• 생약학회지
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• 제35권1호통권136호
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• pp.62-79
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• 2004

Cell-scattering is a phenotypic change easily observed in most epithelial cells treated with Hepatocyte Growth Factor /scatter Factor (HGF/SF) or phorbol esters (PKC-activators). Recent studies have shown the possibilities to use as therapeutic materials of HGF/SF or non tumor promoting phorbol esters for liver disease, cancer and AIDS. In this study, we tested a cell-scattering activity of 534 methanol extracts from plants inhabiting in Korean peninsula using the phenotype-based assay system. Nine Active extracts were detected : Daphne genkwa, Daphne kiusiana, and Aleurites fordii showed high activity (+++), Euphorbia sieboldiana and Rhodotypos scandens showed medium activity (++), Sambucus sieboldiana var. pendula, Catalpa bignonioides, Sambucus sieboldiana and Lycoris squamigera showed low activity (+). Furthermore, the effects of these active materials in the culture cells were investigated with biochemical studies.

• International Journal of Oral Biology
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• 제39권1호
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• pp.41-47
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• 2014

Streptococcus mutans (S. mutans) is a facultative anaerobic bacterium mainly found in the oral cavity and is known to contribute to tooth decay and gingivitis. Recent studies on intestinal microbiota have revealed that microorganisms forming a biofilm play important roles in maintaining tissue homeostasis through their own metabolism. However, the physiological roles of oral microorganisms such as S. mutans are still unclear. In our current study, we identified that constituents released from S. mutans (CR) reduce arecoline-mediated cytotoxicity without producing toxic effects themselves. Arecoline, as a major alkaloid of areca nut, is known to mediate cytotoxicity on oral epithelial cells and induces a sustained intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) increase that is cytotoxic. The exposure of human gingival fibroblast (HGF) cells to CR not only inhibited the sustained $[Ca^{2+}]_i$ increase but also the initial $[Ca^{2+}]_i$ elevation. In contrast, CR had no effects on the gene regulation mediated by arecoline. These results demonstrate that S. mutans has physiological role in reducing cytotoxicity in HGF cells and may be considered a novel pharmaceutical candidate.

• Journal of Periodontal and Implant Science
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• 제30권4호
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• pp.895-913
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• 2000

It is becoming increasingly clear that human gingival fibroblasts(HGF) may play a role in regulating immune responsiveness in inflammatory periodontal lesions. Stimulation of HGF with locally-secreted T cell cytokine $IFN_{\gamma}$ induces human leukocyte antigen class II(HLA II) expression on HGF, which is one of the characteristic feature of professional antigen presenting cells(pAPC). However, $IFN_{\gamma}$-treated HGF and other nonprofessional antigen presenting cells(npAPC) are known to be ineffective or less effective antigen presenter to resting T cells. This study, therefore, was undertaken in an effort to elucidate the differences in expression of cell surface molecules between npAPC in periodontal tissues, such as HGF and periodontal ligament fibroblasts(PDLF), and pAPC such as monocytes/macrophages. Using flow cytometry, the levels of cell surface expression of HLA-D, ICAM-1, LFA-3, and B7-1, which are involved in antigen presentation, were determined in HGF, PDLF and human myelomonocytic cell line THP-1. $IFN_{\gamma}$ clearly induced HLA-D expression on both of fibroblasts and monocytes dose dependently. However, expression level on monocytes were 4 to 5 times higher than that on fibroblasts, and induction rate was faster in monocytes than in fibroblasts. The levels of ICAM-1 expression on fibroblasts and monocytes were enhanced by $IFN_{\gamma}$ in a dose dependent manner. On the other hand, the expression of LFA-3 molecule, which could be detected in fibroblasts and monocytes without cytokine stimulation, was no more enhanced by addition of $IFN_{\gamma}$. B7-1, important costimulatory molecule in T cell activation and proliferation, was not detected on both of fibroblasts and monocytes even when stimulated with $IFN_{\gamma}$, except on monocytes fully differentiated by pretreatment of PMA and treated by $IFN_{\gamma}$. These results suggest that delayed expression of HLA-D and absence of B7-1 on $IFN_{\gamma}$ - treated fibroblasts may at least in part be involved in the ineffectiveness of fibroblasts as primary APC. And it is postulated that although periodontal fibroblasts may not serve as primary APC in normal periodontium, sustained expression of HLA II on ubiquitous fibroblasts in inflammatory lesions may perpetuate immune responses and produce chronic inflammation and tissue injury.

• Parasites, Hosts and Diseases
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• 제48권4호
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• pp.291-295
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• 2010

The onset, severity, and ultimate outcome of malaria infection are influenced by parasite-expressed virulence factors and individual host responses to these determinants, In both humans and mice, liver injury is involved after parasite entry, which persists until the erythrocyte stage after infection with the fatal strain Plasmodium falciparum (Pf), Hepatocyte growth factor (HGF) has strong anti-apoptotic effects in various kinds of cells, and also has diverse metabolic functions. In this work, Pf-subtilisin-like protease 2 (Pf-Sub2) 5' untranslated region (UTR) was analyzed and its transcriptional activity was estimated by luciferase expression. Fourteen TATA boxes were observed but only one Oct-1 and c-Myb were done. In addition, host HGF interaction with Pf-Sub2 was evaluated by co-transfection of HGF- and Pf-Sub2-cloned vector. Interestingly, -1,422/+12 UTR exhibited the strongest luciferase activity but -329 to + 12 UTR did not exhibit luciferase activity. Moreover, as compared with the control of unexpressed HGF, the HGF protein suppressed luciferase expression driven by the 5' untranslated region of the Pf-Sub2 promoter. Taken together, it is suggested that HGF controls and interacts with the promoter region of the Pf-Sub2 gene.

• 치과방사선
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• 제29권2호
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• pp.435-449
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• 1999

Purpose : The study was aimed to detect the differences in the cell viability and the apoptosis induction after irradiation on normal and tumorigenic cells. Materials and Methods : The study. that was generated for two human normal cells(RHEK, HGF-l) and two human tumor cells(KB. HT-1080). was tested using MTT assay at 1 day and 3 day after irradiation and TUNEL assay under confocal laser scanning microscope at 1 day after irradiation. Single irradiation of 0.5. 1, 2. 4. and 8Gy were applied to the cells. The two fractions of 1. 2. 4. and 8Gy were separated with a 4-hour time interval. The irradiation was done with 5.38Gy/min dose rate using Cs-137 irradiator at room temperature. Results and Conclusions : 1. In 3-day group. the cell viability of HGF-1 cell was significantly decreased at 2. 4 and 8Gy irradiation, the cell viability of KB cell was significantly decreased at 8Gy irradiation and the cell viability of HT-I080 cell was significantly decreased at 4 and 8Gy irradiation. 2. There was significant difference between RHEK and KB cell line in the cell viability of 3-day group at 8Gy irradiation. There was significant difference between RHEK and HGF-1 cell line in the cell viability of 3-day group at 4 and 8Gy irradiation. 3. There was a significantly decreased cell viability in 3-day group than those in 1-day group at 2. 4 and 8Gy on HGF-1 cell. at 4 and 8Gy on HT-I080 cell. at 8Gy on KB cell. 4. We could detect DNA fragmented cells only on KB cell. Number of apoptotic cells of KB cell was significantly increased at 4 and 8Gy irradiation. However, there was no correlation between cell viability and apoptosis. 5. On all 4 cell lines, there were no differences between single and split irradiation method in cell viability and apoptosis.

• Journal of Microbiology and Biotechnology
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• 제23권9호
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• pp.1327-1338
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• 2013

The humanized anti-hepatocyte growth factor (HGF) monoclonal antibody (mAb) YYB-101 is a promising therapeutic candidate for treating various cancers. In this study, we developed a bioprocess for large-scale production of YYB-101 and evaluated its therapeutic potential for tumor treatment using a xenograft mouse model. By screening diverse chemically defined basal media formulations and by assessing the effects of various feed supplements and feeding schedules on cell growth and antibody production, we established an optimal medium and feeding method to produce 757 mg/l of YYB-101 in flask cultures, representing a 7.5-fold increase in titer compared with that obtained under non-optimized conditions. The optimal dissolved oxygen concentration for antibody production was 70% $pO_2$. A pH shift from 7.2 to 7.0, rather than controlled pH of either 7.0 or 7.2, resulted in productivity improvement in 5 L and 200 L bioreactors, yielding 737 and 830 mg/ml of YYB-101, respectively. The YYB-101 mAb highly purified by affinity chromatography using a Protein A column and two-step ion exchange chromatography effectively neutralized HGF in a cell-based assay and showed potent tumor suppression activity in a mouse xenograft model established with human glioblastoma cells.

• 한국자원식물학회지
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• 제35권5호
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• pp.565-573
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• 2022

치주조직에 존재하는 주요한 세포의 한 형태인 인체 치은섬유아세포는 다양한 구강유해세균으로부터 염증이 유발되어지며, 그중 대표적으로 치주염 원인균인 P. gingivalis의 내독소인 LPS-PG로부터 염증성 자극에 반응하여 다양한 염증매개 물질을 분비한다. 본 연구에서는 치주염을 일으키는 주요한 원인균 중 하나인 P. gingivalis로 부터 분리한 LPS-PG를 이용하여 인체 치은섬유아세포주인 HGF-1 세포에 염증을 유도한 후 LRE에 대한 항염증 및 항산화 효과를 분석하였다. 실험 결과, LRE는 LPS-PG 유도에 따라 iNOS에 의한 NO 생성과 COX-2에 의한 PGE₂와 같은 염증 매개 인자의 발현 및 생성 억제와 함께 염증성 싸이토카인(TNF-α, IL-1β및 IL-6)의 생성 또한 억제하였다. 신호전달계에서 염증성 전사인자의 발현 경로를 확인하기 위하여 TLR4/Myd88/NF-κB의 활성을 확인한 결과, LRE 처리에 따라 농도 의존적으로 억제되는 것을 확인하였다. 또한 산화 환원 효소로 항염증효과를 나타내는 것으로 알려진2상 효소 중 하나인 NQO-1과 이의 전사인자인 Nrf2를 분석 한 결과 LRE 처리에 의해 효소의 활성이 높아지는 것을 확인할 수 있었다. 결론적으로 LRE는 TLR4/Myd88/NF-κB 신호전달 경로를 억제하고 NQO1/Nrf2 활성을 유도함으로써 HGF-1 세포에서 LPS-PG에 의해 유도된 염증을 억제하는 것으로 사료되며, 향후 LRE는 식·의약품 소재 개발에서 치주질환 개선의 가능성이 있는 후보물질이 될 수 있을 것으로 사료된다.

• 한국임상수의학회:학술대회논문집
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• 한국임상수의학회 2009년도 추계학술대회
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• pp.265-265
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• 2009

• Journal of Microbiology and Biotechnology
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• 제23권8호
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• pp.1176-1184
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• 2013

Anti-hepatocyte growth factor (anti-HGF) monoclonal antibodies (mAbs) are potential therapeutics against various cancers. Screening for high-producer clones is a time-consuming and complex process and is a major hurdle in the development of therapeutic mAbs. Here, we describe an efficient approach that allows the selection of high-producer Chinese hamster ovary (CHO) cell lines producing the novel anti-HGF mAb SFN68, which was generated previously by immunizing HGF bound to its receptor c-Met. We selected an SFN68-producing parental cell line via transfection of the dihydrofolate reductase-deficient CHO cell line DG44, which was preadapted to serum-free suspension culture, with an SFN68-expression vector. Subsequent gene amplification via multiple passages of the parental cell line in a methotrexate-containing medium over 4 weeks, followed by clonal isolation, enabled us to isolate two cell lines, 2F7 and 2H4, with 3-fold higher specific productivity. We also screened 72 different media formulated with diverse feed and basal media to develop a suboptimized medium. In the established suboptimized medium, the highest anti-HGF mAb yields of the 2F7 and 2H4 clones were 842 and 861 mg/l, respectively, which were about 10.5-fold higher than that of the parental cell line in a non-optimized basal medium. The selected CHO cell lines secreting high titers of SFN68 would be useful for the production of sufficient amounts of antibodies for efficacy evaluation in preclinical and early clinical studies.

• 한국융합학회논문지
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• 제9권12호
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• pp.81-88
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• 2018

사람치은섬유모세포(human gingival fibroblast, HGF)는 치은조직에 존재하는 주요한 세포의 형태 중 하나로 외부 자극에 반응하여 다양한 염증대사물질을 생산한다. 본 연구에서는 돌김에탄올추출물(PYEE)이 Porphyromonas gingivalis로부터 분리한 lipopolysaccharide로 염증이 유도된 HGF-1 cell에서 항염 효과를 보이는지 분석하고자 하였다. LPS-PG에 의해 과발현된 iNOS와 COX-2는 PYEE의 처리에 의해 농도 의존적으로 발현이 감소되었고, nuclear factor $(NF)-{\kappa}B$ 또한 동일한 양상으로 활성이 억제되었다. 신호전달물질 중 c-Jun $NH_2$-terminal kinase (JNK)의 인산화만이 PYEE에 의해 억제되었다. 그리고 항염 작용에 관여하는 것으로 알려진 2상 효소 중 하나인 NAD(P)H:quinone dehydrogenase (NQO)-1도 분석하였고, 이 효소는 PYEE의 처리에 의해 강하게 발현이 유도가 되었다. 결론적으로 돌김에탄올추출물은 치주질환 에방과 치료를 위한 후보물질로 활용 가능할 것으로 사료된다.