• Title/Summary/Keyword: Small protein

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Reversal of Cisplatin Resistance by Epigallocatechin Gallate Is Mediated by Downregulation of Axl and Tyro 3 Expression in Human Lung Cancer Cells

  • Kim, Kyung-Chan;Lee, ChuHee
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.1
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    • pp.61-66
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    • 2014
  • Lung cancer is still the number one cause of death from cancer worldwide. The clinical effect of platinum-based chemotherapy for non-small cell lung cancer is constrained by the resistance to drug. To overcome chemo-resistance, various modified treatment including combination therapy has been used, but overall survival has not been improved yet. In this study, chemo-resistant lung cancer cells, A549/Cis and H460/Cis, were developed by long-term exposure of cells to cisplatin and the proliferative capability of these resistant cells was verified to be reduced. We found cytotoxic effect of epigallocatechin gallate (EGCG), a major catechin derived from green tea, on both the parental lung cancer cells, A549 and H460, and their cisplatin resistant cells, A549/Cis and H460/Cis. ELISA and Western blot analysis revealed that EGCG was able to increase interlukine-6 (IL-6) production per cell, whereas its downstream effector Signal transducers and activators of transcription 3 (STAT3) phosphorylation was not changed by EGCG, indicating that IL-6/STAT3 axis is not the critical signaling to be inhibited by EGCG. We next found that EGCG suppresses the expression of both Axl and Tyro 3 receptor tyrosine kinases at mRNA and protein level, explaining the cytotoxic effect of EGCG on lung cancer cells, especially, regardless of cisplatin resistance. Taken together, these data suggest that EGCG impedes proliferation of lung cancer cells including their chemo-resistant variants through downregulation of Axl and Tyro 3 expression.

Resveratrol raises in vitro anticancer effects of paclitaxel in NSCLC cell line A549 through COX-2 expression

  • Kong, Fanhua;Zhang, Runqi;Zhao, Xudong;Zheng, Guanlin;Wang, Zhou;Wang, Peng
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.5
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    • pp.465-474
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    • 2017
  • The aim of this study was to determine the raising anticancer effects of resveratrol (Res) on paclitaxel (PA) in non-small cell lung cancer (NSCLC) cell line A549. The $10{\mu}g/ml$ of Res had no effect on human fetal lung fibroblast MRC-5 cells or on A549 cancer cells and the 5 or $10{\mu}g/ml$ of PA also had no effect on MRC-5 normal cells. PA-L ($5{\mu}g/ml$) and PA-H ($10{\mu}g/ml$) had the growth inhibitory effects in NSCLC cell line A549, and Res increased these growth inhibitory effects. By flow cytometry experiment, after Res ($5{\mu}g/ml$)+PA-H ($10{\mu}g/ml$) treatment, the A549 cells showed the most apoptosic cells compared to other group treatments, and after additional treatment with Res, the apoptosic cells of both two PA concentrations were raised. Res+PA could reduce the mRNA and protein expressions of COX-2, and Res+PA could reduce the COX-2 related genes of VEGF, MMP-1, MMP-2, MMP-9, $NF-{\kappa}B$, Bcl-2, BclxL, procollagen I, collagen I, collagen III and CTGF, $TNF-{\alpha}$, $IL-1{\beta}$, iNOS and raise the TIMP-1, TIMP-2, TIMP-3, $I{\kappa}B-{\alpha}$, p53, p21, caspase-3, caspase-8, caspase-9, Bax genes compared to the control cells and the PA treated cells. From these results, it can be suggested that Res could raise the anticancer effects of PA in A549 cells, thus Res might be used as a good sensitizing agent for PA.

Evaluation of Eating Habit and Dietary Intake by Family Type of Seniors Utilizing Social Welfare Center in Seoul (서울시내 10개소 노인 복지관 이용자의 가족 형태에 따른 식습관 분석 및 영양소 섭취량 조사)

  • Kim, Eun-Mee;Song, Yeon-Joo;Kim, Hyun-Sook
    • Journal of Nutrition and Health
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    • v.40 no.8
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    • pp.753-761
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    • 2007
  • The objective of our study was to evaluate the dietary intakes and eating habit for seniors utilizing ten different social welfare centers in Seoul, Korea. The total number of subjects of this study was 543(male: 199, female: 344) of 60 years of age or older. The survey was performed through personal interview questionnaires from May 3rd to June 1st 2006. After assigning subjects to four categories based on family type, single, couple, joint family, and other, these categories were compared with the estimated total nutrient intakes and eating habit. Validity was assessed by comparison with 24-hour dietary recalls and dietary variety score(DVS). The results showed that senior in single family type was far lower than that of couple family type in terms of frequency of meal per day and intake of snacks. Dietary variety score represented that a single female with social security beneficiary showed the lowest variety score compared with other family type. The lunch meal provided by social welfare center turned out to be the most nutritious diet and had significant proportion of daily intakes in essential nutrients such as protein, iron, vitamin A, and vitamin C. Taken together, the family type was a significant factor and the accompaniment is another effector on dietary pattern in the elderly. We strongly suggest that the current lunch program in welfare center should be expanded to breakfast meal, and furthermore the small commodity program is also needed to provide the elderly with meal for social communication.

Fermentation Characteristics of Kimchi Supplemented with Cheese. (치즈를 첨가한 김치의 발효 특성)

  • 배인휴;최성희;최희영
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.415-419
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    • 2002
  • The replacement effects of cheese far salted and fermented fish on growth of lactic acid bacteria, fermentation velocity and sensory characteristics of Kimchi were investigated. In both control and cheese Kimchi, the total viable cell count of lactic acid bacteria was increased rapidly during the initial 2 days of fermentation. From 3 days after preparation, Kimchi added with cheese showed higher number of lactic acid bacteria than control Kimchi. The pH of Kimchi decreased rapidly after a small increase at the first day of fermentation, reaching 4.18-4.33 at the third day of fermentation, and the pH was slightly lower in Kimchi added with cheese than in control. Proximate analysis of Kimchi added with cheese was slightly higher in moisture and lower in crude protein and fat than control Kimchi. Sensory evaluation of the Kimchi fermented for 3 days showed that the Kimchi added with 3 or 5% of cheese had higher scores of appearance, flavor and overall taste than the control Kimchi.

Molecular Interactions of a Replication Initiator Protein, RepA, with the Replication Origin of the Enterococcal Plasmid p703/5

  • Cha, Kyung-Il;Lim, Ki-Hong;Jang, Se-Hwan;Lim, Wang-Jin;Kim, Tae-Hyung;Chang, Hyo-Ihl
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1841-1847
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    • 2007
  • We previously identified the origin of replication of p703/5, a small cryptic plasmid from the KBL703 strain of Enterococcus faecalis. The origin of replication contains putative regulatory cis-elements required for replication and a replication initiator (RepA) gene. The replicon of p703/5 is similar in its structural organization to theta-type plasmids, and RepA is homologous to a family of Rep proteins identified in several plasmids from Gram-positive bacteria. Here, we report molecular interactions between RepA and the replication origin of p703/5. DNase I footprinting using recombinant RepA together with electrophoretic mobility shift assays confirmed the binding of RepA to the replication origin of p703/5 via iterons and an inverted repeat. We also demonstrated the formation of RepA dimers and the different binding of RepA to the iteron and the inverted repeat using gel filtration chromatographic analysis, a chemical crosslinking assay, and electrophoretic mobility shift assays in the presence of guanidine hydrochloride. Our results suggest that RepA plays a regulatory role in the replication of the enterococcal plasmid p703/5 via mechanisms similar to those of typical iteroncarrying theta-type plasmids.

p38 MAPK Participates in Muscle-Specific RING Finger 1-Mediated Atrophy in Cast-Immobilized Rat Gastrocnemius Muscle

  • Kim, Jung-Hwan;Won, Kyung-Jong;Lee, Hwan-Myung;Hwang, Byong-Yong;Bae, Young-Min;Choi, Whan-Soo;Song, Hyuk;Lim, Ki-Won;Lee, Chang-Kwon;Kim, Bo-Kyung
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.6
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    • pp.491-496
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    • 2009
  • Skeletal muscle atrophy is a common phenomenon during the prolonged muscle disuse caused by cast immobilization, extended aging states, bed rest, space flight, or other factors. However, the cellular mechanisms of the atrophic process are poorly understood. In this study, we investigated the involvement of mitogen-activated protein kinase (MAPK) in the expression of muscle-specific RING finger 1 (MuRF1) during atrophy of the rat gastrocnemius muscle. Histological analysis revealed that cast immobilization induced the atrophy of the gastrocnemius muscle, with diminution of muscle weight and cross-sectional area after 14 days. Cast immobilization significantly elevated the expression of MuRF1 and the phosphorylation of p38 MAPK. The starvation of L6 rat skeletal myoblasts under serum-free conditions induced the phosphorylation of p38 MAPK and the characteristics typical of cast-immobilized gastrocnemius muscle. The expression of MuRF1 was also elevated in serum-starved L6 myoblasts, but was significantly attenuated by SB203580, an inhibitor of p38 MAPK. Changes in the sizes of L6 myoblasts in response to starvation were also reversed by their transfection with MuRF1 small interfering RNA or treatment with SB203580. From these results, we suggest that the expression of MuRF1 in cast-immobilized atrophy is regulated by p38 MAPK in rat gastrocnemius muscles.

In vivo and In vitro Metabolism of Recombinant Human Epidermal Growth Factor (DWP401) in Rats (재조합 인간 상피세포성장인자(DWP401)의 흰쥐에서의 in vivo와 in vitro 대사)

  • Koh, Yeo-Wook;Nam, Kouen-Ho;Jung, Ju-Young;Park, Seung-Kook;Yu, Young-Hyo;Kim, Jae-Hwan;Han, Kun;Park, Myung-Hwan;Shim, Chang-Koo
    • YAKHAK HOEJI
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    • v.41 no.3
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    • pp.381-388
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    • 1997
  • Metabolism of DWP401, recombinant juman epidermal growth factor, was examined in vivo and in vitro in rats. When $^{125}I$-labeled DWP401 was administered at a dose of 50 ${\mu}g$/kg by i.v. injection. $^{125}I$-labeled DWP401 was rapidly degraded within 30 minytes above 93%. Thin layer chromatography analysis of urine collected for 24 hr after i.v. administration of $^{125}I$-labeled DWP401 showed ohly one spot on a X-ray film which was considered as diiodo-tyrosine. This result suggests tha $^{125}I$-labeled DWP401 was completely digested into free amino acids without any specific intermediate polypeptides. About 42.1% of the administered iodine was recovered in 24 hr. For in vitro degradation study, $^{125}I$-labeled DWP401 was added to plama and tissue homogenates of rats and incubated at $37^{\circ}C$. Almost 98% of the added radioactivity recovered from the protein fraction of the liver, kidey, small intestine, stomach and spleen decreased rapidly. For examplem the recovery rates of $^{125}I$-labeled DWP401 were 58.6, 63.2, 39.9, 52.9 and 66.8% after 4hrs of incubation in respective organ homogenates.

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Search Space Reduction Techniques in Small Molecular Docking (소분자 도킹에서 탐색공간의 축소 방법)

  • Cho, Seung Joo
    • Journal of Integrative Natural Science
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    • v.3 no.3
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    • pp.143-147
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    • 2010
  • Since it is of great importance to know how a ligand binds to a receptor, there have been a lot of efforts to improve the quality of prediction of docking poses. Earlier efforts were focused on improving search algorithm and scoring function in a docking program resulting in a partial improvement with a lot of variations. Although these are basically very important and essential, more tangible improvements came from the reduction of search space. In a normal docking study, the approximate active site is assumed to be known. After defining active site, scoring functions and search algorithms are used to locate the expected binding pose within this search space. A good search algorithm will sample wisely toward the correct binding pose. By careful study of receptor structure, it was possible to prioritize sub-space in the active site using "receptor-based pharmacophores" or "hot spots". In a sense, these techniques reduce the search space from the beginning. Further improvements were made when the bound ligand structure is available, i.e., the searching could be directed by molecular similarity using ligand information. This could be very helpful to increase the accuracy of binding pose. In addition, if the biological activity data is available, docking program could be improved to the level of being useful in affinity prediction for a series of congeneric ligands. Since the number of co-crystal structures is increasing in protein databank, "Ligand-Guided Docking" to reduce the search space would be more important to improve the accuracy of docking pose prediction and the efficiency of virtual screening. Further improvements in this area would be useful to produce more reliable docking programs.

Resveratrol inhibits cell growth via targeting the Bmi-1 pathway in YD-10B human oral squamous cell carcinoma cells

  • Park, Kyoung-Eun;Ok, Chang Youp;Jang, Hye-Ock;Bae, Moon-Kyoung;Bae, Soo-Kyung
    • International Journal of Oral Biology
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    • v.45 no.3
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    • pp.115-125
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    • 2020
  • Resveratrol has been reported to exert anticancer activity via modulation of multiple pathways and genes. In this study, we examined the effect of resveratrol on YD-10B human oral squamous cell carcinoma cells and its molecular mechanisms of action. We found that resveratrol inhibited the proliferation of YD-10B cells in a dose- and time-dependent manner. The suppressive effect of resveratrol was accompanied by a reduction in Bmi-1 gene expression. We observed that silencing the Bmi-1 gene by small interfering RNA effectively downregulated the levels of GLUT1 mRNA and protein, which were also repressed by resveratrol. Bmi-1 silencing increased the number of YD-10B cells in S-phase arrest by approximately 2.3-fold compared with the control. In conclusion, the results of the present study demonstrate, for the first time, that resveratrol suppresses Bmi-1-mediated GLUT1 expression in human oral squamous cell carcinoma cells and suggest that the specific molecular targeting of Bmi-1 and/or GLUT1 expression can be combined with a chemotherapeutic strategy to improve the response of oral cancer cells to resveratrol.

THE HISTOLOGIC STUDY OF THE GRAFTED hBMP-I FOR IMMEDIATE IMPLANT FIXATION (발치 후 즉시 임플란트 식립시 이식된 hBMP-I의 조직학적 고찰)

  • Lee, Eun-Young;Kim, Kyoung-Won;Choi, Hee-Won;Um, In-Woong;Chung, Ho-Yong
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.4
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    • pp.316-322
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    • 2004
  • A low molecular weight component named bone morphogenetic protein(BMP) chemically isolated from the organic matrix of bone, induce postfetal connective tissue cells surrounding small blood vessels to differentiate into cartilage and bone. The end product of BMP is a spherical ossicle of lamella bone filled with red bone marrow for the functional loading. This is a important point that the graft material is embedded the defect site during the implant surgery. Because present knowledge of the relationship between BMP and bone regeneration arises mainly from studies of induced bone formation in heterotopic sites, it would be helpful to determine whether BMP plays any part in the process of bone healing. The BMPs have been shown to play crucial roles in normal skeletal development as well as bone healing and are able to activate transcription of genes involved in cellular migration, proliferation, and differentiation. The delivery of BMP on matrices has been efficacious in the treatment of defect bone in implant surgery. The purpose of the histologic study was to evaluate the effect of DLB(demineralized lyophilized bone) coated with purified human BMP(hBMP-I) in immediate implant surgery with bony defect to obtain the functional structure of implant asap. The ability of a graft of hBMP-I to accelerate bony defect repair provides a rationale for its use in immediate implant surgery that have large bone defect in edentulous area.