• 한국동물학회지
• /
• 제38권4호
• /
• pp.542-549
• /
• 1995

Extracellular matrix(ECM) 단백질이 SK-N-SH 및 IMR-32 세포주가 신경계 세포로 분화되는 데 미치는 영향을 조사하였다. Laminin과 collagen으로 도말한 배양기에서 7일간 배양했을 때 SK-N-SH세포는 잘 발달된 신경측색생장을 보였으나 IMR-32세포는 뚜렷한 형태변화를 나타내지 않았다. 왜 IMR-32세포가 ECM 단백질에 반응을 하지 않는가를 규명하기 위하여 ECM단백질에 의한 초기 신호전달기작을 두 세포주에서 분석하였다. ECM 단백질을 도말한 배양기에 세포를 깔았을 때 한시간 만에 tyrosine 인산화된 단백질이 두 세포 모두 증가함을 볼 수 있었다. 아울러 focal adhesion kinase(FAK)의 tyrosine 인산화도 두 세포주 모두에서 증가하였다. 이러한 결과는 두 세포주가 ECM 단백질에 의한 초기 신호전달체계가 정상임을 의미한다. 신경세포 분화과정에 증가한다고 알려진 Bcl-2 및 NSE의 량을 ECM 단백질 처리후 조사하였을 때 SK-N-SH 세포주는 두 단백질이 증가 했지만 IMR-32 세포주는 변화가 없었다. 이러한 결과는 IMR-32 세포주가 ECM 단백질에 반응하지 않는 것이 ECM 단백질에 의한 신호전달체계에 문제가 있다기 보다 신경계세포로 분화되는 데 필요한 유전인자의 발현조절에 문제가 있음을 시사한다.

• 대한기계학회:학술대회논문집
• /
• 대한기계학회 2008년도 추계학술대회A
• /
• pp.1455-1457
• /
• 2008

Cell adhesion is a coordinated process involving initial binding of integrin receptors to extracellular matrix (ECM), recruitment of adhesion proteins, and focal adhesion assembly. The formation of mechanically stable focal adhesion assembly of cells within surrounding ECM is a key parameter to direct numerous cellular functions including cell migration, differentiation, and apotosis. With current cell adhesion assays, it is difficult to understand contributions of each coordinated event on evolution of cell adhesion strengthening since cells spontaneously spread upon their adhesion to the substrate, thus remodeling their cytoskeletal structure. In this presentation, novel approaches for analysis of cell adhesion strengthening process based on the combination of mechanical device, micro-patterned substrates, and molecular biological techniques will be discussed.

• Journal of Microbiology and Biotechnology
• /
• 제9권1호
• /
• pp.119-121
• /
• 1999

Tumor cell interaction with the extracellular matrix is defined as the critical event of tumor invasion that signals the initiation of a metastatic cascade. Sesquicillin has been identified as an inhibitor of melanoma cell adhesion to the components of the extracellular matrix (ECM) in cultured broth of fungal strain F60063. Sesquicillin strongly inhibited the adhesion of B16 melanoma cells to laminin, fibronectin, and typeIV collagen. It also inhibited B16 melanoma cell invasion of reconstituted basement membrane Matrigel in vitro in a dose-dependent manner. These results suggest that sesquicillin is a new class of nonpeptidic ECM adhesion inhibitor having anti-invasive activity.

• Journal of Microbiology and Biotechnology
• /
• 제11권6호
• /
• pp.922-927
• /
• 2001

In an effort to regulate the mammalian cell behavior in entrapment with a gel, we have functionalized hydrogels with the putative cell-binding (-Arg-Gly-Asp-)(RGD) domain. An adhesion molecule of Gly-Arg-Gly-Asp-Ser (GRGDS) peptides, a cell recognition ligand, was induced into thermo-reversible hydrogels, composed of N-isopropylacrylamide with small amounts of acrylic acid (typically 2-5 $mol\%$ in feed), as a biomimetic extracellular matrix (ECM). The GRGDS containing a p(NiPAAm-co-AAc) copolymer gel was studied in vitro for its ability to promote the spreading and viability of cells by introducing a GRGDS sequence. Hydrogel with no adhesion molecule was a poor ECM for adhesion, permiting spreading of only $3\%$ of the seeded cells for 36h. By immobilizing the peptide linkage into the hydrogel, the conjugation of RGD promoted $50\%$ of proliferation for 36h. However, the GREDS sequence, nonadhesive peptide linkage, conjugated hydrogel showed only $5\%$ of the seeded cell for the same time period. In addition, with the serum-free medium, only GRGDS peptides conjugated to hydrogel was able to promotecell spreading, while there was no cell proliferation in the hydrogel without GRGDS. Thus, the GRGDS peptide-conjugated thermo-reversible hydrogel specifically mediated the cell spreading. This result suggests that utilization of peptide sequences conjugating with the cell-adhesive motifs can enhance the degree of cell surface interaction and influence the long-term formation of ECM in vitro.

• BMB Reports
• /
• 제45권3호
• /
• pp.159-164
• /
• 2012

CD99 is known to be involved in the regulation of cell-cell adhesion. However, it remains unclear whether CD99 controls cell-extracellular matrix adhesion. In this study, the effects of CD99 activation on cell-extracellular matrix adhesion were investigated. It was found that engagement of CD99 with the stimulating antibody YG32 downregulated the adhesion of MCF-7 cells to fibronectin, laminin and collagen IV in a dose-dependent manner. The CD99 effect on cell-ECM adhesion was inhibited by overexpression of the dominant negative form of CD99 or CD99 siRNA transfection. Treatment of cells with $Mn^{2+}$ or by ${\beta}_1$ integrin-stimulating antibody restored the inhibitory effect of CD99 on cell-ECM adhesion. Cross-linking CD99 inactivated ${\beta}_1$ integrin through conformational change. CD99 activation caused dephosphorylation at Tyr-397 in FAK, which was restored by the ${\beta}_1$ stimulating antibody. Taken together, these results provide the first evidence that CD99 inhibits cell-extracellular matrix adhesion by suppressing ${\beta}_1$ integrin affinity.

• 생약학회지
• /
• 제31권4호
• /
• pp.394-400
• /
• 2000

Tumor cell interaction with the extracellular matrix (ECM) is defined as the critical event of tumor invasion that signals the initiation of a metastatic cascade. To search for anti-metastatic agent from plants, several plant extracts were screened by cell- ECM anti-adhesion test. As result, Boehmeria pannosa, Dryopteris crassirhizoma, Scilla scilloides, and Agrimonia pilosa were shown a significant anti-adhesion activity.

• 생명과학회지
• /
• 제32권2호
• /
• pp.148-154
• /
• 2022

국소접착인산화효소(FAK)는 국소접착부에서 세포부착, 세포이동, 세포역학적 신호전달 등에 관여한다고 알려져 있다. 그러나 세포 외 기질(ECM)과 상호작용하는 인테그린 막단백질과 함께 위치하는 세포막 미세영역(membrane microdomain)의 종류와 ECM 구성에 따른 FAK 활성은 여전히 불분명하다. 형광 공명 에너지 전달(FRET)을 기반으로 유전적으로 인코딩 된 바이오센서는 세포 내 FAK 신호를 높은 시공간 해상도로 제공할 수 있다. 본 연구에서는 유리, 제1형 콜라겐, 피브로넥틴, 라미닌의 ECM 조건에서 FRET 기반 막 표적 FAK 바이오센서를 사용하여 지질유동섬(Lipid raft) 및 비-지질유동섬(non-Lipid raft)에서 FAK의 활성을 분석하고 시각화 하였다. 흥미롭게도, 지질유동섬에서 라미닌 조건 하의 FAK 활성은 다른 ECM 조건보다 낮았고, 비-지질유동섬에서 FAK 활성은 다른 ECM 조건보다 낮았다. 동일한 ECM 조건 상의 비교에서는 피브로넥틴 조건일 때 지질유동섬에서 비-지질유동섬 보다 높은 FAK 활성이 관측되었다. 따라서 이번 연구는 FAK 활성도가 ECM 유형 및 세포막 미세영역에 따라 특이적으로 조절되는 것을 시각적, 정량적으로 보여준다.

• 한국진공학회:학술대회논문집
• /
• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
• /
• pp.275-275
• /
• 2013

Osteoblast is one of cells related with osseointegration and many research have conducted the adhesion of osteoblast onto the surface of implant. In the osseointegration, biocompatibility of the implant and cell adhesion to the surface are important factors. The researches related to cell adhesion have a direction from micro-scaled surface roughness to nano-scaled surface roughness with advancing nanotechnology. A cell reacts and sense to stimuli from extracellular matrix (ECM) and topography of the ECM [1]. Thus, for better osseointegration, we should provide an environment similar to ECM. In this study, we synthesize TiO2 nanowires using hydrothermal reaction because TiO2 provides inertness to titanium on its surface and enables it used as an implant material for the orthopedic treatment such as fixation of the bone fracture [2]. Ti substrate is immersed into NaOH aqueous solution. The solution are heated at $140{\sim}200^{\circ}C$ for various time (10~720 minutes). After heat treatment, we take out the sample and immerse it into HCl aqueous solution for 1 hour. The acid treated sample is heated again at $500^{\circ}C$ for 3 hours [3]. Then, we culture osteoblast on the TiO2 nanowires. For investigating cell adhesion onto nanostructured surface, we conduct several tests such as MTT assay, ALP (Alkaline phosphatase) activity assay, measuring calcium expression, and so on. These preliminary results of the cell culture on the nanowires are foundation for investigating cell-material interaction especially with nanostructure interaction.

• 한국동물생명공학회지
• /
• 제38권3호
• /
• pp.121-130
• /
• 2023

Background: Coating a culture plate with molecules that aid in cell adhesion is a technique widely used to produce animal cell cultures. Extracellular matrix (ECM) is known for its efficiency in promoting adhesion, survival, and proliferation of adherent cells. Gelatin, a cost-effective type of ECM, is widely used in animal cell cultures including feeder-free embryonic stem (ES) cells. However, the optimal concentration of gelatin is a point of debate among researchers, with no studies having established the optimal gelatin concentration. Methods: In this study, we coated plastic plates with gelatin in a concentration-dependent manner and assessed Young's modulus using atomic force microscopy (AFM) to investigate the microstructure of the surface of each plastic plate. The adhesion, proliferation, and differentiation of the ESCs were compared and analyzed revealing differences in surface microstructure dependent on coating concentration. Results: According to AFM analysis, there was a clear difference in the microstructure of the surface according to the presence or absence of the gelatin coating, and it was confirmed that there was no difference at a concentration of 0.5% or more. ES cell also confirmed the difference in cell adhesion, proliferation, and differentiation according to the presence or absence of gelatin coating, and also it showed no difference over the concentration of 0.5%. Conclusions: The optimum gelatin-coating for the maintenance and differentiation of ES cells is 0.5%, and the gelatin concentration-mediated microenvironment and ES cell signaling are closely correlated.

• International Journal of Oral Biology
• /
• 제32권3호
• /
• pp.93-101
• /
• 2007

Cell behavior of the transformed cells is known to affect by interaction with extracellular matrix (ECM) proteins and integrin. To investigate the alterations of both integrin expression and cell-matrix interaction during neoplastic conversion of human oral kerationcytes, we studied expression levels of integrin subunits by flow cytometry and cellular responses to the ECM proteins in normal human oral keratinocytes (NHOKs), HPV-immortalized HOK-16B line, and three oral cancer cell lines established from HOK-16B line, CTHOK-16B-BaP, CTHOK-16B-DMBA, and CTHOK-16B-Dexa lines. The expression levels of ${\alpha}\;and\;{\beta}$ integrin subunits were shown decreased tendency in human oral keratinocytes undergoing immortalization and tumorigenic transformation except CTHOK-16B-DMBA line tested. Although ${\alpha}v{\beta}6$ integrin is known to be highly expressed in squamous cell carcinomas, and the altered integrin expression is suspected to be associated with cellular carcinogenesis, ${\alpha}v$ integrin subunit and ${\alpha}v{\beta}6$ integrin did not express in oral cancer cell lines tested. Cell behavior to the ECM proteins in HOK-16B line was generally similar to that of exponentially proliferating NHOKs. The adhesion activity profiles of type I collagen were very similar to that of its laminin counterparts, but fibronectin showed minimal adhesion activity under our conditions compared to the BSA control. The ability of the CTHOK-16B-BaP line to spread upon type I collagen and laminin markedly decreased, but migration was notably increased on type I collagen. In contrast, CTHOK-16B-DMBA and CTHOK-16B-Dexa lines spread less but migrated more upon type I collagen than immortalized HOK-16B line. These data indicate that downregulation of integrin subunits causes the changes of cellular responses to the ECM proteins during neoplastic conversion of human oral keratinocytes, and that cellular responses to the ECM proteins in oral cancer cell lines established by exposing different carcinogens are variable according to chemical carcinogens treatment.