• 대한본초학회지
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• 제31권1호
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• pp.77-82
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• 2016

Objectives : The study was designed to observe the effect of Polygalae Radix Preparata Cum Glycyrrhizae Radix on 4-Hydroxynonenal (4-HNE)-induced apoptosis in PC-12 cell.Methods : A cytotoxic test on Polygalae Radix Preparata Cum Glycyrrhizae Radix (PG) was conducted and another MTT assay was conducted to observe the cytoprotective effect against 4-HNE that cause oxidative stress. In addition, in order to observe the expression of Bax, Bcl-2, Caspase-3 and TNF-α protein involved with apoptosis, western blot was conducted.Results : The groups treated with 25 ㎍, 50 ㎍ and 100 ㎍ of PG water extract had no toxicity for PC-12 cell. The groups treated with 25 ㎍, 50 ㎍ and 100 ㎍ of PG water extract showed a significant increase of cell survival rate in comparison with the control group injected by only 4-HNE. The groups treated with 25 ㎍ and 50 ㎍ of PG water extract showed a significant supression on increase of Bax protein expression in the control group. The group treated with 100 ㎍ of PG water extract showed a significant promotion on decrease of Bcl-2 protein expression in the control group. The group treated with 50 ㎍ of PG water extract showed a significant supression on increase of Caspase-3 protein expression in the control group. The group treated with 25 ㎍ of PG water extract showed a significant supression on increase of TNF-α protein expression in the control group.Conclusions : These results suggest that Polygalae Radix Preparata Cum Glycyrrhizae Radix is effective in reducing apoptosis by 4-HNE-dameged cell.

• Asian Pacific Journal of Cancer Prevention
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• 제14권10호
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• pp.5855-5860
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• 2013

Phytochemicals are among the natural chemopreventive agents with most potential for delaying, blocking or reversing the initiation and promotional events of carcinogenesis. They therefore offer cancer treatment strategies to reduce cancer related death. One such promising chemopreventive agent which has attracted considerable attention is sulforaphane (SFN), which exhibits anti-cancer, anti-diabetic, and anti-microbial properties. The present study was undertaken to assess effect of SFN alone and in combination with a chemotherapeutic agent, gemcitabine, on the proliferative potential of MCF-7 cells by cell viability assay and authenticated the results by nuclear morphological examination. Further we analyzed the modulation of expression of Bcl-2 and COX-2 on treatment of these cells with SFN by RT-PCR. SFN showed cytotoxic effects on MCF-7 cells in a dose- and time-dependent manner via an apoptotic mode of cell death. In addition, a combinational treatment of SFN and gemcitabine on MCF-7 cells resulted in growth inhibition in a synergistic manner with a combination index (CI)<1. Notably, SFN was found to significantly downregulate the expression of Bcl-2, an anti-apoptotic gene, and COX-2, a gene involved in inflammation, in a time-dependent manner. These results indicate that SFN induces apoptosis and anti-inflammatory effects on MCF-7 cells via downregulation of Bcl-2 and COX-2 respectively. The combination of SFN and gemcitabine may potentiate the efficacy of gemcitabine and minimize the toxicity to normal cells. Taken together, SFN may be a potent anti-cancer agent for breast cancer treatment.

• 대한두경부종양학회지
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• 제18권2호
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• pp.142-149
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• 2002

본 연구에서는 FRTL-5 갑상선세포에서 Protein phosphatase type 1(PP-1)과 type 2A(PP-2A) 효소의 억제제인 okadaic acid에 의한 apoptosis 유도 기전에 대하여 이해하고자 하였다. FRTL-5 갑상선세포에 다양한 농도($10{\sim}80nmol$)의 okadaic acid를 처리 한 후 caspase 3와 $PLC-{\gamma}1$ 분절을 확인 한 결과 40nmol 농도에서부터 caspase 3활성과 $PLC-{\gamma}1$ 분절이 나타남을 확인하였다. Okadaic acid에 의한 apoptosis 유도 기전을 조사한 결과 anti-apoptotic 기능이 있는 Bcl-2 단백질 발현은 미약하게 감소하였으나 Bcl-xL은 농도 의존적으로 급격한 감소를 보였다. Caspase 3 효소활성을 저해하는 IAP 단백질 중 cIAP2는 okadaic acid에 영향을 받지 않았으나 cIAP1과 XIAP 단백질 발현은 농도 의존적으로 감소하였다. Okadaic acid에 의한 apoptosis 유도는 caspase 3 의존적인 기전을 보였다. Caspase 3 특이억제제를 okadaic acid와 동시에 처리 시 apoptosis가 억제되었으며 $PLC-{\gamma}1$ 단백질의 절단 현상도 방지되었다. Caspase 3의 활성을 유도하는 cytochrome c의 유리는 okadaic acid처리 농도에 의존적으로 유리하였다. 결론적으로, okadaic acid에 의한 apoptosis 유도는 caspase 3 의존적이며, Bcl-2와 IAP family 단백질 감소현상에 의하여 야기되는 것으로 생각된다.

• Radiation Oncology Journal
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• 제24권2호
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• pp.128-137
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• 2006

목적: 방사선 감수성이 다양한 동계(syngeneic) 마우스 종양들을 대상으로 50% 종양억제선량과 종양성장지연 등 방사선 감수성을 대변하는 지표와 방사선에 의해 유도되는 아포토시스 간에 상관관계가 있는지 여부를 알아보고자 하였다. 또한 아포토시스와 관련된 여러 유전물질의 기본(constitutive) 발현수준을 측정한 후 이들 상호 간의 상관관계를 분석하여 방사선 감수성을 예측할 수 있는 생물학적 표지자를 알아보고자 하였다. 대상 및 방법: 동계 마우스 종양으로는 난소암 OCa-1, 유방암 MCa-K, 편평상피세포암 SCC-VII, 섬유육종 FSa-II, 간암 HCa-1을 사용하였고 이들은 PCR-SSCP 검사상 p53이 모두 자연형인 종양들이었고 주령 $8{\sim}10$ 주인 C3H/HeJ 웅성 마우스를 사용하였다. 50% 종양억제선량과 종양성장지연 및 방사선에 의해 유도되는 아포토시스를 측정하여 이들과 방사선에 의해 유도되는 아포토시스간의 상관관계를 분석하여 방사선에 의해 유도되는 아포토시스로 방사선 감수성을 예측할 수 있는지 여부를 알아보고, 또한 아포토시스와 관련된 유전물질 $053,\;p21^{WAF1/CIP1},\;BAX,\;Bcl-2,\;Bcl-x_L,\;Bcl-X_s,\;p34$ 등의 기본 발현양상 및 발현수준을 Western blot과 농도계측기로 측정한 후 이들 상호 간의 상관관계를 분석하였다. 결과: 방사선에 의해 유도된 아포토시스의 정도와 종양성장지연과의 사이에는 통계적으로 유의한 상관관계가 존재하였다(R=0.922, p=0.026). 50% 종양억제선량과의 사이에는 통계적 유의성은 변연수준이었으나(p=0.070) 상관관계의 경향을 보였다(R=-0.848). $p21^{WAF1/CIP1}$과 p34의 기본 발현수준과 50% 종양억제선량(R=0.893, p=0.041와 R=0.904, p=0.035) 및 종양성장지연(R=-0.922, p=0.026와 R=-0.890, p=0.043) 사이에는 통계적으로 유의한 상관관계가 존재하였다. 즉, $p21^{WAF1/CIP1}$과 p34의 기본 발현수준이 낮은 경우에 방사선 감수성이 높고, 기본 발현수준이 높은 경우에는 방사선 감수성이 낮은 상관관계가 존재함을 알 수 있었다. 결론: 방사선에 의해 유도된 아포토시스의 정도로 종양의 방사선 감수성을 예측하여 볼 수 있을 것으로 생각하며, 종양의 방사선 감수성을 예측할 수 있는 생물학적 표지자로 $p21^{WAF1/CIP1}$와 p34의 기본 발현수준이 이용될 수 있을 것으로 생각한다.

• 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.

• Molecular & Cellular Toxicology
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• 제3권1호
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• pp.31-35
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• 2007

Hesperidin, known as a flavonoid constituent of citrus, has been known to reduce the proliferation of several cancer cells. We investigated whether hesperidin-induced cell death on SNU-668, human gastric cancer cells. The cytotoxicity of hesperidin on SNU-668 cells was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay at the concentration of 1, 10, 50, and 100 ${\mu}M$. Cell viability by hesperidin was 53.18$\pm$2.85% of control value at 100 ${\mu}M$. The cell death by hesperidin showed apoptotic features, which were confirmed using a combination of 4, 6-diamidino-2-phenylindole (DAPI) staining and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay. In the apoptosis-regulating genes, treatment of hesperidin decreased mRNA expression of B-cell CLL/lymphoma 2 (BCL2), whereas expression of BCL2-associated X protein (BAX) was increased. The mRNA expression and the activity of caspase3 (CASP3), a major apoptotic factor, was significantly increased by hesperidin treatment. These results suggest that hesperidin could induce apoptosis through CASP3 activation on SNU-668, human gastric cancer cells.

• 한방안이비인후피부과학회지
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• 제22권1호
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• pp.11-32
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• 2009

Objective : This study was designed to investigate anti-cancer and whitening activities (LC). So it was investigated the effects of LC on proliferation rates of melanoma genetic profile by LC. Methods : The genetic profile for the effect of LC on human derived melanoma cell, SK-MEL-2, was measured using microarray technique, and the functional analysis on these genes were conducted. Total 441 genes were up-regulated and 830 genes down-regulated in cells treated with LC. Genes induced or suppressed by LC were all mainly concerned with basic signalling pathways, which are involved in cell growth, differentiation and migration. Especially, many genes, which are related in apoptosis and cell cycle arrest were up-regulated by treatment with LC, and genes related in cell cycle were down-regulated. Result : The network of total protein interactions were identified by using cytoscape program, and some key molecules, such as BCL2L1, SIN3A, SMAD2 and c-myc that can be used for elucidation of therapeutical mechanism of medicine in the future. Conclusion : These results suggest possibility of LC as addition drug and whitening cosmetics. In addition, it was also suggested that related mechanisms are involved in BCL2L1, SIN3A, SMAD2 and c-myc related signalling pathways.

• Journal of Chest Surgery
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• 제56권5호
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• pp.295-303
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• 2023

Background: The use of Adriamycin (ADR), also known as doxorubicin, as a chemotherapy agent is limited by its detrimental adverse effects, especially cardiotoxicity. Recent studies have emphasized the crucial role of angiotensin II (Ang-II) in the development of ADR-induced cardiomyopathy. This study aimed to explore the potential cardioprotective effects of losartan in a rat model of ADR-induced cardiomyopathy. Methods: Male Sprague-Dawley rats were randomly divided into 3 groups: a control group (group C), an ADR-treated group (ADR 5 mg/kg/wk for 3 weeks via intraperitoneal injections; group A), and co-treatment of ADR with losartan group (same dose of ADR and losartan; 10 mg/kg/day per oral for 3 weeks; group L). Western blot analysis was conducted to demonstrate changes in brain natriuretic peptide, collagen 1, tumor necrosis factor (TNF)-α, interleukin-6, matrix metalloproteinase (MMP)-2, B-cell leukemia/lymphoma (Bcl)-2, Bcl-2-associated X (Bax), and caspase-3 protein expression levels in left ventricular (LV) tissues from each group. Results: Losartan administration reduced LV hypertrophy, collagen content, and the expression of pro-inflammatory factors TNF-α and MMP-2 in LV tissue. In addition, losartan led to a decrease in the expression of the pro-apoptotic proteins Bax and caspase-3 and an increase in the expression of the anti-apoptotic protein Bcl-2. Moreover, losartan treatment induced a reduction in the apoptotic area compared to group A. Conclusion: In an ADR-induced cardiomyopathy rat model, co-administration of ADR with losartan presented cardioprotective effects by attenuating LV hypertrophy, pro-inflammatory factors, and apoptosis in LV tissue.

• 동의생리병리학회지
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• 제17권1호
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• pp.85-90
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• 2003

We investigated the effects of Oak smoke flavoring (OSF, Holyessing) on the growth of DU145 and PC-3 human prostate carcinoma cells. OSF treatment resulted in a concentration-dependent growth inhibition in both DU145 and PC3 cell lines. The anti-proliferative effect of OSF treatment was associated with the induction of apoptotic cell death which was confirmed by morphological change such as membrane shrinking, rounding up and chromatin condensation in DU145 and PC-3 cells. DNA flow cytometry analysis confirmed that OSF treatment increased population of apoptotic sub-G1 phase. Furthermore, we observed an increase of pro-apoptotic protein Bax expression and a decrease of anti-apoptotic protein Bcl-2 by OSF treatment in a dose-dependent manner. OSF also induced a proteolytic cleavage of specific target proteins such as poly(ADP-ribose) polymerase (PARP) and β-catenin proteins. The present results indicated that OSF-induced inhibition of human prostate carcinoma cell proliferation is associated with the induction of apoptosis.

• Archives of Pharmacal Research
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• 제24권4호
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• pp.342-348
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• 2001

In a course of obtaining more amount of bioactive costunolide and successive phytochemical isolation from Magnolia sieboldii (Magnoliaceae), a novel acyclic monoterpene 1 named deoxygeraniol (2,6(E)-dimethyl-2,6-octadiene) was isolated along with $\beta$-sitosterol 3-O-linoleate (2), trilinolein (3) and high amount of costunolide (4) in the pure state. The structure of compound 1 was determined on the basis of spectroscopic data. Costunolide was found to induce apoptotic cell death in a dose-dependent manner by nucleosomal DNA ladder and flow cytometric analysis. Immunoblot analysis showed that the level of the anti-apoptotic protein, Bcl-2, was decreased, whereas the cleavage of poly-(ADP-ribose) polymerase was, activated furthermore, the N-acetyl-L-cysteine antioxidant effectively prevented costunolide-induced cytotoxicity. These results suggest that costunolide-induced cell death is mediated by reactive oxygen species.