대장암 세포주 KM12C 및 아세포주의 흡착력과 당구조

이봉화,김영식 大韓消化器病學會 1998 大韓消化器病學會誌 Vol.32 No.4

CD29 및 CD98 활성 매개에 의한 Jurkat T 세포의 유착과 그 활용

김병훈,조재열,Kim, Byung-Hun,Cho, Jae-Youl 대한약학회 2009 약학회지 Vol.53 No.3

Cell-cell adhesion managed by various adhesion molecules plays an important role in regulating functional activation of cells. This event mediates attachment of inflammatory cells to endothelial cells, interaction of antigen-presenting cells with T cells and metastatic adherence of cancer cells to epithelial tissue cells. Therefore, this cellular response is considered as one of therapeutic target to treat various cancers and inflammatory diseases. To develop proper model for evaluation of functional activation of adhesion molecules, the ability of U937 and Jurkat T cells responsive to various adhesion inducers such as phorbal-12-myristate-13-acetate (PMA), staurosporin and monoclonal antibodies to CD29, CD43 and CD98 was investigated using quantitative cell-cell adhesion assay. U937 cells made more cell-cell clusters by the treatment of antibodies to CD29 and CD43 than Jurkat T cells, while Jurkat T cells exhibited increased cell-cell adhesion ability in CD98 antibody treatment. In agreement, the surface levels of CD29 and CD98 were highly observed in U937 and Jurkat T cells, respectively. Therefore, our data suggest that Jurkat T and U937 cells can be used for model system to evaluate functional activation of adhesion molecules such as CD29 and CD98.

Recombinant mussel adhesive protein Mgfp-5 as cell adhesion biomaterial

Hwang, Dong Soo,Gim, Youngsoo,Kang, Dong Gyun,Kim, Yeon Kyu,Cha, Hyung Joon Elsevier 2007 Journal of biotechnology Vol.127 No.4

<P><B>Abstract</B></P><P><I>Mytilus galloprovincialis</I> foot protein type-5 (Mgfp-5) is one of the mussel adhesive proteins that participate in adhesion with the substratum. We previously reported the production of recombinant Mgfp-5 in <I>Escherichia coli</I> and showed that the recombinant protein had superior adhesion abilities versus those of Cell-Tak, a commercially available mussel adhesive protein mixture. In the present work, we investigated the feasibility of using recombinant Mgfp-5 as a cell adhesion agent. Purified and tyrosinase-modified recombinant Mgfp-5 was used to adhere living anchorage-independent cells such as insect <I>Drosophila</I> S2 cells and human MOLT-4 cells onto glass slides. Our results revealed that these cell lines efficiently attached to recombinant Mgfp-5-coated glass surfaces, and that surface-immobilized S2 cells were viable and able to undergo cell division for up to 1 week. Cytochemical studies with 4′,6-diamidino-2-phenylindole (DAPI) staining of nuclei and immunofluorescence for secreted foreign human erythropoietin (hEPO) from recombinant S2 cells and quantitative comparative analyses of S2 cell binding ability with Cell-Tak and poly-<SMALL>L</SMALL>-lysine, the main cell adhesion agent, were performed to demonstrate successful usage of recombinant Mgfp-5 for cell biological applications. Collectively, these results indicate that recombinant Mgfp-5 may be a useful new cell adhesion biomaterial for anchorage-independent cells.</P>

Butyrate-induced differentiation of PC12 cells to chromaffin cells involves cell adhesion and induction of extracellular proteins and cell adhesion proteins

Heo, Jee-In,Oh, Soo-Jin,Kho, Yoon-Jung,Kim, Jeong-Hyeon,Kang, Hong-Joon,Park, Seong-Hoon,Kim, Hyun-Seok,Shin, Jong-Yeon,Lee, Sung-Young,Kim, Min-Ju,Min, Bon-Hong,Kim, Sung-Chan,Park, Jae-Bong,Kim, Jae The Korean Society for Integrative Biology 2010 Animal cells and systems Vol.14 No.4

PC12 cells were differentiated into the cells of chromaffin phenotype by butyrate treatment. Cells were aggregated and formed tight cell adhesion. To investigate the molecular change in this differentiation, we examined expression levels of cell adhesion proteins and extracellular proteins during butyrate induced-differentiation of PC12 cells. Integrin ${\beta}1$, integrin ${\alpha}7$, E cadherin, VCAM, collagen-I, fibronectin, desmoglein and connexin were increased during differentiation. The levels of clusterin and secreted clusterin were also increased. These increased levels of cell adhesion proteins and extracellular proteins appear to induce cell aggregation and tight cell adhesion. The levels of p21, p27 and p16 were increased probably because of differentiation-related growth arrest during differentiation. Prolonged incubation of butyrate up to 1 day was required for differentiation. Signal transduction pathways for this differentiatiom could not be identified since various inhibitors had no effect. The results showed that butyrateinduced differentiation of PC12 cells to chromaffin cells involves tight cell adhesion and induction of extracellular proteins and cell adhesion proteins.

BAY11-7082에 의한 U937 세포의 CD29-매개성 세포간 유착과정 조절 효과

김병훈,조재열,Kim, Byung-Hun,Cho, Jae-Youl 대한약학회 2008 약학회지 Vol.52 No.5

BAY11-7082 was initially found to be an anti-inflammatory drug with NF-${\kappa}B$ inhibitory property. In this study, we evaluated modulatory function of BAY11-7082 on U937 cell-cell adhesion induced by CD29 (${\beta}1$-integrins). BAY11-7082 strongly blocked functional activation of CD29 (${\beta}1$-integrins), as assessed by cell-cell adhesion assay. However, this compound did not block a simple activation of CD29, as assessed by cell-fibronectin adhesion assay. In particular, to understand molecular mechanism of BAY11-7082-mediated inhibition, the regulatory roles of CD29-induced actin cytoskeleton rearrangement under cell-cell adhesion and surface level of CD29 were examined using confocal and flow cytometic analysis. Interestingly, this compound strongly suppressed the molecular association of actin cytoskeleton with CD29 at cell-cell adhesion site. Moreover, BAY11-7082 also diminished surface levels of CD29 as well as its-associated adhesion molecule CD147, but not other adhesion molecules such as CD18 and CD43. Therefore, our data suggest that BAY11-7082 may be involved in regulating immune responses managed by CD29-mediated cell-cell adhesion.

단핵구 기능 수행에서의 $CD29({\beta}1-integrins)$ 조절 역할

김병훈,조재열,Kim, Byung-Hun,Cho, Jae-Youl 대한약학회 2008 약학회지 Vol.52 No.1

CD29 $({\beta}1-integrins)$ is one of major adhesion molecules involved in regulating cell adhesion, migration and morphological changes. In this study, we investigated the regulatory role of CD29 in monocytic functions using monocytic cell line U937 cells. CD29 was found to be one of highly expressed membrane proteins in U937 cells, according to flow cytometric analysis. The activation of CD29 by agonistic antibody MEM101A and extracellular matrix protein (ECM) fibronectin strongly induced cell-cell and cell-fibronectin adhesions. However, blocking antibodies to CD98 and CD147 showed different inhibitory features in these two adhesion events. Furthermore, U0126, an ERK inhibitor, only blocked cell-cell adhesion but not cell-fibronectin adhesion, indicating that cell-cell or cell-fibronectin adhesion events may be regulated by different molecular mechanisms. Meanwhile, CD29 activation also enhanced ROS generation but not phagocytic ability, and similarly radical scavenger N-acetyl-L-cysteine strongly blocked CD29-mediated cell-cell adhesion, implying that ROS may play a critical role in up-regulating cell-cell adhesion. Therefore, our data suggest that the activation of CD29 may be critically involved in regulating monocytic cell-mediated cell-cell adhesion and ROS generation.

RANKL stimulates proliferation, adhesion and IL-7 expression of thymic epithelial cells

Hee-Woo Lee,Hye Kyung Park,Yong Jin Na,Chi Dae Kim,Jung Hoon Lee,Bong Seon Kim,Jae Bong Kim,Choong Won Lee,Jeon Ok Moon,Sik Yoon 생화학분자생물학회 2008 Experimental and molecular medicine Vol.40 No.1

In many clinical situations which cause thymic in- volution and thereby result in immune deficiency,T cells are the most often affected,leading to a prolonged deficiency of T cells.Since only the thymic-dependent T cell production pathway secures stable regeneration of fully mature T cells,seeking strategies to enhance thymic regeneration should be a key step in developing therapeutic methods for the treatment of these sig- nificant clinical problems.This study clearly shows that receptor activator of NF-κB ligand (RANKL) stim- ulates mouse thymic epithelial cell activities including cell proliferation,thymocyte adhesion to thymic epi-thelial cells, and the expression of cell death regulatory genes favoring cell survival, cell adhesion molecules such as ICAM-1 and VCAM-1, and thymopoietic factors including IL-7. Importantly, RANKL exhibited a sig- nificant capability to facilitate thymic regeneration in mice.In addition,this study demonstrates that RANKL acts directly on the thymus to activate thymus re- generation regardless of its potential influences on thymic regeneration through an indirect or systemic effect.In light of this,the present study provides a greater insight into the development of novel ther- apeutic strategies for effective thymus repopulation using RANKL in the design of therapies for many clin- ical conditions in which immune reconstitution is required. In many clinical situations which cause thymic in- volution and thereby result in immune deficiency,T cells are the most often affected,leading to a prolonged deficiency of T cells.Since only the thymic-dependent T cell production pathway secures stable regeneration of fully mature T cells,seeking strategies to enhance thymic regeneration should be a key step in developing therapeutic methods for the treatment of these sig- nificant clinical problems.This study clearly shows that receptor activator of NF-κB ligand (RANKL) stim- ulates mouse thymic epithelial cell activities including cell proliferation,thymocyte adhesion to thymic epi-thelial cells, and the expression of cell death regulatory genes favoring cell survival, cell adhesion molecules such as ICAM-1 and VCAM-1, and thymopoietic factors including IL-7. Importantly, RANKL exhibited a sig- nificant capability to facilitate thymic regeneration in mice.In addition,this study demonstrates that RANKL acts directly on the thymus to activate thymus re- generation regardless of its potential influences on thymic regeneration through an indirect or systemic effect.In light of this,the present study provides a greater insight into the development of novel ther- apeutic strategies for effective thymus repopulation using RANKL in the design of therapies for many clin- ical conditions in which immune reconstitution is required.

EFFECT OF TITANIUM SURFACE ROUGHNESS ON CELL ADHESION OF HUMAN OSTEOBLAST-LIKE CELLS (MG63)

Yim Soon-Ho The Korean Academy of Prosthodonitics 2004 대한치과보철학회지 Vol.42 No.3

Statement of problem. The effects of surface roughness have not or insufficiently been analyzed on earlier events such as cell adhesion though cell behavior most germane to implant performance is cell adhesion. Purpose. The purpose of this study was to evaluate cell adhesion of osteoblast-like cells (MG63) onto three types of titanium disks with varying roughness using the Elisa assay. Materials and methods. Representative disks from each group (SLA, HA, machined) were subjected to surface analysis and surface roughness was measured by the optical interferometer (Accura 2000, Intekplus Co., Seoul, Korea). Following this, MG63 cells were cultured on the titanium disks and released. Cell adhesion measurements using the Elisa assay were performed specifically at three points: after 24, 48, and 72 hours of culture. Results. Among the 3 types of surface analyzed, the SLA surface was the roughest with a Ra value of $1.114{\mu}m$ followed by HA coated surface and machined surface, consecutively. The optical density values for the SLA surface group was significantly higher than that of the machined and HA coated surface groups following 24 and 48 hours of culture. The cell culture on HA coated surface showed significantly higher values compared to the machined surface following 24, 48 and 72 hours of culture. Conclusion. The results suggest that surface treatment of titanium surfaces enhanced cell adhesion of human osteoblast-like cells (MG63).

Effect of Various Factors on Early THP-1 Cell Adhesion Induced Phorbol 12-Myristate 13-Acetate (PMA)

Yong Sam Jo(조용삼),Ji Hyun Shin(신지현),Tae Saeng Choi(최태생) 한국생명과학회 2008 생명과학회지 Vol.18 No.7

본 실험에서는 THP-1 세포의 PMA에 의하여 유도되는 초기 세포부착에 관한 메카니즘을 이해하기 위하여 다양한 요인(혈청, 신규 단백질의 합성, 세포 골격 저해제, 단백질 인신화 저해제)들의 효과를 조사하였다. 또한 본 실험에서는 이들 세포부착의 정도를 일반적으로 세포증식 분석에 사용되고 있는 SRB염색법을 도입하여 세포부착 분석에 간편한 방법의 조건을 확립하였다. PMA에 의한 초기 세포부착에는 배양액중의 혈청의 유무는 영향이 없었으나, 신규 단백질의 합성이 요구되는 것을 확인하였다. 또한 이들 초기 세포부착에 PMA처리에 의한 PKC의 활성화는 필수적이나, 그 하류 활성화 인자로 잘 알려진 MAP-kinase (erk1/2)의 인산화는 필요치 않음을 알 수 있었다. 흥미롭게도 액틴 중합 저해제인 cytochalasin D의 PMA와 공 처리는 오히려 세포부착을 PMA 단독 처리시 보다 증가시켰다. 또한 본 실험에서 사용된 SRB 염색법을 통한 세포부착 분석법은 최근 암 등 다양한 질환의 신약 표적 분자로 주목을 받고 있는 PKC 저해제의 초기 세포 기반 분석에 매우 유용하리라고 생각된다. We evaluated the effects of various factors (e.g., serum, inhibitors of protein synthesis, and cytoskeleton and protein kinases) on early PMA-induced THP-1 cell adhesion using an adhesion assay with Sulforhodamine B (SRB) staining, which was used to assess the proliferation of the attached cells. THP-1 cell adhesion to a plastic substrate was detected 1 hr after exposure to Phorbol 12-Myristate 13-Acetate (PMA) and peaked after 18 hr. At concentrations > 25 nM PMA, the level of adhesion did not change. Based on our preliminary results, we used 25 nM PMA and 5 hr of culture as standard assay conditions. Early PMA-induced cell adhesion was not affected by the presence of serum or PD 98059 in the culture medium, but was affected by the addition of PKC inhibitors and cycloheximide. In the presence of actin inhibitor with PMA, the cell adhesion increased when comparing with PMA treatment only. Thus, early PMA-induced adhesion of THP-1 cells does not require serum in the culture medium, MAP-kinase activation, or actin polymerization, but does require de novo protein synthesis and PKC activation. Our SRB-based cell adhesion assay may be used to screen other PKC inhibitors.

Interferon-gamma, Tumor Necrosis Factor-alpha및 Transforming Growth Factor-beta(1)에 의한 골수세포의 유착분자 발현 및 Stromal Cell-Derived Factor-1 생성의 조절

조덕연,황진희,정효균,박수진,박상은,곽승근,윤환중,성주명,김삼용 大韓血液學會 2003 大韓血液學會誌 Vol.38 No.2