• Title/Summary/Keyword: Proliferation control

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A Study for the Mechanism of Abnormal Proliferation in Vascular Endothelial Cells using Inhibitors to the Signal Transduction Pathway (신호전달 경로의 저해제를 이용한 혈관 내피세포의 비정상적인 증식 기전에 대한 연구)

  • Bae, Yong Chan;Park, Suk Young;Nam, Su Bong;Herh, Jae Young;Kang, Young Seok
    • Archives of Plastic Surgery
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    • v.33 no.1
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    • pp.5-12
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    • 2006
  • Protein tyrosine kinase(PTK), protein kinase C(PKC), oxidase, as a mediator, take a significant role in signal transduction pathway of angiogenesis. The authors utilized the inhibitors, targeting the formation of three co-enzyme in signal transduction pathway in order to quantify the suppression of abnormal vascular endothelial cell proliferation induced by DMH, to compare the level suppression in each up-regulated growth factors, CTGF, CYR61, $ITG{\beta}1$, FHL2, and to identify the relationship between abnormal cell proliferation and signal transduction pathway. Five groups were established; Control group, Group of DMH, Group of DMH-mixed Herbimycin, inhibitor of protein tyrosine kinase, Group of DMH-mixed Calphostin C, inhibitor of protein kinase C, Group Of Dmh-Mixed 10U Catalase, Inhibitor Of oxidase. The rise of vascular endothelial cell was compared by MTT assay, and four growth factors were analysed with RT-PCR method, at pre-administration, 4, 8, 12, and 24 hours after administration. In comparison of abnormal proliferation of vascular endothelial cell induced by DMH, suppression was noticed in Herbimycin and Calphostin C group, and Calphostin C group revealed higher suppression effect. Nevertheless, Catalase group did not have any suppression. In manifestation of four growth factors, Herbimycin and Calphostin C group presented similar manifestation with control group, except in $ITG{\beta}$. Catalse group had similar manifestation with DMH group in all four growth factors. Abnormal proliferation of vascular endothelial cell induced by DMH have a direct relationship with PTK and PKC, more specifically to PKC. Oxidase was confirmed not to have any relevance.

Effects of Ginsenosides on $pp60^{c-src}$ Kinase, Intracellular Calcium and Cell Proliferation in NIH 373 Cells

  • Hong, Hee-Youn;Yoo, Gyurng-Soo;Choi, Jung-Kap
    • Journal of Ginseng Research
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    • v.22 no.2
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    • pp.126-132
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    • 1998
  • In the present study, we examined effects of ginseng saponins (ginsenosides) on pp60c-src protein tyrosine kinase (PTK) activity, intracellular calcium concentration ([$Ca^{2+}$]i), and cell proliferation in NIH3T3 cells. Eight different ginsenosides [ginsenoside-Rb1 (G-$Rb_1$), -$Rb_2$, -Rc, -Rd, -Re, -Rf, -$Rg_1$, -$Rg_2$) and ginseng total saponin (GTS) were used for these experiments. All ginsenosides and GTS tested stimulated the activation of $pp60^{c-src}$ kinase, and especially G-$Rb_1$,-Rd,-$Rg_1$, and -$Rg_1$ showed a higher stimulatory effect than others at 16.7 $\mu\textrm{g}$/ml of ginsenosides with a 18 hr-incubation, increasing the activity by 4.5, 3.5, 3.5, and 3.0-fold, respectively, over that of untreated control. In addition, both G-Rd and -$Rg_2$)Rg2 increased ($Ca^{2+}$), to 202 and 334 nM, respectively, about 2-3-fold above the basal level within 7min at 250 $\mu\textrm{g}$/yml of ginsenosides. The increases of ($Ca^{2+}$), were eliminated by Pretreatment of EGTA, an extracellular calcium chelator, suggtasting that they result from an influx of calcium ion from extracellular medium rather than an efflux from intracellular calcium store, endoplasmic reticulum (ER). All ginsenosides studied enhanced cell proliferation to 1.2-1.4-fold over that of untreated control at 5~250 $\mu\textrm{g}$/ml of concentrations. Interestingly the promotion of cell proliferation by ginsenosides corresponded with the activation of c-src kinase, which is an early step in the mitogenic signaling cascade. Taken together, we suggest that some ginsenosides may lead to cellProliferation via the activation of cellular signal transduction Pathway involving $pp60^{c-src}$ kinase.

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Effects of Chitosan on Human Gingival Fibroblasts in Vitro (키토산이 치은섬유아세포에 미치는 영향)

  • Kim, Ok-Su;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.235-247
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    • 2002
  • The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of human gingival fibroblasts. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2, and 2 mg/ml. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized nodules was evaluated after 21 days of culture. The results were as follows : The morphology of cells on the chitosan-coated well was round or spheric. Round cells were aggregated since 6 hours of culture and showed nodule-like appearance after 24 hours of culture and did not achieved confluency at 7 days. The attachment of gingival fibroblasts was inhibited by chitosan coating with a tendency of dose dependent pattern. But, cellular activity of unit cell was higher than control. The proliferation of gingival fibroblasts was inhibited by chitosan coating at 2 mg/ml(P<0.01), while the cell proliferation at 0.02, 0.2 $mg/m{\ell}$ was comparable to the control well. Total alkaline phosphatase activity was inhibited by chitosan coating and decreased in the course of time. While ALP activity of unit cell was the highest at 2mg/ml after 4 days of culture. Finally, gingival fibroblasts produced the mineralized nodule at 2 mg/ml. In summary, the attachment, proliferation, and alkaline phosphatase activity of gingival fibroblasts were influenced differently by the concentration of coated chitosan. From this study, it could be used as the matrix of tissue engineering for gingiva without inhibition on proliferation of gingival fibroblasts using chitosan at the optimal concentration (0.02mg/ml).

The Effect of decalcified Root Surface as PDGF Carrier (PDGF 함유매개체로서 탈회된 치근면의 효과)

  • Woo, Hyo-Sang;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.889-905
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    • 1996
  • It is known that growth factors function as potent biologic mediators regulating numerous activities of wound healing via cell proliferation, migration and extracellular matrix formation and they also promote periodontal regeneration. But, method of growth factor application is controversial yet. So purpose of this study is to evaluate the effect of demineralized root surface as one of method of growth factor application. The ginigival fibroblasts were primary cultured and fifth or sixth subpassages were used in these experiments. In first experiment, root surface blocks demineralized with 100mg/ml tetracycline for 5 minutes and pH 1 citric acid for 3 minutes(experimental groups) and nonteminerilized root surface blocks (control groups) were placed in 100ng/ml PDGF-BB for 5 minutes. Then the cells were seeded on each root surface blocks and cultured for 6, 24, 48, 72 hours. In second experiment, root surface blocks deminerilized with tetracycline and citric acid and nondemineralized root surface blocks were placed in 200ng/ml PDGF-BB for 5 minutes and another non-demineralized root surfcae blocks were placed in DMEM without PDGF-BB. At 1, 2, 4, 6, 8 days, the cells were seeded in 24-well plate and using of each eluent, cultured for 72 hours. The results of the four determinants were presented as mean and S.D.. The results were as follows : The attachment and proliferation of human gingival fibroblast on root surface were more increased when PDGF-BB was applicated on root surfrace demineralized with tetracycline or citric acid than non-demineralized root surface. And, in comparision tetracycline with citric acid, there were more attachment and proliferation of human gingival fibroblast on root surface demineralized with tetracycline than citric acid, and proliferation of human gingival fibroblast on demineralized root surface was increased time dependently 1 day to 3 days. In second experiment using eluent, proliferation of human gingival fibroblast was more increased to 6 days when human gingival fibroblast was cultured in eluent that PDGF-BB was applicated on demineralized root surface than two control groups, and degree of proliferation was decreased time dependently 1 day to 6 days. Proliferation of human gingival fibroblast cultured in eluent without PDGF-BB was constant 1 day to 6 days. After 6 days, degree of proliferation of human gingival fibroblast was similar in four groups. This means that release duration of PDGF-BB from demineralized root surface is 6 days. And in comparision tetracycline with citric acid, there was more proliferation of human gingival fibroblast in tetracycline-treated group than citric acid. In conclusion, demineralized root surface as primary site for PDGF-BB application, especially demineralized with tetracycline has important roles in attachment and proliferation of human gingival fibroblast, and may be useful clinical applications in periodontal regenerative procedures.

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Effect of Geraniol on the Proliferation of L1210 Cells and ICR Mouse Macrophages, and the Activities of Superoxide Dismutase (SOD) and Inducible Nitric Oxide Synthase ( iNOS) Activities (Geraniol이 L1210 세포와 ICR 생쥐 대식세포의 증식,Superoxide Dismutase(SOD)와 Inducible Nitric Oxide Synthase(iNOS) 효소활성에 미치는 영향)

  • Kim, Ji-Yeon;Park, Sie-Won
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.309-316
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    • 2004
  • The present investigation was undertaken to find out the anticancer activity of monoterpene compounds. Monoterpenes showed generally the inhibitory effect on the proliferation o f L1210 cancer cells (cytotoxicity). Geraniol was found to exibit the most potent cytotoxic effect on L1210 cells with an IC50 values of $0.67{\mu}g/ml$. On the other hand, geraniol proved to be capable of stimulating the macrophage proliferation (135% of control). When the life prolonging activity of geraniol by daily oral administration of 0.1~10${\mu}g/10{\mu}l/20$ g body weight to Sarcoma 180 bearing ICR mouse was examined, there was also a significant elevation of survival (best result of 134% of control). The contradictory effects of geraniol on the proliferation of L1210 cells and macrophages proved to be accompanied by the coincident alterations of RNS (reactive nitrogen species) related enzymes activities such as inducible nitric oxide synthase (Inos) in macrophages and ROS (reactive oxygen species) related enzymes activities such as superoxide dismutase (SOD) in L1210 cells, respectively.

Analysis of the Biological Function of ELDF15 Using an Antisense Recombinant Expression Vector

  • Liu, Yan;Wang, Long;Wang, Zi-Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.21
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    • pp.9131-9136
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    • 2014
  • ELDF15, homologous with AT2 receptor-interaction protein 1 (ATIP1), may play an important role in cell differentiation, proliferation, and carcinogenesis. We aimed to understand the biological function of ELDF15 via construction and transfection of a recombinant expression vector containing antisense ELDF15. Recombinant expression vectors were successfully constructed and transfected into K562 cells. A stable transfectant, known as pXJ41-asELDF15, stably produced antisense ELDF15. Compared with K562 and K562-zeo cells, K562-pXJ41-asELDF15 cells showed inhibition of cell proliferation. RT-PCR analysis showed that the expression and protein level of ELDF15 decreased significantly in K562 cells transfected with pXJ41-asELDF15. Expression of hemoglobin increased in K562 cells transfected with pXJ41-asELDF15 by benzidine staining. increases NBT reduction activity in K562 cells transfected with pXJ41-asELDF15.Colony forming efficiency in two-layer soft agar was clearly inhibited as assessed by electron microscopy. These results suggest that ELDF15 plays a potential role in cell differentiation, proliferation and carcinogenesis.

Anticomplementary Activity and Immune-Stimulating Effect of the Extracts from Barley (Hordeum Vulgare) (보리(Hordeum vulgare)추출물의 항보체 및 면역증강 효과)

  • 김유영;구성자
    • Korean journal of food and cookery science
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    • v.13 no.5
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    • pp.661-668
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    • 1997
  • By using several solvents, barley extracts containing the anticomplementary activities in classical pathway were prepared (250 $\mu\textrm{g}$/ml): methanol (83.1%), ethanol (71.9%), water extract (25.4%), M-1 (250 $\mu\textrm{g}$/ml), and the soluble part of methanol extract which showed the highest activity (83.4%) and the yield. Anticomplementary activity of methanol extract as well as protease digestion in classical pathway showed 82.4% and 78.4% in the concentration of 250 $\mu\textrm{g}$/ml, respectively. It was found that protein was not involved in anticomplementary activity in the classical pathway and the methanol extract made an impact on classical pathway, but not on alternative pathway. For the immune-stimulating effect, the T cell proliferation effect of the protease digestion displayed little effect irrespective of the dose. In addition, the T cell proliferation effect of methanol extract showed 13-fold higher proliferation effect compared with positive control. It was revealed that the substance containing protein serves as an important factor for the immune proliferation. Therefore, the anticomplementary activity ${\beta}$-glucan in classical pathway and alternative pathway displayed the lowest activity, showing 2.2%, 22.3% respectively. However, the immune-stimulating effect of ${\beta}$-glucan showed the T cell stimulating effect 13 times higher than positive control.

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Effects of Ubiquitin-conjugating Enzyme 2C on Invasion, Proliferation and Cell Cycling of Lung Cancer Cells

  • Tang, Xiao-Kui;Wang, Ke-Jian;Tang, Yu-Kui;Chen, Li
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.3005-3009
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    • 2014
  • The aims of this study were to investigate the influence of ubiquitin- conjugating enzyme E2C (UBE2C) on biological behavior of lung cancer cells. Using MTT, flow cytometry and invasion assays, we detected UBE2C expression and evaluated its biological properties in these cells, including effects on proliferation, the cell cycle profile and invasive capability. Compared with control cells, the UBE2C transfected cells demonstrated increased cellular proliferation (p<0.05). UBE2C transfected cells also had a lower percentage in G1 phase and a higher percentage in S phase (p<0.05). Importantly, the UBE2C transfected cells had a notable enhancement of cell numbers penetrating the basement membrane compared with the control group (p<0.05). Ectopic up-regulation UBE2C promoted the growth of lung cancer cells in vivo. Furthermore, we found UBE2C increased the expression of cyclin D1 and MMP-2. These results show UBE2C may represent a potential therapeutic target for lung cancer.

Evaluation of Skin Sensitization Potential of Skin Whitening Agents by Local Lymph Node Assay (국소 림프절 시험법(Local lymph node assay)을 이용한 피부미백화장품 원료의 피부감작성 연구)

  • 이종권;박승희;박재현;김형수;엄준호;남기택;한범석;장동덕;최광식
    • Toxicological Research
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    • v.19 no.4
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    • pp.285-291
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    • 2003
  • The use of skin whitening agents has been recently increased in various kinds of cosmetic products, although there were some reports that whitening agents might cause allergic contact dermatitis. A murine local lymph node assay (LLNA) has been developed as an alternative to guinea pigs for contact sensitization potential. This study was carried out to investigate the skin sensitization potential of three whitening agents, arbutin, azelaic acid, and kojic acid, by LLNA using a non-radiois-topic endpoint. Female Balb/c mice were exposed topically to a weak allergen, $\alpha$-hexylcinnamalde-hyde (HCA), and three whitening agents following LLNA protocol. Lymph node (LN) weight and cell proliferation in ears and auricular lymph node using bromodeoxyuridine (BrdU) immunohistochemistry were evaluated. LN weights were significantly increased at the HCA group compared to the vehicle control. A weak allergen, HCA elicited 3-fold or greater increase in cell proliferation of lymph nodes as well as increase in cell proliferation of ear as measured by BrdU immunohistochemistry. However, in the case of skin whitening agent groups, there were no significant changes in LN weight and cell proliferation in the ear and lymph node of mice treated with 5, 10 and 20% of three whitening agents compared to the vehicle control. These results show that these three skin whitening agents may not have contact sensitization potentials at tested concentrations in Balb/c mice by LLNA.

Stimulatory Effects of Ginsenoside-Rg1 on p56lck Kinase and Cell Proliferation in Jurkat T Cells (Jurkat T 세포에서 Ginsenoside-Rg1이 p561ck Kinase 활성과 세포증식에 미치는 영향)

  • Hong, Hee-Youn;Na, Do-Seong;Kwon, Tae-Ik;Choi, Jung-Kap;Yoo, Gyurng-Soo
    • Journal of Ginseng Research
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    • v.19 no.2
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    • pp.117-121
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    • 1995
  • We studied the effects of ginsenoside-$Rg_1$ (G-$Rg_1$) extracted from Panax ginseng C.A. Meyer on $p56^{kk}$ kinase and cell proliferation in Jurkat T cells. $p56^{kk}$ was maximally activated within 5 min after the treatment of 16.7 $\mu\textrm{g}$/ml of G-$Rg_1$ increasing the activity by 1.2-2 times relative to untreated control, thereafter its activity was gradually decreased to the level of untreated control. The action of EGTA on the kinase was altered by the addition of G-$Rg_1$, accompanying the band shift of $p56^{kk}$ to $p60^{kk}$. In addition, G-$Rg_1$promoted cell proliferation in a concentration-dependent manner. These results suggest that G-$Rg_1$ may be involved in T cell receptor-CD3 (TCR) signaling via the activation of $p56^{kk}$ and the chance of cellular calcium concentration.

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