• Title/Summary/Keyword: Adhesion assay

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Antioxidative Effect and Melanogenesis of Nelumbo nucifera Stamen Extract on Cultured Human Skin Melanoma Cells Injured by Hydrogen Peroxide (연꽃수술추출물이 과산화수소로 손상된 배양 인체피부흑색종세포에 대한 항산화효과 및 멜라닌화에 미치는 영향)

  • Kim, Myoung-Seoup;Park, Yun-Jum;Sohn, Young-Woo
    • Korean Journal of Plant Resources
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    • v.23 no.2
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    • pp.145-150
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    • 2010
  • To examine the antioxidative effect and melanogenesis of Nelumbo nucifera stamen (NNS) extract on hydrogen peroxide $H_2O_2$ induced cytotoxicity in cultured human skin melanoma cells (SK-MEL-3), cell adhesion activity (CAA), tyrosinase inhibitory activity and total amount of melanin synthesis were measured by colorimetric assay. In this study, $H_2O_2$ significantly decreased CAA, and $CAA_{50}$ value of $H_2O_2$ was determined at 30 uM. In the antioxidative effect, NNS extract increased cell adhesion activity which was decreased by $H_2O_2$ induced cytotoxicity, and also, tyrosinase activity and total amount of melanin were decreased by NNS extract. These results suggested that $H_2O_2$ was highly toxic on cultured human skin melanoma cells and NNS extract showed the antioxidative and inhibitory effect of melanogenesis by the increased CAA, and the decresed tyrosinase activity and total amount of melanin synthesis.

The Effect of Chrysanthemum morifolium L. Extract on Cultured Neuroglial Cells Damaged by Glucose Oxidase

  • Seo, Young-Mi;Park, Seung-Taeck;Rim, Yo-Sup;Chung, Ok-Bong;Jekal, Seung-Joo
    • Korean Journal of Clinical Laboratory Science
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    • v.43 no.2
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    • pp.75-81
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    • 2011
  • To clarify the oxidative stress of reactive oxygen species (ROS) and the effect of Chrysanthemum morifolium L. (CM) flower extract on the cultured neuroglial cells (C6 glioma) damaged by ROS, cell adhesion effect was measured by colorimetric assay after cultured C6 glioma cells were treated with various concentrations of glucose oxidase (GO) for 5 hours. For the antioxidative effect of CM flower extract, cell adhesion activity (CAA), superoxide dismutase (SOD)-like activity and lactate dehydrogenase (LDH) activity were assessed against GO-induced cytotoxicity on same cultures. In this study, GO remarkably decreased CAA dose-dependently, and the $XTT_{90}$ and $XTT_{50}$ values were measured at 15 mU/mL and 50 mU/mL following the treatment of C6 glioma cells with 5~60 mU/mL of GO. The CM flower extract significantly increased cell adhesion activity damaged by GO-induced cytotoxicity, and it also showed the SOD-like activity and the decrease of LDH activity. From these results, it is suggested that GO was cytotoxic on cultured C6 glioma cells, and CM flower extract showed antioxidative effects as shown by the increased CAA, SOD-like activity and the decrease of LDH activity on GO-induced cytotoxicity on the same cultures.

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Differential Wnt11 Expression Related to Wnt5a in High- and Low-grade Serous Ovarian Cancer: Implications for Migration, Adhesion and Survival

  • Jannesari-Ladani, Farnaz;Hossein, Ghamartaj;Izadi-Mood, Narges
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1489-1495
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    • 2014
  • Wnt is a powerful signaling pathway that plays a crucial role in cell fate determination, survival, proliferation and motility during development, in adult tissues and cancer. The aims of the present study were three fold: i) to assess Wnt11 immunoexpression and its possible relationship with Wnt5a in high- and low-grade human serous ovarian cancer (HGSC and LGSC) specimens; ii) to assess Wnt11 expression levels in Wnt5a overexpressing SKOV-3 cells; iii) to reveal the role of Wnt11 in viability, adhesion, migration and invasion of SKOV-3 cells using recombinant human Wnt11 (rhWnt11). Immunohistochemistry revealed a significant difference in Wnt11 expression between HGSC and LGSC groups (p=0.001). Moreover, a positive correlation was observed between Wnt5a and Wnt11 expression in the HGSC (r=0.713, p=0.001), but not the LGSC group. The expression of Wnt11 was decreased by 35% in Wnt5a overexpressing cells (SKOV-3/Wnt5a) compared to mock controls. Similarly Wnt11 expression levels were decreased by 47% in the presence of exogenous Wnt5a compared to untreated cells. In the presence of rhWnt11, 31% increased cell viability (p<0.001) and 21% increased cell adhesion to matrigel (p<0.01) were observed compared to control. Cell migration was increased by 1.6-fold with rhWnt11 as revealed by transwell migration assay (p<0.001). However, 45% decreased cell invasion was observed in the presence of rhWnt11 compared to control (p<0.01). Our results may suggest that differential Wnt11 immunoexpression in HGSC compared to LGSC could play important roles in serous ovarian cancer progression and may be modulated by Wnt5a expression levels.

Platycodin D Induces Apoptosis, and Inhibits Adhesion, Migration and Invasion in HepG2 Hepatocellular Carcinoma Cells

  • Li, Ting;Xu, Wen-Shan;Wu, Guo-Sheng;Chen, Xiu-Ping;Wang, Yi-Tao;Lu, Jin-Jian
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1745-1749
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    • 2014
  • Background: Platycodin D (PD), a triterpenoid saponin isolated from the Chinese medicinal herb Platycodonis radix, possesses anti-cancer effects in several cancer cell lines. The aim of this study was to evaluate its anticancer activities in hepatocellular carcinoma cells. Materials and Methods: MTT and colony formation assays were performed to evaluate cell proliferation, along with flow cytometry and Western blotting for apoptosis. Cell adhesion was tested by observing cellular morphology under a microscope, while the transwell assay was employed to investigate the cell migration and invasion. Results: PD concentration-dependently inhibited cell proliferation in both HepG2 and Hep3B cells, and significantly suppressed colony formation and induced apoptosis in HepG2 cells. The protein levels of cleaved poly ADP-ribose polymerase (PARP) and Bax were up-regulated while that of survivin was down-regulated after treatment with PD. Moreover, PD not only obviously suppressed the adhesion of HepG2 cells to Matrigel, but also remarkably depressed their migration and invasion induced by 12-O-tetradecanoylphorbol 13-acetate (TPA). Conclusions: PD presents anti-cancer potential in hepatocellular carcinoma cells via inducing apoptosis, and inhibiting cell adhesion, migration and invasion, indicating promising features as a lead compound for anti-cancer agent development.

Suppression of Human Breast Cancer Cell Metastasis by Coptisine in Vitro

  • Li, Jing;Qiu, Dong-Min;Chen, Shao-Hua;Cao, Su-Ping;Xia, Xue-Lan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5747-5751
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    • 2014
  • Background: Coptisine, an isoquinoline alkaloid extracted from Coptidis rhizoma, has many biological activities such as antidiabetic, antimicrobial and antiviral actions. However, whether coptisine exerts anti-cancer metastasis effects remains unknown. Materials and Methods: Effects of coptisine on highly metastatic human breast cancer cell MDA-MB-231 proliferation were evaluated by trypan blue assay and on cell adhesion, migration and invasion by gelatin adhesion, wound-healing and matrigel invasion chamber assays, respectively. Expression of two matrix metalloproteinases (MMPs), MMP-9, MMP-2 and their specific inhibitors tissue inhibitor of metalloproteinase 1 (TIMP-1) and tissue inhibitor of metalloproteinase 2 (TIMP-2) were analyzed by RT-PCR. Results: Coptisine obviously inhibited adhesion to an ECM-coated substrate, wound healing migration, and invasion through the matrigel in MDA-MB-231 breast cancer cells. RT-PCR revealed that coptisine reduced the expression of the ECM degradation-associated gene MMP-9 at the mRNA level, and the expression of TIMP-1 was upregulated in MDA-MB-231 cells, while the expression of MMP-2 and its specific inhibitor TIMP-2 was not affected. Conclusions: Taken together, our data showed that coptisine suppressed adhesion, migration and invasion of MDA-MB-231 breast cancer cells in vitro, the down-regulation of MMP-9 in combination with the increase of TIMP-1 possibly contributing to the anti-metastatic function. Coptisine might be a potential drug candidate for breast cancer therapy.

Anti-inflammatory Effect of Evodia Officinalis $D_{ODE}$ in Mouse Macrophage and Human Vascular Endotherial Cells (마우스 대식세포 및 사람 혈관 내피세포에서 오수유(Evodia officinalis $D_{ODE}$) 메탄올 추출물의 항염증 효과)

  • Yun, Hyun-Jeung;Heo, Sook-Kyoung;Lee, Young-Tae;Park, Won-Hwan;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.23 no.1
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    • pp.29-38
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    • 2008
  • Objectives : Evodia officinalis DODE (EO), an herbal plant, has been widely used in traditional Korean medicine for the treatment of vascular diseases such as hypertension. The crude extract of EO contains phenolic compounds that are effective in protecting liver microsomes, hepatocytes, and erythrocytes against oxidative damage. But EO has been little found to have an anti-inflammatory activity. We investigated anti-inflammatory activity of EO in RAW 264.7 cells and human umbilical vein endothelial cells (HUVECs). Methods : Cytotoxic activity of EO on RAW 264.7 cells was investigated by using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines were measured by ELISA kit. The levels of intracellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression were measured by flow cytometer. Results : EO decreased LPS-induced NO production in RAW 264.7 cells. The inhibitory activity of EO on LPS-induced NO release is probably associated with suppressing TNF-${\alpha}$, IL-6 and MCP-1 formation. These results indicate that EO has potential as an anti-inflammatory agent. Moreover, EO decreased TNF-${\alpha}$-induced IL-8, IL-6 production, and ICAM-1 and VCAM-1 expression in HUVECs. Conclusions : EO inhibits TNF-${\alpha}$-induced inflammation via decreasing cytokines production and adhesion molecules expression. These results indicate that EO has potential as an anti-inflammation and anti-artherosclerosis agent.

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Study on Study on Antitumor Activity of Kamisamchulsamja-tang (가미삼출삼자탕의 항암활성에 관한 연구)

  • Kim Seong Eon;Lee Hyo Jeong;Kim Dong Hee;Song Gyu Yong;Kim Sung Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.4
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    • pp.667-673
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    • 2002
  • The purpose of this study was to investigate the effect of Kamisamchulsamja-tang (KSST) water extract on the antitumor activity. The results were summarized as follows: KSST extract exhibited a weak cytotoxicity against HT1080, A549, SK-OV-3, B16-F10 and SK-MEL-2 cells. KSST extract showed a inhibitoty effect on DNA topoisomerase I from calf thymus in a dose-dependent manner. Also, KSST extract showed antiadhesive effect on HT1080 cells but didn't showed on A549 cells to complex extracellular matrix. In pumonary colonization assay, a number of colonies in the lungs were decreased significantly in KSST treated group as compared with control group. In vitro neovascularization assays, angiogenesis was significantly inhibited in KSST treated group than control group. In CAM assay, KSST extract inhibited angiogenesis significantly at 15㎍/egg concentration as compared with control. From the above results it was concluded that KSST showed antitumor effect through the antimetastatic effect. So it is expected to be clinically helpful on the prevention of metastasis of cancer.

Study on trends of cancer study in TKM and its research strategy in future (한의학계(韓醫學界)의 암연구동향(癌硏究動向)과 연구(硏究) 전략(戰略)에 대한 연구(硏究))

  • Kim, Sung-Hoon
    • The Journal of Korean Medicine
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    • v.19 no.1
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    • pp.470-499
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    • 1998
  • For the purpose of designing more successful cancer research, the strategy for cancer study in the field of Traditional Korean Medicine(TKM) during the 21th century was examined from the analysis of trends on cancer study in traditional korean medicine. The results were summarized as follows: 1. So far cancer research in TKM was chiefly done on cytotoxicity, side-effects by chemotherapy, tumor immunology, apoptosis, survival time with S-180 and pulmonary colonization assay and also clonogenic assay, cell adhesion assay, angiogenesis, cell-differentiation and side-effect by radiotherapy were partly performed. 2. It may be ideal that we should study synergistic effect between constituent drugs of prescriptions, tumor immunology, combined therapy between western and oriental medicines by reducing side-effect by radiotherapy and chemotherapy and antimetastasis according to the characteristics of oriental medicine chiefly and also supplement the studies on molecular biology, gene therapy, angiogensis and signal transduction. 3. We had better do specific-field research in cooperation between oriental medical colleges and Korea Institute of Oriental Medicine(KIOM) as well as study a target cancers such as hepatic cancer, pulmonary cancer and gastric cancer more intensively than all cancers domestically. 4. Our country must keep communication with China having many clinical data, Taiwan chiefly doing the combined tharapy between oriental and western medicines, Japan having done basic study actively on cancer.

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Thrombospondins Mediate the Adhesion of Osteoblast to Extracelluar Matrix

  • Lim, Dong-Jin;Bae, In-Ho;Jeong, Byung-Chul;Kim, Sun-Hun;Park, Bae-Keun;Kang, In-Chul;Lee, Shee-Eun;Song, Sang-Hun;Koh, Jeong-Tae
    • International Journal of Oral Biology
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    • v.33 no.3
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    • pp.105-111
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    • 2008
  • Thrombospondins (TSP-1, TSP-2) are secretory extracellular glycoproteins that are involved in a variety of physiological processes such as tumor cell adhesion, invasion, and metastasis. The present study was undertaken to elucidate the involvement of thrombospondins in the adhesion of osteoblast-like cells using the TSP-1 or TSP-2 antisense MG63 and MC3T3-E1 cell lines. For downregulation of TSPs expression, we prepared antisense constructs for TSP-1 and TSP-2 using the pREP4 an episomal mammalian expression vector, which be able to produce the specific antisense oligonucleotides around chromosome. MG63 and MC3T3-E1 osteoblast-like cells were transfected with the antisense constructs and nonliposomal Fugene 6, and then selected under hygromycin B (50 ${\mu}g/ml$) treatment for 2 weeks. Western blot analysis revealed that expression of the TSP proteins was downregulated in the antisense cell lines. The cell adhesion assay showed that adhesive properties of TSP-1 and TSP-2 antisense MG63 cells on the polystyrene culture plate were reduced to 17% and 21% of the control cells, respectively, and those of the TSP-1 and TSP-2 antisense MC3T3-E1 cells also decreased to 19% and 27% of control, respectively. Adhesion of TSP-1 and TSP-2 antisense MC3T3-E1 cells on Type I collagen-coated culture plate decreased to 27% and 76%, respectively. These results indicate that TSP-1 and TSP-2 proteins may have an important role in adhesion of osteoblast-like cells to extracellular matrix.

Microgrooves on titanium surface affect peri-implant cell adhesion and soft tissue sealing; an in vitro and in vivo study

  • Lee, Hyo-Jung;Lee, Jaden;Lee, Jung-Tae;Hong, Ji-Soo;Lim, Bum-Soon;Park, Hee-Jung;Kim, Young-Kwang;Kim, Tae-Il
    • Journal of Periodontal and Implant Science
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    • v.45 no.3
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    • pp.120-126
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    • 2015
  • Purpose: With the significance of stable adhesion of alveolar bone and peri-implant soft tissue on the surface of titanium for successful dental implantation procedure, the purpose of this study was to apply microgrooves on the titanium surface and investigate their effects on peri-implant cells and tissues. Methods: Three types of commercially pure titanium discs were prepared; machined-surface discs (A), sandblasted, large-grit, acid-etched (SLA)-treated discs (B), SLA and microgroove-formed discs (C). After surface topography of the discs was examined by confocal laser scanning electron microscopy, water contact angle and surface energy were measured. Human gingival fibroblasts (hGFs) and murine osteoblastic cells (MC3T3-E1) were seeded onto the titanium discs for immunofluorescence assay of adhesion proteins. Commercially pure titanium implants with microgrooves on the coronal microthreads design were inserted into the edentulous mandible of beagle dogs. After 2 weeks and 6 weeks of implant insertion, the animal subjects were euthanized to confirm peri-implant tissue healing pattern in histologic specimens. Results: Group C presented the lowest water contact angle ($62.89{\pm}5.66{\theta}$), highest surface energy ($45{\pm}1.2mN/m$), and highest surface roughness ($Ra=22.351{\pm}2.766{\mu}m$). The expression of adhesion molecules of hGFs and MC3T30E1 cells was prominent in group C. Titanium implants with microgrooves on the coronal portion showed firm adhesion to peri-implant soft tissue. Conclusions: Microgrooves on the titanium surface promoted the adhesion of gingival fibroblasts and osteoblastic cells, as well as favorable peri-implant soft tissue sealing.