• Tuberculosis and Respiratory Diseases
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• v.65 no.6
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• pp.476-486
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• 2008

Background: TRAIL (TNF-related apoptosis inducing ligand) is a newly identified member of the TNF gene family which appears to have tumor-selective cytotoxicity due to the distinct decoy receptor system. TRAIL has direct access to caspase machinery and induces apoptosis regardless of p53 phenotype. Therefore, TRAIL has a therapeutic potential in lung cancer which frequently harbors p53 mutation in more than 50% of cases. However, it was shown that TRAIL also could activates $NF-{\kappa}B$ in some cell lines which might inhibit TRAIL-induced apoptosis. This study was designed to investigate whether TRAIL can activate $NF-{\kappa}B$ in lung cancer cell lines relatively resistant to TRAIL-induced apoptosis and inhibition of $NF-{\kappa}B$ activation using proteasome inhibitor MG132 which blocks $I{\kappa}B{\alpha}$ degradation can sensitize lung cancer cells to TRAIL-induced apoptosis. Methods: A549 (wt p53) and NCI-H1299 (null p53) lung cancer cells were used and cell viability test was done by MTT assay. Apoptosis was confirmed with Annexin V assay followed by FACS analysis. To study $NF-{\kappa}B$-dependent transcriptional activation, a luciferase reporter gene assay was used after making A549 and NCI-H1299 cells stably transfected with IgG ${\kappa}-NF-{\kappa}B$ luciferase construct. To investigate DNA binding of $NF-{\kappa}B$ activated by TRAIL, electromobility shift assay was used and supershift assay was done using anti-p65 antibody. Western blot was done for the study of $I{\kappa}B{\alpha}$ degradation. Results: A549 and NCI-H1299 cells were relatively resistant to TRAIL-induced apoptosis showing only 20~30% cell death even at the concentration 100 ng/ml, but MG132 ($3{\mu}M$) pre-treatment 1 hour prior to TRAIL addition greatly increased cell death more than 80%. Luciferase assay showed TRAIL-induced $NF-{\kappa}B$ transcriptional activity in both cell lines. Electromobility shift assay demonstrated DNA binding complex of $NF-{\kappa}B$ activated by TRAIL and supershift with p65 antibody. $I{\kappa}B{\alpha}$ degradation was proven by western blot. MG132 completely blocked both TRAIL-induced $NF-{\kappa}B$ dependent luciferase activity and DNA binding of $NF-{\kappa}B$. Conclusion: This results suggest that inhibition of $NF-{\kappa}B$ can be a potentially useful strategy to enhance TRAIL-induced tumor cell killing in lung cancer.

• Journal of Life Science
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• v.24 no.6
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• pp.595-602
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• 2014

Styela Clava tunic (SCT) has found some applications in many areas of medical treatment including as an anti-inflammatory compound, a wound healing film, in guided bone regeneration, and as a food additive. The protective effect of SCT aqueous extract (AE-SCT) on cell death induced by $H_2O_2$ treatment was investigated by measuring the changes in cell viability in HepG2 cells after AE-SCT treatment. High concentrations of antioxidant compounds including flavonoids (3.3 mg/g) and phenolics (32.3 mg/g) were detected in AE-SCT but no significant cytotoxicity was observed in HepG2 cells treated with AE-SCT. The viability of HepG2 cells was also not changed by treatment with different concentrations of AE-SCT after $H_2O_2$ treatment. However, cell viability was significantly increased in cells treated with three different concentrations of AE-SCT before $H_2O_2$ treatment. The greatest increase in cell viability was observed in the group treated with $50{\mu}g/ml$ AE-SCT, when compared with vehicle-treated group. FACS and DAPI staining analysis indicated that the decrease in number of dead cells was dependent on the concentration of AE-SCT. Alterations in the Bax/Bcl-2 ratio after $H_2O_2$ treatment were significantly restored by treatment with different concentrations of AE-SCT. These results indicate that AE-SCT, which contains high levels of antioxidants, may protect cells against death induced by $H_2O_2$ treatment.

• Journal of Life Science
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• v.22 no.4
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• pp.492-498
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• 2012

To investigate whether phytochemicals affect cancer cell viability, human colorectal HCT116 cells were treated with four different phytochemicals. Among these phytochemicals, curcumin is the strongest inhibitor of cell proliferation. In addition, it decreased cell viability in a dose-dependent manner. To unveil the molecular mechanisms involved in the inhibition of cell proliferation by curcumin, we carried out oligo DNA microarray analysis. We found that 137 genes were up-regulated more than 2-fold, and 141 genes were down-regulated more than 2-fold by 25 ${\mu}M$ curcumin treatment. Among the up-regulated genes, we selected 3 genes (ATF-3, GADD45A, and NR4A1) to confirm microarray data. The results of RT-PCR strongly agreed with those of the microarray data. Among the phytochemicals used in this study, curcumin is the strongest inducer of ATF3 expression, and increased ATF3 expression in a dose-dependent manner. Interestingly, FACS analysis showed that the inhibition of cell growth by curcumin was recovered by ATF3-siRNA transfection. Finally, we detected the changes of gene expression by ectopic expression of ATF3. The results indicated that many up-regulated genes were related to apoptosis. Overall, these results suggest that ATF3 may play an important role in the anti-proliferative activity of curcumin in human colorectal cancer cells.

• Journal of Korean Neurosurgical Society
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• v.48 no.1
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• pp.20-30
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• 2010

Objective : We determined whether the expression of GRIM-19 is correlated with pathologic types and malignant grades in gliomas, and determined the function of GRIM-19 in human gliomas. Methods : Tumor tissues were isolated and frozen at $-80^{\circ}C$ just after surgery. The tissues consisted of normal brain tissue (4), astrocytomas (2), anaplastic astrocytomas (2), oligodendrogliomas (13), anaplastic oligodendrogliomas (11), and glioblastomas (16). To profile tumor-related genes, we applied RNA differential display using a $Genefishing^{TM}$ DEG kit, and validated the tumor-related genes by reverse transcription polymerase chain reaction (RT-PCR). A human glioblastoma cell line (U343MG-A) was used for the GRIM-19 functional studies. The morphologic and cytoskeletal changes were examined via light and confocal microscopy. The migratory and invasive abilities were investigated by the simple scratch technique and Matrigel assay. The antiproliferative activity was determined by thiazolyl blue Tetrazolium bromide (MTT) assay and FACS analysis. Results : Based on RT-PCR analysis, the expression of GRIM-19 was higher in astrocytic tumors than oligodendroglial tumors. The expression of GRIM-19 was higher in high-grade tumors than low-grade tumors or normal brain tissue; glioblastomas showed the highest expression. After transfection of GRIM-19 into U343MG-A, the morphology of the sense-transfection cells became larger and more spindly. The antisensetransfection cells became smaller and rounder compared with wild type U343MG-A. The MTT assay showed that the sense-transfection cells were more sensitive to the combination of interferon-$\beta$ and retinoic acid than U343MG-A cells or antisense-transfection cells; the antiproliferative activity was related to apoptosis. Conclusion : GRIM-19 may be one of the gene profiles which regulate cell death via apoptosis in human gliomas.

• The Journal of Korean Obstetrics and Gynecology
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• v.24 no.3
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• pp.1-13
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• 2011

Objectives: This study was designed to investigate the analysis of apoptosis by Scirpi Tuber in Hela cell and MCF-7 cell. Methods: For cytotoxic effect of Scirpi Tuber extract, Scirpi Tuber extract were cultured on NIH3T3 cell in vitro. After treatment with various concentration of Scirpi Tuber, cell growth was evaluated in Hela cell and MCF-7 cell. Hoechst 33342 staining was performed to estimate DNA fragment effect of Scirpi Tuber on the apoptosis in Hela cell and MCF-7 cell. Annexin V/PI apoptosis assay was used to estimate the effects of Scirpi Tuber on the early apoptosis in Hela cell MCF-7 cell. All the stained cells were analyzed by a FACS. RT-PCR was used to estimate the apoptosis gene expression effect of Scirpi Tuber extract on Hela cell and MCF-7 cell. Results: Cytotoxic effect of Scirpi Tuber extract was not found on per NIH3T3 cell. The viability of Hela cell was significantly decreased Scirpi Tuber (500, $1000{\mu}g/m\ell$) in Hela cell 1day, 3day and 5days after treatment (p<0.01). The viability of MCF-7 cell was significantly decresed Scirpi Tuber ($1000{\mu}g/m\ell$) in MCF-7 cell (p<0.01), Scirpi Tuber ($500{\mu}g/m\ell$) in MCF-7 cell only 3days after treatment (p<0.01). In RT-PCR analysis, after treatment of $100{\mu}g/m\ell$ of ACR extract, BCL-2 were decreased and BAX, caspase-3 were increased both in Hela cell and MCF-7 cell. DNA fragmentation was observed the Scirpi Tuber on Hela cell and MCF-7 cell. As time goes on DNA fragmentation incresed. In Annexin V/PI apoptosis assay, after treatment of $1mg/m\ell$ of Scirpi Tuber, the early apoptotic cell increased both in Hela cell and MCF-7 cell. As time goes on apoptotic cell increased. Conclusion: Scirpi Tuber appears to have considerable activity on the apoptosis in Hela cell and MCF-7 cell.

• Science of Emotion and Sensibility
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• v.16 no.1
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• pp.95-106
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• 2013

The study compares accuracies between humans and computer algorithms in the discrimination of spontaneous smiles from posed smiles. For this purpose, subjects performed two tasks, one was judgment with single pictures and the other was judgment with pair comparison. At the task of judgment with single pictures, in which pictures of smiling facial expression were presented one by one, subjects were required to judge whether smiles in the pictures were spontaneous or posed. In the task for judgment with pair comparison, in which two kinds of smiles from one person were presented simultaneously, subjects were to select spontaneous smile. To calculate the discrimination algorithm accuracy, 8 kinds of facial features were used. To calculate the discriminant function, stepwise linear discriminant analysis (SLDA) was performed by using approximately 50 % of pictures, and the rest of pictures were classified by using the calculated discriminant function. In the task of single pictures, the accuracy rate of SLDA was higher than that of humans. In the analysis of accuracy on pair comparison, the accuracy rate of SLDA was also higher than that of humans. Among the 20 subjects, none of them showed the above accuracy rate of SLDA. The facial feature contributed to SLDA effectively was angle of inner eye corner, which was the degree of the openness of the eyes. According to Ekman's FACS system, this feature corresponds to AU 6. The reason why the humans had low accuracy while classifying two kinds of smiles, it appears that they didn't use the information coming from the eyes enough.

• Journal of Life Science
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• v.21 no.6
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• pp.851-857
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• 2011

Sulforaphane (SFN) is an isothiocyanate, isolated from glucoraphanin in broccoli and other cruciferaous vegetables. Recent studies have revealed that SFN induces anti-proliferation and apoptosis by cell cycle arrest in various cancer cells. In this study, we investigated the effect of SFN induced apoptosis in chondrosarcoma HTB-94 cells. SFN caused suppression of proliferation and apoptosis in a dose-dependent manner as determined by cell phenotype, MTT assay and FACS analysis in HTB-94 cells. Treatment of SFN led to caspase-3 activation and p53 accumulation as determined by Western blot analysis. Also, SFN significantly induced DNA fragmentation and nuclear degradation though activation of caspase-3, as detected by DNA electrophoresis and immunostaining, respectively. Our results indicate that SFN-induced apoptosis was regulated by p53 and caspase-3 dependent pathways. Furthermore, SFN may act as a potent anti-proliferation agent, and as a promising candidate for molecular-targeting chemotherapy against human chondrosarcoma cells.

• Journal of the Korean Applied Science and Technology
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• v.38 no.5
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• pp.1302-1313
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• 2021

In this study, Gami-Sumiwon (GS; Cynanchi wilfordii R., Angelica gigantis R., Lycium chinense M., Betula platyphylla S., Cryptotympana atrata F. and Carthamus tinctorius L.) and/or Sinomenium acutum R. (SC) was extracted with 70% ethanol or water. And We investigated the antioxidant activity effect of GS±SC. The following experimental techniques were used to evaluate the antioxidant efficacy of GS±SC. HPLC chromatogram, heavy metal content, ABTS/DPPH radical scavenging analysis, SOD-like activity assay, FACS, and NO assay. As a result of the experiment, the sinomenine content was found to be higher in DW extracts, and decursin was found to be higher in 80% ethanol extracts. And, the amount of heavy metals in all extracts was below the standard value. ABTS, and DPPH radical scavenging activity was identified that GS±SC(EtOH) was found to have a higher scavenging activity than GS±SC(DW). But, SOD showed the opposite result. No cytotoxicity of GS was observed on Raw 264.7 cells at concentration of 1~100 ㎍/㎖. The ROS production was significantly decreased that GS±SC(DW) was found to have a higher scavenging activity than GS±SC(EtOH). However, NO production showed the opposite result. Looking at the results of SOD and ROS analysis, SC does not seem to have a function of prevention. SC is thought to have an effect on the removal of free radicals generated after oxidative stress. This result objectively confirmed the antioxidant effect of GS±SCs. We will continue to conduct in-depth research. Therefore, it is believed that the possibility of using GS±SCs as a functional material can be established. The more diverse the objectives, the higher the value of GS utilization is thought to be.

• Journal of Acupuncture Research
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• v.17 no.1
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• pp.221-250
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• 2000

In order to investigate experimentally that the effect of aqua-acupuncture solution of Paeonia lactiflora on arthritis of mice induced by collagenII, the author performed several experimental items : that is increase, paw thickness, DTH, weight of spleen, hematological change, expression of $CD4^+$, $CD8^+$, $CD19^+$, gene expression of IL-1, IL-6, IL-12, IFN-${\gamma}$, TNF, proliferation of synovial cells and cytotoxicity. The results were obtained as follows; 1. Inhibitory effects of aqua-acupuncture solution of Paeonia lactiflora on arthritis induced by collgenII. 1) In incidence, paw edema, AI and DTH were inhibited as compared with control group. 2) The splenic weight was increased and the number of leukocytes was decreased as compared with control group. 3) The number of $CD4^+$, $CD8^+$ activated cells and surface-receptor expression were increased as compared with control group. 4) In hematological change, total protein, creatinine and LDH were decreased significantiy as compared with control group. 2. FACS analysis on normal BABL/c of spenic cells treated with aqua-acupuncture solution of Paeonia lactiflora. 1) Aqua-acupuncture solution of Paeonia lactiflora activated adhesive splenic cells of mice morphologically in vitro. 2) Aqua-acupuncture solution of Paeonia dactiflora enhanced the gene expression of IL-12 and also enhanced that of interferon-${\gamma}$ remarkably. 3) Aqua-acupuncture solution of Paeonia lactiflora reduced the number of $CD4^+$, $CD8^+$, $CD19^+$ activated cells and their surface-receptor expression as compared with control group. 3. Effects of aqua-acupuncture solution of Paeonia lactiflora on human synovial cells. 1) In cytotoxicity against synovial cells, aqua-acupuncture solution of Paeonia lactiflora didn't show cytotoxicity at concentration of $10-100{\mu}g/m{\ell}$ but showed significantly at concentration of $200-400{\mu}g/m{\ell}$ as compared with control group. 2) Aqua-acupuncture solution of Paeonia lactiflora reduced the gene expression of IL-6, IL-$1{\beta}$ and TNF-${\alpha}$. 3) Aqua-acupuncture solution of Paeonia lactiflora inhibited proliferation of synovial cells at concentration of 100 and $200{\mu}g/m{\ell}$.

• The Journal of Dong Guk Oriental Medicine
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• v.9
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• pp.51-71
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• 2000

The present experiments were designed to study the influence of WalgookwhanhabBojoongikgitang on immune functions of Balb/c mice under stress condition. WalgookwhanhabBojoongikgitang was orally administered to the mice for 15 days. On the 10th day the mice were immunized with sheep red blood cells (SRBC) and then subjected to electric footshock for 5 days (2 sessions a day, 11 footshocks a 31 min-session). The immune responses to SRBC were determined by means of hemagglutination and BIT cell populations in the spleen were studied by FACS analysis on the 16th day. The results were as follows. 1. After electric footshock, IDlce became sluggish and crowded to one side of the cage. Increased antibody titer for SRBC, increased B cell population, and decreased T cell population in the spleen were also observed. These results confirm that electric footshock caused stress inducing immunological and behavioral changes in Balblc mice. 2. Walgookwhanhab-Bojoongikgitang administration significantly antagonized the effect of electric footshock on the antibody titer for SRBC. As a result, antibody titers for SRBC in the mice treated with Walgookwhanga-Bojoongikgitang were maintained at the similar levels as those of control group mice even after the electric foots hock. 3. Walgookwhanhab-Bojoongikgitang administration also antagonized the effect of electric footshock on the BIT populations in spleen. As a result, Band T populations in the mice treated with Walgookwhanhab-Bojoongikgitang were maintained at the similar levels as in the control group mice even after the electric footshock. Taken together, Walgookwhanhab-Bojoongikgitang seem to help Balb/c mIce to maintain their humoral immune response and immune cell populations at a normal range under the stress conditions, suggesting its possible therapeutic use as a immune function modulator.