• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.983-987
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• 2014

Nigella sativa (N sativa), commonly known as black seed, has been used in traditional medicine to treat many diseases. The antioxidant, anti-inflammatory, and antibacterial activities of N sativa extracts are well known. Therefore, the present study was designed to investigate the anticancer activity of seed extract (NSE) and seed oil (NSO) of N sativa against a human lung cancer cell line. Cells were exposed to 0.01 to 1 mg/ml of NSE and NSO for 24 h, then percent cell viability was assessed by 3-(4, 5-dimethylthiazol-2yl)-2, 5-biphenyl tetrazolium bromide (MTT) and neutral red uptake (NRU) assays, and cellular morphology by phase contrast inverted microscopy. The results showed NSE and NSO significantly reduce the cell viability and alter the cellular morphology of A-549 cells in a concentration dependent manner. The percent cell viability was recorded as 75%, 50%, and 26% at 0.25, 0.5, and 1 mg/ml of NSE by MTT assay and 73%, 48%, and 23% at 0.25, 0.5, and 1 mg/ml of NSE by NRU assay. Exposure to NSO concentrations of 0.1 mg/ml and above for 24 h was also found to be cytotoxic. The decrease in cell viability at 0.1, 0.25, 0.5, and 1 mg/ml of NSO was recorded to be 89%, 52%, 41%, and 13% by MTT assay and 85%, 52%, 38%, and 11% by NRU assay, respectively. A-549 cells exposed to 0.25, 0.5 and 1 mg/ml of NSE and NSO lost their typical morphology and appeared smaller in size. The data revealed that the treatment of seed extract (NSE) and seed oil (NSO) of Nigella sativa significantly reduce viability of human lung cancer cells.

• Toxicological Research
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• 제19권1호
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• pp.45-50
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• 2003

The purpose of this study was to determine the role of substituted groups in phenolic compounds to develop an anticancer agent having strong cytotoxicity against cancer cells but weak against normal cells. The phenolic compounds used in this study were gallic acid and ferulic acid with hydroxyl and carboxyl groups, syringic acid with hydroxyl, carboxyl and methoxy groups, and pyre-gallol with hydroxyl groups. Cytotoxicities of these compounds were evaluated by MTT assay for cell viability and XTT assay for cell adhesion activity in normal human skin fibroblast (Detroit 551) and human skin melanoma (SK-MEL-3) cells. Syringic acid, gallic acid and ferulic acid decreased the cell viability and cell adhesion activity in SK-MEL-3 cells but not in Detroit 551 cells while pyrogallol decreased in both cells. The susceptibility of cell viability based on the $IC_{50}$ values of MTT assay in Detroit 551 cells was in the following order: pyrogallol > gallic acid > ferulic acid > syringic acid, while it was in SK-MEL-3 cells: Syringic acid > progallol > ferulic acid > gallic acid. These results suggest that carboxyl and methoxy groups of these compounds play an important role in selectivity of cytotoxicity in normal and cancer cells.

• 생명과학회지
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• 제18권9호
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• pp.1312-1315
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• 2008

DAPI 염색을 이용하여 세포사멸 유도물질의 초고속스크리닝에 적합한 protocol을 제작하였다. Liquid handler에서 dispensing과 multilabel counter에서의 상대적인 fluorescent intensity를 측정하는 과정을 자동화한 protocol을 제작하였고, 이의 과정은 매우 효율적으로 가동되었음을 확인하였다. DAPI에 binding된 DNA의 intensity를 측정하는 조건검토에서 가장자리의 36개를 reading하지 않는 것이 edge effect를 줄일수 있는 것을 확인하였고, 0.1 sec reading으로 시간을 절약할 수 있었다. 이의 결과를 효과적으로 이용할 경우, 세포사멸 유도물질의 초고속스크리닝 또는 이에 적합한 스크리닝이 가능함을 확인하였다.

• 대한한의학회지
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• 제24권2호
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• pp.159-165
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• 2003

Objectives : Coptidis rhizoma (CR) is an oriental medicine that has been used in many traditional prescriptions against diabetes mellitus in Korea for centuries. Our purpose was to determine the protective effect and its action mechanism of CR on the cytotoxicity of pancreatic -cell line (RINm5F cell). Methods : In this experiment, we used methods such as MTT assay for detection of cytotoxicity, DNA fragmentation assay for detection of apoptotic cell death, LDH activity assay for detection of necrotic cell death, and measurement of $DiOC_{6}$ (3) retention for detection of mitochondrial membrane potential (MMP). Background : Nitric oxide (NO) is believed to playa key role in the process of pancreatic -cell destruction leading to insulin-dependent diabetes mellitus (IDDM). Results : Exposure of RINm5F cells to chemical NO donor such as S-nitroso-N-acetylpenicillamine (SNAP) induced cytotoxic events such as DNA fragmentation and lactate dehydrogenase (LDH) release into medium. However, pretreatment of RINm5F cells with CR extract ($10~50{\mu\textrm{g}}/ml$) for 3 hours prevented SNAP-induced DNA fragmentation and LDH release into medium through the inhibition of MMP disruption. Conclusions : These results suggest that CR may be a candidate for a therapeutic or preventing agent against IDDM.

• 대한한방종양학회지
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• 제7권1호
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• pp.1-18
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• 2001

Objective: This experimental study was carried out to evaluate the effects of Saussurea Radix and Plantaginis Herba on cellular viability, proliferation, apoptosis and expression of the cell cycle-related genes in cultured gastric cancer cells. Method :MTT assay for analysis of cellular toxicity and the effect on suppression of cellular viability, $[^{3}H]$ thymidine incorporation assay for evaluation of the effect on suppression of DNA replication, tryphan blue exclusion assay for measurement of induction of apoptosis and Quantitative RT-PCR for analysis of the effects on expression of cell cycle or apoptosis-related genes were performed. Results: Antitumor activity of Saussurea Radix associated with inhibition of cell cycle progression and promotion of apoptosis caused by transcriptional regulation of p53, p21/Wafl and the other related genes was observed.

• Biomolecules & Therapeutics
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• 제19권1호
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• pp.134-139
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• 2011

Sphingosine kinase (SPHK) has a central role to control cell death and cell proliferation, which is suggested as a sphingolipid rheostat by regulating the levels between ceramide and sphingosine 1-phosphate (S1P). Therefore, physiological regulators of SPHK will be a good candidate to develop a new targeted drug. For this purpose, a series of synthetic pseudoceramides were tested by SPHK assay either cell-based or cell-free system. K10PC-5 strongly inhibited SPHK, while K6PC-5 activated SPHK in cell-free system. Specifically, K6PC-5 activated SPHK under the co-treatment with $50\;{\mu}M$ dimethylsphingosine (DMS), a SPHK inhibitor. Collectively, we developed a simple SPHK assay system to find SPHK regulatory pseudoceramide compounds, K10PC-5 and K6PC-5 which may be useful to cancer treatment or immune regulation like FTY720, a synthetic sphingolipid mimetic compound.

• Tuberculosis and Respiratory Diseases
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• 제55권5호
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• pp.488-498
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• 2003

연구배경 : 세포에 방사선을 조사하면 세포사멸과 함께 AP-l, NF-${\kappa}B$와 같은 여러 전사인자가 활성화 되는 것으로 알려져 있다. 이중 염증과 면역반응의 중요한 전사인자인 NF-${\kappa}B$는 항아포프토시스의 기능이 있으며 NF-${\kappa}B$ 활성화를 차단함으로써 TNF-${\alpha}$나 daunorubicine 등에 의 한 항암효과를 상승시킬 수 있음이 밝혀져 있다. NF-${\kappa}B$의 항아포프토시스 기전은 NF-${\kappa}B$ 의존성 단백질인 cIAPI, 2의 전사발현 유도에 의한 것으로 cIAPl, 2는 caspase 3, 7, pro-caspase-9의 활성을 차단함으로써 아포프토시스를 억제하는 것으로 알려져 있다. 이에 저자들은 방사선 유도성 세포사멸에 비교적 내성을 보이는 A549 세포주에서 방사선에 의한 NF-${\kappa}B$ 활성화와 그에 따른 cIAP 발현유도를 조사하고 NF-${\kappa}B$ 활성화를 차단하여 방사선 유도성 세포사멸의 감작효과를 확인하기 위하여 본 연구를 시행하였다. 방 법 : 세포주는 A549 폐암세포주를 이용하였고, 방사선 조사는 Varian사의 Clinac 1800C 선형가속기를 이용하였으며 조사량은 10GY를 사용하였다. 세포독성 검사는 MTT Assay를 이용하였고, NF-${\kappa}B$ 활성화 검사는 luciferase reporter gene assay, electromobility shift assay, $I{\kappa}B-{\alpha}$ degradation에 대한 westem blot을 이용하였다. NF-${\kappa}B$활성을 차단하기 위하여 proteosome inhibitor인 MGI32와 $I{\kappa}B{\alpha}$-superrepressor plasmid를 transfection한 안정적 세포주 A549-$I{\kappa}B{\alpha}$-superrepressor를 이용하였다. cIAP의 발현은 RT-PCR을 이용하였고, cIAP2 promoter 활성은 NF-${\kappa}B$ site를 포함한 cIAP2 유전자 5' f1anking region(1.4kb)을 pGL2-Basic luciferase vector에 cloning 한 construct를 사용하여 transfection 후 luciferase assay를 시행하였다. 결 과 : A549 cell에서 10Gy 방사선 조사에 의한 세포독성은 24hr, 48hr에 각각 $10.82{\pm}.3%$, $17.7{\pm}6.4%$로 비교적 내성이 있음을 확인하였다. 방사선에 의한 NF-${\kappa}B$의 활성은 $I{\kappa}B-{\alpha}$ 분해에 대한 western blot과 EMSA로 확인하였으며 luciferase assay에서도 약 1.6 배 정도의 NF-${\kappa}B$ 활성화가 있었다. NF-${\kappa}B$활성을 차단하기 위해 사용한 MG132는 방사선 유도성 세포사멸에 영향을 주지 않았으며 A549-$I{\kappa}B{\alpha}$-superrepressor 세포주에서도 세포사멸의 감작효과는 없었다. 또한 RT-PCR 결과 방사선에 의한 cIAP1,2 mRNA 발현유도는 관찰되지 않았고 cIAP2 promoter luciferase assay에서도 cIAP2 전사활성 유도는 없었다. 결 론 : A549 폐암세포주에서 방사선에 의해 NF-${\kappa}B$의 활성화는 확인하였으나 활성화 정도가 미약하였고 NF-${\kappa}B$ 의존성 항아포프토시스 유전자인 cIAP가 방사선에 의해 발현유도 되지 않았으며, NF-${\kappa}B$ 활성을 차단함에도 세포독성에 감작효과가 없었으므로 A549 폐암세포주에서 방사선 유도성 세포사멸에 내성을 보이는 기전에는 NF-${\kappa}B$의 역할이 미미하리라고 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권1호
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• pp.131-133
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• 2016

Background: Combination chemotherapy regimes are common treatments for cancer. The aim of this study was to evaluation the effect of individual chemotherapeutic agents in comparison with a first line chemotherapy regime treatment in the AGS gastric cancer cell line by MTT assay. Materials and Methods: In this experimental study, AGS cells were grown in RPMI-1640 supplemented with 10% fetal calf serum and 100 IU/ml penicillin, and $10{\mu}g/ml$ streptomycinin, under a humidified condition at $37^{\circ}C$ with 5% CO2. All cells were washed with PBS and detached with trypsin, centrifuged and 8000 cells re-plated on to 96- well plates. LD50 doses of Epirubicin, Cisplatin and 5-fluorouracil were added to each well in mono or triple therapy. Anti-proliferative activities were determined by MTT assay after 24, 48 or 72 h. Results: Results of MTT assays showed that there were no significant differences among 3 drugs in monotherapy (p=0.088), but there was significant difference between combination therapy with epirubicin (P=0.031) and 5FU (p=0.013) on cell survival at 24 h. After 48 and 72 hours, cell viability showed significant differences between the 3 drugs (p=0.048 and P=0.000 for 48 and 72 h, respectively) and there was significant difference between combination therapy with epirubicin (P=0.035 and P=0.002 for 48 and 72 h, respectively). Conclusions: The results showed no significant differences between these chemotherapy drugs each given alone, but combination therapy with 3 drugs had significant effects on cell viability in comparison with epirubicin alone.

• 대한한방부인과학회지
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• 제22권1호
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• pp.125-139
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• 2009

Purpose: This experiment was designed to find out increasing effects of S. barbata. co-treatment with anti-cancer drugs at cancer cell's growth inhibition effect. Methods: Divergent observational study of the S. barbata. co-treatment with Cisplatin treatment on HeLa cell. Cell viability using MTT assay, Cell Culture and Cytotoxicity Studies, Cell Cycle Analysis, Annexin V-FITC/PI assay, Cell morphological assessment, PARP cleavage using Western blotting analysis when HeLa cell were co-treated with Cisplatin and Scutellaria Barbata extracts. Results: When HeLa cell were co-treated with Cisplatin and Scutellaria Barbata extracts, we found out viability of HeLa cell, changing in the distribution of cell cycle, Annexin V-FITC staining, DAPI staining, PARP clavage protein assay by Western-blot. So Scutellaria Barbata extracts have increased apoptosis Conclusion: When co-treated Scutellaria Barbata extracts with anti-cancer drugs, the anti-cancer effects were increased. We still not sure which constituent apoptosis at cancer cells and activates anti-cancer effects suppressing, but we believe that it'll be revealed here after with following experiments.

• 약학회지
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• 제52권5호
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• pp.412-417
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• 2008

BAY11-7082 was initially found to be an anti-inflammatory drug with NF-${\kappa}B$ inhibitory property. In this study, we evaluated modulatory function of BAY11-7082 on U937 cell-cell adhesion induced by CD29 (${\beta}1$-integrins). BAY11-7082 strongly blocked functional activation of CD29 (${\beta}1$-integrins), as assessed by cell-cell adhesion assay. However, this compound did not block a simple activation of CD29, as assessed by cell-fibronectin adhesion assay. In particular, to understand molecular mechanism of BAY11-7082-mediated inhibition, the regulatory roles of CD29-induced actin cytoskeleton rearrangement under cell-cell adhesion and surface level of CD29 were examined using confocal and flow cytometic analysis. Interestingly, this compound strongly suppressed the molecular association of actin cytoskeleton with CD29 at cell-cell adhesion site. Moreover, BAY11-7082 also diminished surface levels of CD29 as well as its-associated adhesion molecule CD147, but not other adhesion molecules such as CD18 and CD43. Therefore, our data suggest that BAY11-7082 may be involved in regulating immune responses managed by CD29-mediated cell-cell adhesion.