• Title/Summary/Keyword: Cell Detachment

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A Study on Changes of Cell Adhesive force and Distribution of F-actin and Vinculin under Various Intermittent Hydrostatic Pressure. (간헐적 정수압의 다양한 패턴에 따른 세포 부착력과 F-actin 및 vinculin 의 분포 변화 연구)

  • 김영직;박수아;신호준;김인애;이용재;허수진;황영미;신정욱
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2004.10a
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    • pp.1215-1220
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    • 2004
  • Cell adhesion to any material surface is considered to be fundamental and important phenomenon in the fields of tissue engineering. Cell adhesion molecules, mechanism, and attachment force have been studied and described a lot. However, the effects of mechanical stimuli on the adhesive forces still have been left much to be investigated. In this study, to investigate the changes in cell adhesive force due to resting time period during the intermittent hydrostatic pressurizing (IHP), cells were cultured under the IHP with various resting times. Then the cell adhesive forces were measured quantitatively utilizing a cell detachment test system and immunofluorescent staining was performed using fluorescent microscopy. In the results, immediately after mechanical stimuli (150 minutes after seeding) and one hour later (210 minutes after seeding), the average adhesive force of experimental group 5 (resting time: 15min) compared with that of control group at same culture time was increased significantly (p<0.05). The results indicated that IHP can contribute in improving cell adhesive force and some of time intervals were required for the expression of cell response.

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Gefitinib induces anoikis in cervical cancer cells

  • Byung Chul Jung;Sung-Hun Woo;Sung Hoon Kim;Yoon Suk Kim
    • BMB Reports
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    • v.57 no.2
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    • pp.104-109
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    • 2024
  • Gefitinib exerts anticancer effects on various types of cancer, such as lung, ovarian, breast, and colon cancers. However, the therapeutic effects of gefitinib on cervical cancer and the underlying mechanisms remain unclear. Thus, this study aimed to explore whether gefitinib can be used to treat cervical cancer and elucidate the underlying mechanisms. Results showed that gefitinib induced a caspase-dependent apoptosis of HeLa cells, which consequently became round and detached from the surface of the culture plate. Gefitinib induced the reorganization of actin cytoskeleton and downregulated the expression of p-FAK, integrin β1 and E-cadherin, which are important in cell-extracellular matrix adhesion and cell-cell interaction, respectively. Moreover, gefitinib hindered cell reattachment and spreading and suppressed interactions between detached cells in suspension, leading to poly (ADP-ribose) polymerase cleavage, a hallmark of apoptosis. It also induced detachment-induced apoptosis (anoikis) in C33A cells, another cervical cancer cell line. Taken together, these results suggest that gefitinib triggers anoikis in cervical cancer cells. Our findings may serve as a basis for broadening the range of anticancer drugs used to treat cervical cancer.

Effect of Snake Venom Toxin from Vipera Lebetina Turanica on Neuroblastoma SK-N-SH Cells (Vipera Lebetina Turanica 사독의 신경아세포종 SK-N-SH 세포에 미치는 영향)

  • Sim, Jae-Young;Song, Ho-Sueb
    • Journal of Acupuncture Research
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    • v.25 no.3
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    • pp.53-69
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    • 2008
  • 목적 : 이 연구는 Vipera lebetina turanica의 사독약침액(蛇毒藥鍼液)(Snake venom toxin, SVT)이 인간 신경아세포종의 암세포주인 SK-N-SH 세포에서 암세포성장의 억제 및 그 기전에 대하여 살펴보고자 하였다. 방법 : SVT를 처리한 후 SK-N-SH의 성장억제를 관찰하기 위해 CCK-8 assay와 LDH assay를 시행하였고, apoptosis 평가에는 세포형태의 관찰과 DAPI, TUNEL, Annexin V-PI double staining assay 및 cell detachment assay를 시행하였다. 세포자멸사 관련 세포기전을 보기 위하여 세포주기, 세포내 칼슘량, 세포내 활성산소량 및 미토콘드리아의 세포막전위 변화를 측정하였고, DNA fragmentation assay를 시행하였으며, 세포자멸사 조절 단백인 Bax, Bcl-2, caspase-3, -9의 발현 변화 관찰에는 western blot analysis를 시행하였다. 결과 : SK-N-SH 세포에 SVT를 처리한 후, 신경아세포종 세포의 성장, Apoptosis의 유발 및 기전의 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. SVT를 처리한 SK-N-SH 세포 관찰에서 세포독성은 농도의존적으로 증가하는 경향이 있는 반면 암세포성장의 유의한 억제는 $20{\mu}g/m{\ell}$ SVT에 의해서만 나타났다. 2. 세포자멸사 평가에서 SVT를 처리한 SK-N-SH 세포는 세포자멸사의 특징적 형태를 나타내었다. TUNEL assay에서는 세포자멸사 활성세포가 미약하게 나타난 반면 cell detachment assay와 Annexin V-PI double staining에서는 각각 세포박리와 세포자멸사 활성세포의 유의한 증가를 나타내었다. 3. 세포자멸사 관련 세포기전연구에서 SVT를 처리한 SK-N-SH 세포의 세포주기, 세포 내 칼슘양 및 DNA fragmentation에는 유의한 변화가 관찰되지 않은 반면 세포내 활성산소 양은 유의한 증가를 나타내었고, 그에 따른 미토콘드리아 세포막 전위의 유의한 변동이 관찰되었다. 4. SVT를 처리한 SK-N-SH는 세포자멸사 관련 단백 발현에서 Bax에 대해 유의한 증가를 나타내지 않았으나 caspase-3 및 caspase-9의 유의한 증가와 Bcl-2의 유의한 감소를 나타내었다. 결론 : 이상의 결과는 SVT가 세포 내 활성산소를 증가시킴으로써 미토콘드리아의 세포막전위에 변화를 일으켜 인간 신경아세포종 세포주인 SK-N-SH의 세포박리와 유관한 세포자멸사를 유발함으로써 증식억제 효과가 있음을 입증한 것이다.

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Effect of Snake Venom Toxin from Vipera Lebetina Turanica on Neuroblastoma SK-N-MC Cells (Vipera Lebetina Turanica 사독이 신경아세포종 SK-N-MC 세포에 미치는 영향)

  • Sim, Jae-Young;Song, Ho-Sueb
    • Journal of Acupuncture Research
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    • v.25 no.2
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    • pp.41-57
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    • 2008
  • 목적 : 이 연구는 Vipera lebetina turanica의 사독약침액(蛇毒藥鍼液)(Snake venom toxin, SVT)이 인간 신경아세포종의 암세포주인 SK-N-MC 세포에서 암세포성장의 억제 및 그 기전에 대하여 살펴보고자 하였다. 방법 : SVT를 처리한 후 SK-N-MC의 성장억제를 관찰하기 위해 CCK-8 assay와 LDH assay를 시행하였고, apoptosis 평가에는 세포형태의 관찰과 DAPI, TUNEL, Annexin V-PI double staining assay 및 cell detachment assay를 시행하였다. 세포자멸사 관련 세포기전을 보기 위하여 세포주기, 세포내 칼슘량, 세포내 활성산소량 및 미토콘드리아의 세포막전위 변화를 측정하였고, DNA fragmentation assay를 시행하였으며, 세포자멸사 조절 단백인 Bax, Bcl-2, caspase-3, -9의 발현 변화 관찰에는 western blot analysis를 시행하였다. 결과 : SK-N-MC 세포에 SVT를 처리한 후, 신경아세포종 세포의 성장, Apoptosis의 유발 및 기전에 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. SVT를 처리한 SK-N-MC 세포 관찰에서 $20{\mu}g/m{\ell}$ SVT 처리가 암세포성장의 유의한 억제를 나타내었다. 세포독성 관찰에서 SVT처리는 처리하지 않은 것에 비하여 증가를 나타내었다. 2. 세포자멸사 평가에서 SVT를 처리한 SK-N-MC 세포는 세포자멸사의 특징적 형태를 나타내었다. TUNEL assay에서는 세포자멸사 활성세포가 미약하게 나타난 반면 cell detachment assay와 Annexin V-PI double staining에서는 각각 세포박리와 세포자멸사 활성세포의 유의한 증가를 나타내었다. 3. 세포자멸사 관련 세포기전연구에서 SVT를 처리한 SK-N-MC 세포의 세포주기, 세포내 칼슘량 및 DNA fragmentation에는 유의한 변화가 관찰되지 않은 반면 세포내 활성산소 양은 유의한 증가를 나타내었고, 그에 따른 미토콘드리아 세포막 전위의 유의한 변동이 관찰되었다. 4. SVT를 처리한 SK-N-MC는 세포자멸사 관련 단백 발현에서 caspase-9에 대해 유의한 증가를 나타내지 않았으나 Bax 및 caspase-3의 유의한 증가와 Bcl-2의 유의한 감소를 나타내었다. 결론 : 이상의 결과는 SVT가 세포내 활성산소를 증가시키므로 미토콘드리아의 세포막전위에 변화를 일으켜 인간 신경아세포종 세포주인 SK-N-MC의 세포박리와 유관한 세포자멸사를 유발하므로 증식억제 효과가 있음을 입증한 것이다.

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Effects of Epigallocatechin Gallate on Adhesion, Invasion and Matrix Metalloproteinase Activity in MDA-MB-231 Human Breast Cancer Cells (Epigallocatechin Gallate(EGCG)가 MDA-MB-231 인체 유방암 세포의 부착성, 침윤성과 Matrix Metalloproteinase 활성에 미치는 영향)

  • Bang Myung Hee;Kim Ji Hye;Kim Woo Kyoung
    • Journal of Nutrition and Health
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    • v.38 no.2
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    • pp.104-111
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    • 2005
  • Tumor invasion is composed of four steps: cell adhesion to the extracellular matrix, degradation of the extracellular matrix components, tumor cell motility followed by cell detachment. Matrix metalloproteinases (MMPs) are important proteinases that associated with degradation of matrix component. Epigallocatechin gallate (EGCG) is a major polyphenotic constituent of green tea. In the study, we examined the anti-invasive and MMP activity suppression effects of EGCG in MDA-MB-231 human breast cancer cells. MDA-MB-23l human breast cancer cells were cultured with various concentrations 0 - 100 μM of EGCG. EGCG significantly inhibited the cell adhesion to the fibronectin. Cell motility through gelatin filter and invasion to Matrigel were inhibited dose-dependently by EGCG treatment. EGCG also inhibited the activities of MMP-2, -9 and the amount of MMP-9 (α = 0.05). Therefore, EGCG may contribute to the potential beneficial food component to prevent the invasion and metastasis in breast cancer. (Korean J Nutrition 38(2): 104~111, 2005)

Non-classical role of Galectin-3 in cancer progression: translocation to nucleus by carbohydrate-recognition independent manner

  • Kim, Seok-Jun;Chun, Kyung-Hee
    • BMB Reports
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    • v.53 no.4
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    • pp.173-180
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    • 2020
  • Galectin-3 is a carbohydrate-binding protein and regulates diverse functions, including cell proliferation and differentiation, mRNA splicing, apoptosis induction, immune surveillance and inflammation, cell adhesion, angiogenesis, and cancer-cell metastasis. Galectin-3 is also recommended as a diagnostic or prognostic biomarker of various diseases, including heart disease, kidney disease, and cancer. Galectin-3 exists as a cytosol, is secreted in extracellular spaces on cells, and is also detected in nuclei. It has been found that galectin-3 has different functions in cellular localization: (i) Extracellular galectin-3 mediates cell attachment and detachment. (ii) cytosolic galectin-3 regulates cell survival by blocking the intrinsic apoptotic pathway, and (iii) nuclear galectin-3 supports the ability of the transcriptional factor for target gene expression. In this review, we focused on the role of galectin-3 on translocation from cytosol to nucleus, because it happens in a way independent of carbohydrate recognition and accelerates cancer progression. We also suggested here that intracellular galecin-3 could be a potent therapeutic target in cancer therapy.

Electron and Light Microscopic Studies on the Development of Oidia from Somatic Mycelium of Coprinus cinereus

  • Yoon, Kwon-S.
    • Mycobiology
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    • v.32 no.4
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    • pp.164-169
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    • 2004
  • Development of oidia, a type of thallic spores from monokaryotic mycelium of Coprinus cinereus was examined with electron microscope and light microscopes. Oidia formation in this fungus is unique in its mode of formation compared with other types of asexual sporogenesis. Oidiogenesis in C. cinereus is carried out in three steps: 1) Formation of oidiophore from the parent mycelium, 2) Formation of initials of oidial cells from swollen oidiophore, 3) Segmentation and detachment of mature oidial cell. Oidiophores appear to spring out singly as a swollen hyphal branches from the normal foot hyphae or sometimes coiled hypha. From the oidiophore, oidial branches sprout out forming a group of $2{\sim}6$, most often 4 oidial cells and each oidial cell undergoes a single mitosis resulting in 2 oidia. One of the sibling oidial cells in a group is frequently transformed into a new oidiophore, thus oidiogenic structures are tandemly produced at the several different levels.

Spatio-temparal Pattern Formation of Abdominal Muscle in Xenopus Iaevis

  • Ko, Che-Myong;Chung, Hae-Moon
    • Animal cells and systems
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    • v.1 no.2
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    • pp.329-335
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    • 1997
  • The final pattern of the skeletal muscle of a vertebrate depends on the position-specific behavior of the muscle precursor cells during early developmental process and the abdominal muscle is made of cells which migrate a relatively long distance from their original tissue, myotome of dorsal mesoderm. We report the spatia-temporal migration pattern of abdominal muscle in Xenopus laevis by in situ hybridization and immunohistological studies. Shortly after hatching tadpole stage (stage 31/32), a group of myotomal cells detaches from the lower tip of the second somite and migrates ventrally to the lower position of abdomen. At stage 34/35, a second cell group migrates away from the third somite. Total 7 myotomal cell groups migrate ventrally one by one from the second to eighth myotome along their own pathways through the cell free space located between epidermis and subepidermal layer of the abdomen. During migration, the sizes of the cell groups (abdominal muscle anlagens) are increased to several tens fold. Around stage 40 all the abdominal muscle anlagens reaches their final positions and are interconnected side by side rostrocaudally. They are also connected to other types of muscles, forming a large multisegmented abdominal muscle. Heat shock study suggests that the disruption of segmentation of somites does not block the detachment of abdominal muscle anlagen, though the treatment gave stage- and dosagedependent effects on the migration speed.

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Bead-to-Bead Cell Transfer by Induction of Detachment of Anchorage Dependent HeLa Cells Grown on Macroporous Microcarriers (부착성 HeLa 세포의 탈리 유도에 의한 다공성 미립담체의 담체간 전이 배양)

  • 이두훈;박정극
    • KSBB Journal
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    • v.13 no.1
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    • pp.83-89
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    • 1998
  • Using a cellulose macroporous microcarrier, HeLa cells were cultivated in 100mL spinner flask(Bellco Co., USA) and confluent cell laden microcarriers were subcultured by bead-to-bead cell transfer method. In macroporous mcirocarrier-HeLa system viable suspended cells played an important role in bead-to-bead cell transfer and that could be increased by use of RPMI-1640, a calcium-ion-reduced-media and high speed agitation. Successful bead-to-bead cell transfers were performed continuously three time in spinner flask. We applied this technique to produce recombinant Vaccinia virus which express $\beta$-galactosidase. Recombinant protein yield of bead-to-bead transferred culture was comparable to conventional microcarrier cultures that were inoculated by cells detached from T-flask. Although trypsinization is a useful method for subculturing microcarriers in some cases, that process adds quality control problem and handling steps to large scale cell production. There fore, bead-to-bead cell transfer technique offers another convenient and efficient scale-up method for continuous microcarrier cultures.

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