• 제목/요약/키워드: Cell Migration Assay

검색결과 283건 처리시간 0.03초

소망막내피세포에서 금 나노입자의 최종당화산물에 의한 세포 이동 및 침윤성 억제 효과 (Gold Nanoparticles Inhibit AGEs Induced Migration and Invasion in Bovine Retinal Endothelial Cells)

  • 채수철
    • 환경생물
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    • 제28권1호
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    • pp.8-13
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    • 2010
  • 본 연구는 BRECs에서 AGEs로 유도된 세포의 이동 및 침윤에 있어서 AuNP의 역할에 관한 연구이다. 소 망막으로부터 내피세포를 분리하고, 세포 생존율은 MTT assay로 확인하였다. Wound migration assay는 BRECs의 이동력을 확인하기 위해 수행하였다. Tube formation은 on-gel system을 통해 확인하였다. AuNP의 apoptosis 유도는 caspase-3 assay를 통해 확인하였다. AGE-BSA은 세포증식 및 이동에 있어서 증가함을 보여주었다. 또한 AuNP는 AGE-BSA 존재 유무에 상관없이 세포의 증식, 이동, 신생혈관 형성을 억제하였고, caspase-3을 통해 apoptosis를 유도하였다. 이러한 결과, AuNP는 AGE로 유도된 신생혈관 형성 및 세포의 이동성을 억제하는 약재제로서, 당뇨성 합병증에 있어서 잠재적으로 중요한 분자가 될 것이다.

약물 담지 다공성 중공 실리카 미세구 주위 세포의 주화성 이동 (Chemotactic Cell Migration around Hollow Silica Beads Containing Chemotatic Reagent)

  • 김해춘;강미선;이석우
    • KSBB Journal
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    • 제25권4호
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    • pp.344-350
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    • 2010
  • This paper demonstrates a microfluidic chip incorporating patterned hollow silica beads that can be effectively used for chemotaxis assay. The hollow silica bead has been exploited to develop a carrier for chemoattractant to induce cell migration. The microfluidic chip contains a patterned array of microfabricated docks which can hold only one bead per docking site. The hollow bead placed inside microfluidic chip releases chemotactic reagent (PDGF-BB) around its periphery in a controlled fashion which generates a signal for chemotatic migration of fibroblast cells. The number of cells migrated close to each bead has been assessed. On-chip cell migration assay showed a remarkable result proving the high efficiency and reliable accuracy in quantitative analysis. Therefore, the device could be extensively used in cell migration assay and other various studies related to cellular movements.

C6 Glioma 세포에서 Protein Kinase C Alpha 발현 저해를 통한 송절 약침액의 이주 억제 효과 (Pinus densiflora Gnarl Inhibits Migration through Suppression of Protein Kinase C in C6 Glioma Cells)

  • 민일국;이강파;장해룡;문진영
    • Korean Journal of Acupuncture
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    • 제32권2호
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    • pp.51-58
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    • 2015
  • Objectives : Pinus densiflora gnarl, called Song-Jeol in Korean medicine, has been used to cure inflammatory diseases such as arthritis. In the present study, we evaluated inhibitory property of Song-Jeol pharmacopuncture(SJ) on C6 glioma cell migration. Methods : To evaluate cell viability on C6 glioma cells of SJ, the viability was assessed by using Ez-cytox assay kit. The cell migration was assessed by wound-healing assay and Boyden chamber assay, respectively. LPS-induced NO productions were determined by using the Griess reagent. The expression of iNOS and protein kinase $C(PKC)-{\alpha}$ were estimated by western blotting assay. Results : In the wound-healing assay and Boyden chamber assay, SJ showed a significant inhibition on serum-induced C6 glioma cell migration. In addition, NO production was decreased by SJ through suppression of iNOS expression in LPS-stimulated C6 glioma cell. Futhermore, LPS-induced protein kinase $C(PKC)-{\alpha}$ expression was effectively inhibited by SJ. Conclusions : These results demonstrated that SJ was useful for the suppression of the C6 glioma cell migration.

Auraptene Inhibits Migration and Invasion of Cervical and Ovarian Cancer Cells by Repression of Matrix Metalloproteinasas 2 and 9 Activity

  • Jamialahmadi, Khadijeh;Salari, Sofia;Alamolhodaei, Nafiseh Sadat;Avan, Amir;Gholami, Leila;Karimi, Gholamreza
    • 대한약침학회지
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    • 제21권3호
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    • pp.177-184
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    • 2018
  • Objectives: Auraptene, a natural citrus coumarin, found in plants of Rutaceae and Apiaceae families. In this study, we investigated the effects of auraptene on tumor migration, invasion and matrix metalloproteinase (MMP)-2 and -9 enzymes activity. Methods: The effects of auraptene on the viability of A2780 and Hela cell lines was evaluated by MTT assay. Wound healing migration assay and Boyden chamber assay were determined the effect of auraptene on migration and cell invasion, respectively. MMP-2 and MMP-9 activities were analyzed by gelatin zymography assay. Results: Auraptene reduced A2780 cell viability. The results showed that auraptene inhibited in vitro migration and invasion of both cells. Furthermore, cell invasion ability suppressed at $100{\mu}M$ auraptene in Hela cells and at 25, $50{\mu}M$ in A2780 cell line. Gelatin zymography showed that for Hela cell line, auraptene suppressed MMP-2 enzymatic activity in all concentrations and for MMP-9 at a concentration between 12.5 to $100{\mu}M$ in A2780 cell line. Conclusion: Auraptene inhibited migration and invasion of human cervical and ovarian cancer cells in vitro by possibly inhibitory effects on MMP-2 and MMP-9 activity.

DNA damage와 Apoptosis를 정량화하는 단세포전기영동법 (Single Cell Gel Electrophoresis (comet assay) to Detect DNA Damage and Apoptosis in Cell Level)

  • 류재천;김현주;서영록;김경란
    • 한국환경성돌연변이발암원학회지
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    • 제17권2호
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    • pp.71-77
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    • 1997
  • The single cell gel electrophoressis(SCGE) assay, also known as the comet assay, is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakage in mammalian cells. The SCGE or comet assay is a promising test for the detection of DNA damage and repair in individnal cells. It has widespread potential applications in DNA damage and repair studies, genotoxicity testing and biomonitoring. In this microgel electrophoresis technique, cells are embedded in agarose gel on microscope slides, iysed and electrophoresed under alkaline conditions. Cells with increased DNA damage display increased migration of DNA from the nucleus towards the anode. The length of DNA migration indicates the amount of DNA breakage in the cell. The comet assay is also capable of identifying apoptotic cells which contain highly fragmented DNA. Here we review the development of the SCGE assay, existing protocols for the detection and analysis of comets, the relevant underlying principles determining the behaviour of DNA and the potential applications of the technique.

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대계 약침액의 C6 신경교종 세포에 대한 이주 억제 효과 (The Anti-Migratory Effect of Cirsium japonicum Pharmacopuncture in C6 Glioma Cell)

  • 박주연;이강파;장해룡;문진영
    • Korean Journal of Acupuncture
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    • 제30권4호
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    • pp.212-219
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    • 2013
  • Objectives : Cirsium japonicum is a traditional Korean medicine that has been used in the treatment of inflammatory diseases such as appendicitis, hepatitis, pulmonary abscess and tumor. The aim of study was to elucidate anti-migratory activity of CJP(Cirsium japonicum pharmacopuncture) through regulation of inflammatory mediators in C6 glioma cell. Methods : Nitric oxide(NO) production was determined by using nitrite assay. The cell migration was analyzed by wound-healing assay and Boyden chamber assay. The expression levels of iNOS, and protein kinase C(PKC)-${\alpha}$ were measured by western blotting assay. Results : CJP showed a significant decrease on NO production. Moreover, glioma cell migration was effectively suppressed by CJP. Furthermore, CJP inhibited the expressions of iNOS and PKC-${\alpha}$ in C6 glioma cells. Conclusions : These results suggest that CJP inhibits glioma cell migration and iNOS expression through regulation of PKC-${\alpha}$. Therefore, it is expected that CJP could be an effective agents for blocking malignant progression of glioma.

Primary Cilium by Polyinosinic:Polycytidylic Acid Regulates the Regenerative Migration of Beas-2B Bronchial Epithelial Cells

  • Gweon, Bomi;Jang, Tae-Kyu;Thuy, Pham Xuan;Moon, Eun-Yi
    • Biomolecules & Therapeutics
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    • 제30권2호
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    • pp.170-178
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    • 2022
  • The airway epithelium is equipped with the ability to resist respiratory disease development and airway damage, including the migration of airway epithelial cells and the activation of TLR3, which recognizes double-stranded (ds) RNA. Primary cilia on airway epithelial cells are involved in the cell cycle and cell differentiation and repair. In this study, we used Beas-2B human bronchial epithelial cells to investigate the effects of the TLR3 agonist polyinosinic:polycytidylic acid [Poly(I:C)] on airway cell migration and primary cilia (PC) formation. PC formation increased in cells incubated under serum deprivation. Migration was faster in Beas-2B cells pretreated with Poly(I:C) than in control cells, as judged by a wound healing assay, single-cell path tracking, and a Transwell migration assay. No changes in cell migration were observed when the cells were incubated in conditioned medium from Poly(I:C)-treated cells. PC formation was enhanced by Poly(I:C) treatment, but was reduced when the cells were exposed to the ciliogenesis inhibitor ciliobrevin A (CilioA). The inhibition of Beas-2B cell migration by CilioA was also assessed and a slight decrease in ciliogenesis was detected in SARS-CoV-2 spike protein (SP)-treated Beas-2B cells overexpressing ACE2 compared to control cells. Cell migration was decreased by SP but restored by Poly(I:C) treatment. Taken together, our results demonstrate that impaired migration by SP-treated cells can be attenuated by Poly(I:C) treatment, thus increasing airway cell migration through the regulation of ciliogenesis.

Compound K attenuates stromal cell-derived growth factor 1 (SDF-1)-induced migration of C6 glioma cells

  • Kim, Hyuck;Roh, Hyo Sun;Kim, Jai Eun;Park, Sun Dong;Park, Won Hwan;Moon, Jin-Young
    • Nutrition Research and Practice
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    • 제10권3호
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    • pp.259-264
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    • 2016
  • BACKGROUND/OBJECTIVES: Stromal cell-derived growth factor 1 (SDF-1), also known as chemokine ligand 12, and chemokine receptor type 4 are involved in cancer cell migration. Compound K (CK), a metabolite of protopanaxadiol-type ginsenoside by gut microbiota, is reported to have therapeutic potential in cancer therapy. However, the inhibitory effect of CK on SDF-1 pathway-induced migration of glioma has not yet been established. MATERIALS/METHODS: Cytotoxicity of CK in C6 glioma cells was determined using an EZ-Cytox cell viability assay kit. Cell migration was tested using the wound healing and Boyden chamber assay. Phosphorylation levels of protein kinase C $(PKC){\alpha}$ and extracellular signal-regulated kinase (ERK) were measured by western blot assay, and matrix metallopeptidases (MMP) were measured by gelatin-zymography analysis. RESULTS: CK significantly reduced the phosphorylation of $PKC{\alpha}$ and ERK1/2, expression of MMP9 and MMP2, and inhibited the migration of C6 glioma cells under SDF-1-stimulated conditions. CONCLUSIONS: CK is a cell migration inhibitor that inhibits C6 glioma cell migration by regulating its downstream signaling molecules including $PKC{\alpha}$, ERK1/2, and MMPs.

Influence of Curcumin on HOTAIR-Mediated Migration of Human Renal Cell Carcinoma Cells

  • Pei, Chang-Song;Wu, Hong-Yan;Fan, Fan-Tian;Wu, Yi;Shen, Cun-Si;Pan, Li-Qun
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권10호
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    • pp.4239-4243
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    • 2014
  • Background: This study investigated the influence of curcumin on HOX transcript antisense RNA (HOTAIR)-mediated migration of cultured renal cell carcinoma (RCC) cells. Materials and Methods: Five RCC cell lines (769-P, 769-P-vector, 769-P-HOTAIR, 786-0, and Kert-3 ) were maintained in vitro. The expression of HOTAIR mRNA was determined by quantitative real-time PCR and cell migration was measured by transwell migration assay. The effects of different concentrations of curcumin (0 to $80{\mu}mol/L$) on cell proliferation was determined by the CCK-8 assay and influence of non-toxic levels (0 to $10{\mu}M$) on the migration of RCC cells was also determined. Results: Comparison of the 5 cell lines indicated a correlation between HOTAIR mRNA expression and cell migration. In particular, the migration of 769-P-HOTAIR cells was significantly higher than that of 769-P-vector cells. Curcumin at $2.5-10{\mu}M$ had no evident toxicity against RCC cells, but inhibited cell migration in a concentration-dependent manner. Conclusions: HOTAIR expression is correlated with the migration of RCC cells, and HOTAIR may be involved in the curcumin-induced inhibition of RCC metastasis.

곡기생(槲寄生)의 항염증 효능 및 암세포 이주저해에 미치는 영향 (The Effects of Gokgisaeng on Anti-inflammation and Rat C6 Glioma Cell Migration)

  • 김현영;장수영;정지천;신현철
    • 대한한방내과학회지
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    • 제34권1호
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    • pp.31-45
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    • 2013
  • Objectives : Gokgisaeng (Korean mistletoe) is used for the treatment of inflammatory and cancer diseases in traditional Korean medicine and its major component lectins have been reported to induce nitric oxide (NO) in RAW 264.7 macrophages, and also induce apoptosis of various types of cancer cells, although its modulatory effects on cancer cell migration and macrophage activation is poorly understood. The aim of this study is to clarify molecular mechanisms of action responsible for the anti-inflammatory and antitumor migration potentials of Korean mistletoe extract (KME). Methods : We investigated the anti-inflammatory activity of KME on NO production and inducible nitric oxide synthase (iNOS) expression by lipopolysaccharide (LPS) in both RAW 264.7 macrophages and rat C6 glioma cells, and also evaluated inhibitory efficacy on glioma cell growth and migration. For assessment, XTT assay, nitrite assay, RT-PCR, scratch-wound and Boyden chamber assay, and western blot analysis were performed. Results : Previously reported, unlike the efficacy of Gokgisaeng lectin, KME inhibited NO production and iNOS expression, and suppressed pro-inflammatory mediators including IL-$1{\beta}$, IL-6, COX-2, iNOS in LPS-stimulated RAW 264.7 cells. Furthermore, KME suppressed tumor cell growth and migration, and it also inhibited LPS-induced NO release and iNOS activation by down-regulating expression of protein kinase C (PKC) and phosphorylation of ERK in C6 glioma cells. Conclusions : Our research findings provide evidence that KME can play a significant role in blocking pro-inflammatory reaction and malignant progression of tumors through the suppression of NO/iNOS by down-regulating of inflammatory signaling pathways, PKC/ERK.