• Journal of Ginseng Research
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• v.43 no.4
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• pp.527-538
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• 2019

Background: Ginsenoside Rg1 was shown to exert ligand-independent activation of estrogen receptor (ER) via mitogen-activated protein kinase-mediated pathway. Our study aimed to delineate the mechanisms by which Rg1 activates the rapid ER signaling pathways. Methods: ER-positive human breast cancer MCF-7 cells and ER-negative human embryonic kidney HEK293 cells were treated with Rg1 ($10^{-12}M$, $10^{-8}M$), $17{\beta}$-estradiol ($10^{-8}M$), or vehicle. Immunoprecipitation was conducted to investigate the interactions between signaling protein and ER in MCF-7 cells. To determine the roles of these signaling proteins in the actions of Rg1, small interfering RNA or their inhibitors were applied. Results: Rg1 rapidly induced $ER{\alpha}$ translocation to plasma membrane via caveolin-1 and the formation of signaling complex involving linker protein (Shc), insulin-like growth factor-I receptor, modulator of nongenomic activity of ER (MNAR), $ER{\alpha}$, and cellular nonreceptor tyrosine kinase (c-Src) in MCF-7 cells. The induction of extracellular signal-regulated protein kinase and mitogen-activated protein kinase kinase (MEK) phosphorylation in MCF-7 cells by Rg1 was suppressed by cotreatment with small interfering RNA against these signaling proteins. The stimulatory effects of Rg1 on MEK phosphorylation in these cells were suppressed by both PP2 (Src kinase inhibitor) and AG1478 [epidermal growth factor receptor (EGFR) inhibitor]. In addition, Rg1-induced estrogenic activities, EGFR and MEK phosphorylation in MCF-7 cells were abolished by cotreatment with G15 (G protein-coupled estrogen receptor-1 antagonist). The increase in intracellular cyclic AMP accumulation, but not Ca mobilization, in MCF-7 cells by Rg1 could be abolished by G15. Conclusion: Ginsenoside Rg1 exerted estrogenic actions by rapidly inducing the formation of ER containing signalosome in MCF-7 cells. Additionally, Rg1 could activate EGFR and c-Src ER-independently and exert estrogenic effects via rapid activation of membrane-associated ER and G protein-coupled estrogen receptor.

• Tuberculosis and Respiratory Diseases
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• v.50 no.1
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• pp.52-66
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• 2001

Background : NF-${\kappa}B$ is the most important transcriptional factor in IL-8 gene expression. Triptolide is a new compound that recently has been shown to inhibit NF-${\kappa}B$ activation. The purpose of this study is to investigate how triptolide inhibits NF-${\kappa}B$-dependent IL-8 gene transcription in lung epithelial cells and to pilot the potential for the clinical application of triptolide in inflammatory lung diseases. Methods : A549 cells were used and triptolide was provided from Pharmagenesis Company (Palo Alto, CA). In order to examine NF-${\kappa}B$-dependent IL-8 transcriptional activity, we established stable A549 IL-8-NF-${\kappa}B$-luc. cells and performed luciferase assays. IL-8 gene expression was measured by RT-PCR and ELISA. A Western blot was done for the study of $I{\kappa}B{\alpha}$ degradation and an electromobility shift assay was done to analyze NF-${\kappa}B$ DNA binding. p65 specific transactivation was analyzed by a cotransfection study using a Gal4-p65 fusion protein expression system. To investigate the involvement of transcriptional coactivators, we perfomed a transfection study with CBP and SRC-1 expression vectors. Results : We observed that triptolide significantly suppresses NF-${\kappa}B$-dependent IL-8 transcriptional activity induced by IL-$1{\beta}$ and PMA. RT-PCR showed that triptolide represses both IL-$1{\beta}$ and PMA-induced IL-8 mRNA expression and ELISA confirmed this triptolide-mediated IL-8 suppression at the protein level. However, triptolide did not affect $I{\kappa}B{\alpha}$ degradation and NF-$_{\kappa}B$ DNA binding. In a p65-specific transactivation study, triptolide significantly suppressed Gal4-p65T Al and Gal4-p65T A2 activity suggesting that triptolide inhibits NF-${\kappa}B$ activation by inhibiting p65 transactivation. However, this triptolide-mediated inhibition of p65 transactivation was not rescued by the overexpression of CBP or SRC-1, thereby excluding the role of transcriptional coactivators. Conclusions : Triptolide is a new compound that inhibits NF-${\kappa}B$-dependent IL-8 transcriptional activation by inhibiting p65 transactivation, but not by an $I{\kappa}B{\alpha}$-dependent mechanism. This suggests that triptolide may have a therapeutic potential for inflammatory lung diseases.

• BMB Reports
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• v.30 no.5
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• pp.303-307
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• 1997

Grb2, which is composed of a Src homology 2 (SH2) domain and two Src homology 3 (SH3) domains, is known to serve as an adaptor protein in signaling for Ras activation. Thus, a blocker of the Grb2 interactions with other proteins can be a potential candidate for an anticancer drug. In this study, we have developed a high throughput screening method for SH3 domain binding ligands and blockers. Firstly, we made and purified the glutathione S-transferase (GST)-fusion proteins with the Grb2 SH2 and SH3 domains, and the entire Grb2. This method measures the binding of a biotin-labeled oligopeptide, derived from a Grb2/SH3 binding motif in the hSos, to the GST-fusion proteins, which are precoated as glutathione S-transferase fusion protein on a solid phase. When $1\;{\mu}g$ of each fusion protein was used to coat the wells, both N- and C- terminal SH3 the domains as well as the whole of Grb2 were able to interact with the biotin-conjugated ligand peptide, while the SH2 domain and GST alone showed no binding affinity. Although N- and C- terminal SH3 domains showed an increase of binding to the ligand peptide in proportion to the amount of peptide, the GST fusion protein with Grb2 demonstrated much higher binding affinity. GST-Grb2 coating on the solid phase showed a saturation curve; 66 and 84% of the maximal binding was observed at 100 and 300 ng/$100\;{\mu}l$, respectively. This binding assay system was peptide sequence-specific, showing a dose-dependent inhibition with the unlabeled peptide of SH3 binding motif. Several other peptides, such as SH2 domain binding motifs and PTB domain binding motif, were ineffective to inhibit the binding to the biotin-conjugated ligand peptide. These results suggest that our method may be useful to screen for new anticancer drug candidates which can block the signaling pathways mediated by SH3 domain binding.

• Korea Journal of Hospital Management
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• v.6 no.3
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• pp.148-166
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• 2001

The purpose of this study is to classify elderly patient in long-term care facilities using RUG(Resource Utilization Group)-III. It is designed by measuring patient medical characteristics and medical staff time. Elderly patients are classified into 7 categories by clinical(medical and behavioral) hierarchical typology of patients. Through the tertiary split, all 44 groups are formulated. This classification is explained by each patient resource(staff time) utilization level which is called CMI(Case-Mix Index). Major findings are as follows; 1. The objects in this study were classified into 35 groups out of 44 groups. The most frequent category is clinical complex category(CCC; 38.9%). And extensive service category(ESC; 18.8%), reduced physical function category(RPC; 13.1%), special rehabilitation category(SRC; 12.8%), and impaired cognitive category(ICC; 0.00%) are followed. 2. The mean of total CMI was $1.02{\pm}0.36$, ranging from 0.68 to 1.44(1 vs 2.12). The mean of CMI of SRC is only 1.17 which should be the highest. The means of ESC and see are equally 1.20. The means of CMI of CCI, ICC, BPC, and RPC were 0.90, 0.75, 0.83 and 0.96, respectively. 3. The validity of this classification was tested. Trend-test using Regression Analysis was done in the secondary split level. SCC, CCC, ICC, and RPC which covered 68.4% of this research objects showed linear trend of CMI in interim classification. This results were statistically significant. 4. In clinical hierarchy, the trend were showed linearity. But the multiple comparison of categories using Scheffe-test showed that SRC, ESC and see had same level of CMI means and CCC and ICC, too. This results were statistically significant. Classifying elderly patients with RUG-III, the results showed partly linear trend in clinical hierarchy and in interim classification in conclusion. But, in clinical hierarchy, it was failed to show the consistent order of CMI. It can be explained by two reasons. One is that this research subjects were overlapped in each clinical hierarchy group. And the other is that the some of the characteristics for clinical hierarchy is not appropriate for them. For the further study, it needs to have proper sample size and to modify RUG-III to K-RUG to consider our.. medical environment.

• International Journal of Oral Biology
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• v.39 no.3
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• pp.131-136
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• 2014

Porphyromonas gingivalis is one of the most important periodontal pathogens and has been to known to invade various types of cells, including endothelial cells. The present study investigated the mechanisms involved in the internalization of P. gingivalis in human umbilical vein endothelial cells (HUVEC). P. gingivalis internalization was reduced by clathrin and lipid raft inhibitors, as well as a siRNA knockdown of caveolin-1, a principal molecule of lipid raft-related caveolae. The internalization was also reduced by perturbation of actin rearrangement, while microtubule polymerization was not required. Furthermore, we found that Src kinases are critical for the internalization of P. gingivalis into HUVEC, while neither Rho family GTPases nor phosphatidylinositol 3-kinase are required. Taken together, this study indicated that P. gingivalis internalization into endothelial cells involves clathrin and lipid rafts and requires actin rearrangement associated with Src kinase activation.

• Proceedings of the Korean Information Science Society Conference
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• 2010.11a
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• pp.127-128
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• 2010

본 논문에서는 CCTV 감시 영상에서 취득한 얼굴 이미지를 이용하여, 범죄자 감시목록에 등록된 범죄 용의자를 탐지 식별하는 시스템을 설계 및 구현하였다. 특히 본 논문에서 제안한 SVDD와 SRC를 혼합한 계층적 구조의 범죄 용의자 식별 모듈은 다음과 같은 특성을 갖는다: 1) 먼저 SVDD를 이용하여 범죄 용의자만을 빠르게 인식함으로써, 일반인에 대한 불필요한 범죄자 식별 연산을 수행하지 않는다; 2) 다양한 식별 성능을 저해하는 환경에서도 이미 강인한 성능이 검증된 SRC를 범죄 용의자 식별과정에 적용함으로써 안정적이고 정확한 식별 시스템을 보장한다; 3) 동일 생체 특정의 반복적 사용을 통한 다수결 투표전략을 취함으로써 시스템의 신뢰도를 보장한다; 4) 점증적 갱신의 학습 능력으로 인하여 범죄 용의자 감시목록 데이터베이스의 변화에도 능동적으로 적응한다 실제 KUFD(Korea University Face Database)를 자체 제작하고 캠퍼스 내에서 CCTV 환경의 얼굴 인식 기반 범죄 용의자 탐지 및 식별 시스템 환경을 모의 구축하여 실험적으로 제안된 시스템의 성능을 검증한다.

• The Korean Journal of Physiology and Pharmacology
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• v.11 no.1
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• pp.31-36
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• 2007

To elucidate a potential molecular link between diabetes and atherosclerosis, we investigated the role of Janus tyrosine kinase(JAK) for NAD(P)H oxidase-derived superoxide generation in the enhanced proliferative capacity of vascular smooth muscle cells(VSMC) of Otsuka Long-Evans Tokushima Fatty(OLETF) rat, an animal model of type 2 diabetes. An enhanced proliferative response to 10% fetal bovine serum(FBS) and superoxide generation with an increased NAD(P)H oxidase activity were observed in diabetic(OLETF) VSMC. Both the enhanced proliferation and superoxide generation in diabetic VSMC were significantly attenuated by AG490, JAK2 inhibitor, and PP2, Src kinase inhibitor. Tyrosine phosphorylation of proteins in diabetic VSMC, especially JAK2, was increased compared to control VSMC. Furthermore, the enhanced NAD(P)H oxidase activity in diabetic VSMC was significantly attenuated by AG490 in a dose-dependent manner. Together, these results indicate that the signal pathway which leads to diabetes-associated activation of Src kinase/JAK is critically involved in the diabetic VSMC proliferation through NAD(P)H oxidase activation and superoxide generation.

• Nutrition Research and Practice
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• v.17 no.5
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• pp.844-854
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• 2023

BACKGROUND/OBJECTIVES: Fucoidan, a polysaccharide content in brown algae, has been reported to inhibit the growth of cancer cells. The present study aimed to investigate the suppression effects of fucoidan on A549 non-small cell lung cancer cells migration. MATERIALS/METHODS: The anti-migratory activity of fucoidan in A549 cells was examined by wound healing assay and phalloidin-rhodamine staining in response to fucoidan (0-100 ㎍/mL) treatment for 48 h. Western blot analysis was performed to clarify the protein expressions relevant to migratory activity. RESULTS: Fucoidan (25-100 ㎍/mL) significantly suppressed A549 cells migration together with reduced the intensity of phalloidin-rhodamine which detect filopodia and lamellipodia protrusions at 48 h of treatment. The protein expression indicated that fucoidan significantly suppressed the phosphorylation of focal adhesion kinase (FAK), Src, and extracellular signal-related kinase (ERK). In addition, the phosphorylation of p38 in A549 cells was found to be increased. CONCLUSIONS: Our data conclude that fucoidan exhibits anti-migratory activities against lung cancer A549 cells mediated by inhibiting ERK1/2 and FAK-Src pathway.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.4
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• pp.56-73
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• 2007

Purpose: 이 실험은 골다공증의 치료약물로 자하거의 골질재흡수 억제효과를 검토하기 위하여 설계되었다. Methods: 자하거의 골질재흡수 효과를 확인하기 위하여 생쥐의 두개골 골모세포를 이용하여 Cyclooxygenase-1(COX-1), COX-2, $TGF-{\beta}$, $L-1{\beta}$, $TNF-{\alpha}$, IL-6, prostaglandin E2등의 활성화 정도를 측정하였으며, 골조직의 미세구조적 변화를 확인하였다. Results: 자하거는 $IL-1{\beta}$, $TNF-{\alpha}$, IL-6 또는 그 세가지의 조합에 의하여 유발된 PGE2의 생성 뿐만 아니라 COX-2 mRNA 수치도 감소시켰으나 COX-1 mRNA 수치에는 영향을 주지 않았다. 이로써 자하거는 시험관내에서 그리고 생체내에서 펩티드의 인산화를 억제함으로써 골의 재흡수를 저해하였다. 그리고 자하거는 생쥐에서 $IL-1{\beta}$에 의해 유발된 고칼슘혈증을 감소시켰고, 골의 재흡수를 저해하는 경로를 통하여 골에 대한 보호효과를 보여줌으로써 조기에 난소 절제한 쥐에서 골질감소와 미세구조적 변화를 부분적으로 방지하였다. 이러한 결과는 PGE2 생성에 대한 $IL-1{\beta}$, $TNF-{\alpha}$, IL-6사이의 상승효과는 COX-2의 유전자 발현이 증가한 결과이며 이러한 tyrosine kinase가 생쥐의 두개골 골모세포에서 COX-2의 신호전달에 관계한다는 것을 보여준다. Conclusion: 자하거가 생쥐의 두개골 골모세포에서 여러 신호전달물질의 활성화를 통하여 골질재흡수를 저해하는 특성을 확인함으로써 앞으로 골다공증의 예방과 치료에 대한 추가적인 임상연구가 필요할 것으로 사료된다.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.15 no.1
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• pp.343-355
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• 2002

Background : Although Samsoeum has been used frequently on allergic rhinitis, but there isn't any experimental research for that. Objective : This study was performed to investigate the anti-allergic effects of Samsoeum and Samsoeumgamibang. Materials and Methods : Katayama's method was used to observe the vascular permeability response induced by serotonin and histamine. Muller's method was used to observe the contact dermatitis response induced by picryl chloride. Miller's method was used to observe the delayed type hypersensitivity response to SRC. Results : 1. In the vascular permeability response to intradermal injection of serotonin, 2,600mg/kg, p.o. group of Samsoeum(蔘蘇飮), 1,300 and 2,600mg/kg, p.o. group of Samsoeumgamibang(蔘蘇飮加味方) showed significant inhibitory effects on the leakage of Evan's blue solution. 2. In the vascular permeability response to intradermal injection of histamine, 1,300 and 2,600mg/kg, p.o. group of Samsoeum, also 1,300 and 2,600mg/kg, p.o. group Samsoeumgamibang showed significant inhibitory effects on the leakage of Evan's blue solution. 3. In the contact dermatitis response induced by picryl chloride, 1,300 and 2,600mg/kg, p.o. group of Samsoeum, 1,300 and 2,600mg/kg, p.o. group of Samsoeumgamibang showed significant inhibitory effects on ear swelling formation. 4. In the delayed type hypersensitivity response to SRC. 2,600mg/kg, p.o. group of Samsoeum and 2,600mg/kg, p.o. group of Samsoeumgamibang showed significant inhibitory effects on foot swelling. Conclusion : This study shows that Samsoeum and Samsoeumgamibang may have anti-allergic effects. So Samsoeum and Samsoeurngamibang can be helpful to treat allergic rhinitis.