in a concentration-dependent manner with an $IC_{50}$ value of $843{\mu}g/ml$. Treatments with extract for 24h at $250{\mu}g/ml$ or higher induced cell cycle arrest as indicated by a significant increase of cell population in the G1 phase and a significant decrease in the S phase of the cell cycle. The capability of the aqueous extract to generate radical intermediates was observed at both high pH and near-neutral pH conditions. The findings suggest the antitumor bioactivities of TL against selected breast cancer cells may be due to induction of a G1 cell cycle arrest. Cytotoxicity and cell cycle perturbation that are associated with a high concentration of the extract could be in part explained by the total phenolic contents in the extract and the capacity to generate radical intermediates to modulate cellular proliferative signals.

• 대한한방내과학회지
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• 제28권2호
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• pp.230-241
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• 2007

Objective : This study was intended to ascertain the protective effect of phenolic-rich fraction (PRF) from Fructus Schisandrae on SH-SY5Y cells. Methods : PRF was obtained from the 80% ethanol extract of Fructus Schisandrae by Sepabeads SP-850 column chromatography. The neuroprotective effect of the FS PRS was investigated due to the hydrogen peroxide $(H_2O_2)-induced$ apoptosis of cultured SH-SY5Y cells. Results : Cell viability assays revealed that pretreating SH-SY5Y cells with PRF (10-200 ${\mu}g/mL$) resulted in significant dose-dependent protection against $H_2O_2-induced$ cell death. The effect was assessed by flow cytometric analysis of DNA contents using propidium iodide (PI) staining. The population of apoptotic cells was increased by 32.89% in only $H_2O_2$ (150 ${\mu}M$)-treated environment, but it was reduced by pre-treatment of FS PRF (200 ${\mu}g/mL$) to 21.61%. $H_2O_2-induced$ caspase-3 activation and PARP cleavage were reduced in FS PRF pre-treated cells, and PRF led to an apparent suppressive effect on the oxidative stress induced by reactive oxygen species (ROS). Conculsion : This study showed that Fructus Schisandrae should be useful for the treatment prevention of neurodegenerative diseases associated with elevated ROS levels.

• 한국환경생태학회지
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• 제26권3호
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• pp.322-332
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• 2012

본 연구는 원격무선추적 자료를 바탕으로 우리나라의 농촌지역에 서식하는 삵의 행동권, 서식지 선택, 개체군 위협요인을 파악하고자 하였으며 3개체(수컷 2, 암컷 1)를 대상으로 한 연구결과는 다음과 같다. 첫째, 행동권은 평균 $2.64{\pm}1.99km^2$(K 95)와 $3.69{\pm}1.34km^2$(MCP 100)이었으며, 핵심공간은 평균 $0.64{\pm}0.47km^2$(K 50)를 나타냈고, 겨울철에 추적된 수컷의 행동권이 가장 넓었다($5.19km^2$, MCP 100). 둘째, Jacobs index에 기반을 둔 Johnson의 서식지 선택모형을 분석한 결과 삵은 2단계 선택(경관 내에서의 서식지 선택)에서 초지와 논을 선호하고 산림은 회피하는 반면에, 3단계 선택(행동권 내에서의 서식지 선택)에서는 하천변의 초지를 매우 선호하고 논을 회피하였다. 셋째, 로드킬은 삵의 심각한 개체군 위협요인으로 판단되며, 논의 토지피복 비율이 높고, 하천변에 도로가 존재하며, 산림이 마을 주위에만 소규모로 존재하는 지역에서는 삵의 개체군 유지가 매우 불안정할 수 있어 이러한 지역에 로드킬 대책과 하천변 초지보호 등의 노력이 보다 집중되어야 할 것이다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권24호
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• pp.10659-10663
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• 2015

Background: Rapamycin is an effective anti-angiogenic drug. However, the mode of its action remains unclear. Therefore, in this study, we aimed to elucidate the antitumor mechanism of rapamycin, hypothetically via apoptotic promotion, using MCF-7 breast cancer cells. Materials and Methods: MCF-7 cells were plated at a density of $1{\times}10^5$ cells/well in 6-well plates. After 24h, cells were treated with a series of concentrations of rapamycin while only adding DMEM medium with PEG for the control regiment and grown at $37^{\circ}C$, 5% $CO_2$ and 95% air for 72h. Trypan blue was used to determine the cell viability and proliferation. Untreated and rapamycin-treated MCF-7 cells were also examined for morphological changes with an inverted-phase contrast microscope. Alteration in cell morphology was ascertained, along with a stage in the cell cycle and proliferation. In addition, cytotoxicity testing was performed using normal mouse breast mammary pads. Results: Our results clearly showed that rapamycin exhibited inhibitory activity on MCF-7 cell lines. The $IC_{50}$ value of rapamycin on the MCF-7 cells was determined as $0.4{\mu}g/ml$ (p<0.05). Direct observation by inverted microscopy demonstrated that the MCF-7 cells treated with rapamycin showed characteristic features of apoptosis including cell shrinkage, vascularization and autophagy. Cells underwent early apoptosis up to 24% after 72h. Analysis of the cell cycle showed an increase in the G0G1 phase cell population and a corresponding decrease in the S and G2M phase populations, from 81.5% to 91.3% and 17.3% to 7.9%, respectively. Conclusions: This study demonstrated that rapamycin may potentially act as an anti-cancer agent via the inhibition of growth with some morphological changes of the MCF-7 cancer cells, arrest cell cycle progression at G0/G1 phase and induction of apoptosis in late stage of apoptosis. Further studies are needed to further characterize the mode of action of rapamycin as an anti-cancer agent.

• Tuberculosis and Respiratory Diseases
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• 제53권5호
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• pp.485-496
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• 2002

연구 방법 : Nonsteroidal anti -inflammatory drugs (NSAIDs)는 대장앙의 항암 예방약제로 사용되고 었다 지속적으로 NSAIDs를 복용하면 대장암에 걸릴 위험도가 40-50% 감소되는 것으로 알려져 있다. NSAIDs가 대장암에서 종양의 크기를 감소시키는 것에 대한 정확한 기전은 알려져 있지 않으나, 일부 연구자들은 NSAIDs를 고농도로 투여하였을 때 세포 주기를 조절하는 유전자 발현의 변형과 세포고사의 유도로 설명하고 있다. 그러나 폐암에서 NSAIDs의 암 예방효과에 대해 확립 된 바 없어, 저자들은 NCI-H1299 세포주에서 NSAIDs가 세포고사를 유도하는지 알아보고자 하였다. 방 법 : 세포 독성은 MTT 방법으로 측정하였고, 세포고사를 알아보기 위해 유세포 분석과 핵산 염색을 시행하였다. 세포고사의 기전을 알아보기 위해 caspase family의 활성을 측정하였고, 세포고사의 마지막 단계인 PARP와 ICAD의 분절을 westem blot으로 확인하였다. 결 과 : NCI-H1299 세포에서 NaSaL 처리 시 생존율이 농도와 시간에 의존적으로 유의하게 감소하였고, 생존율의 감소는 세포주기에서 $subG_0/G_1$의 증가와 핵산 염색시 핵의 분절의 관찰로서 세포고사가 일어남을 관찰하였다. 10 mM NaSaL 처리 후 caspase-3 protease의 활성은 24시간에 증가하여 30시간에 최고에 이르고 감소하였으나 caspase-6, 8, 9 proteases의 활성은 의미 있는 증가가 없었다. PARP와 ICAD의 분절은 농도와 시간 의존적으로 증가하였다. 결 론 : NCI-H1299 폐암 세포주에서 NaSaL은 caspase-3 protease의 활성을 통하여 유도되었다.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1988년도 학술대회지
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• pp.47-54
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• 1988

세포막의 주요 구성성분인 콜레스테롤은 고등생명체의 성장 및 생존에 필수적인 물질이다. 그러나 혈중 콜레스테롤의 농도가 너무 높으면 동맥경화증을 유발하여 매우 치명적이다. 본 연구는 고려인삼(Panax ginseng C.A. Meyer) 뿌리의 주요성분의 하나인 사포닌류의 고콜레스테롤 혈중 억제효과를 규명하기 위해 시도되었다. 고콜레스테롤 식이를 인삼사포닌과 함께 또는 단독으로 투여한 토끼나 쥐에 $^{125}I-LDL$을 주입한 후 혈액으로부터 $^{125}I-LDL$의 제거속도를 추적한 결과 고콜레스테롤 식이만을 투여한 대조군보다 인삼사포닌을 함께 투여한 시험군이 $^{125}I-LDL$의 제거속도가 훨씬 빨랐다. 고콜레스테롤식이를 투여한 쥐의 LDL수용체 생합성에 미치는 ginsenoside 혼합물과 정제된 ginsenoside $Rb_{1},\;Rb_{2},\;Re,\;Rg_{1}$의 영향을 조사하였다. 쥐 간의 LDL 수용체 수준을 분석한 결과 시험군이 대조군에 비해 크게 높았으며 다른 장기(신장, 정소, 부신피질)의 경우에도 간에서의 결과와 유사하였다. 또한 간파쇄액을 효소원으로 사용하여 콜레스테롤 부터의 담즙산 합성에 미치는 ginsenoside의 영향을 시험관내에서 관찰한 결과 반응 혼합물에서의 ginsenoside의 농도가 $(10^{-3}-10^{-4}\%)$일 때 담즙산합성이 현저히 증가되었다. 위와 같은 실험결과로 미루어 보아 ginsenoside는 세포 내부에서의 콜레스테롤 대사를 촉진하여 세포내의 콜레스테롤 농도를 저하시킴으로써 콜레스테롤의 LDL 수용체 합성 억제를 완화시켜주는 것이라고 생각된다.성분들은 cyclooxygenase에 직접 작용하지 않는다는 것을 설명해 준다. 2. Panaxadiol (500 ${\mu}g/ml$은 $PGE_{2}$ 생성에는 영향이 없으나 $PGF_{2}{\alpha}$생성에 유의적인 차이를 보이지 않으나 농도의존적으로 $TxB_{2}$의 생성을 감소시켰고 6-keto-$PGF_{1}{\alpha}$의 생성을 증가시켰는데 이는 $TxA_{2}$ synthetase 억제제인 imidazole의 효과와 유사하였다. 4. G-Re는 $1{\times}10^{-5}g/ml$ 이하의 농도에서는 효과가 없으나 $1{\times}10^{-4}g/ml$ 이상의 농도에서 농도의존적으로 유의성 있는 $PGE_{2},\;PGF_{2}{\alpha},\;TXB_{2}$의 생성억제와 함께 6-keto-$PGF_{1}{\alpha}$ 증가를 보였다. 이는 prostacyclin synthetase를 자극하는 serotonin의 효과와 같은 작용으로서 prostacyclin synthetase 억제제인 tranylcypromine에 대하여 길항효과를 보였다. 5. $TxB_{2}$생성억제 작용을 나타내는 ginsenoside들의 효과를 뒷받침하기 위하여 인삼 saponin 성분을 전처치한 patelet rich plasma에서 혈소판 응집시험 결과, ADP로 유도된 혈소판 응집반응에는 모든 인삼 saponin 성분들이 효과가 없었으나 arachidonic acid로 유도된 혈소판 응집반응에는 $G

• 생명과학회지
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• 제29권3호
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• pp.295-302
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• 2019

Liquiritigenin (LG)은 licorice 뿌리에서 분리된 chiral flavonoid이다. LG는 항산화, 항암 및 항염증 효과를 포함하여 다양한 생물학적 활성을 가지고 있다. 구강편평세포암종에서 LG의 항암 활성은 아직 밝혀지지 않았다. 본 연구에서는 구강편평상피암 세포(HN22)에서 LG의 항암 효능을 시험하였다. HN22 세포에 LG를 처리하여 MTT 분석으로 세포 생존율을 평가하였으며, Annecin V/7-Aminactinomycin D 염색, 세포주기 및 Multi-caspase 활성을 $Muse^{TM}$ cell Analyzer로 분석하여 세포사멸 유도를 확인하였다. 분석결과, 구강편평상피암 HN22 세포에 LG를 처리시 G2/M 세포주기 정지를 일으켰으며, Western blotting 통하여 cyclin B1 및 CDC2 발현 감소와 p27 발현 증가를 확인하였다. LG는 활성산소종의 생성을 유발하고, CCAAT/enhancer-binding protein homologous protein (CHOP) 및 78-kDa glucose regulated protein (GRP78)의 발현을 농도의존적으로 유도하였다. HN22 세포에 LG의 처리는 미토콘드리아 막전위의 손실(${\Delta}{\Psi}m$)을 일으켰다. LG를 처리한 HN22 세포의 단백질 분석결과 apoptotic protease activating factor-1 (Apaf-1), cleaved Poly (ADP-Ribose) Polymerase (C-PARP) 및 Bax의 발현을 증가함을 확인하였다. 따라서 우리의 결과는 LG이 구강편평상피암 세포의 세포사멸을 유도하여 항암제 역할을 할 수 있는 천연 화합물임을 시사한다.