Focal adhesion kinase(FAK)의 발현 조절에 의한 연골양 세포 부착의 조절
The control of chondroid cell's adhesiveness by modulation of focal adhesion kinase(FAK) expression
이진우 ; 김윤희 ; 박희붕 ; 신규호 ; 한수봉
Journal of Korean Orthopaedic Research Society (대한정형외과연구학회지), Vol.4(2) : 159~166, 2001
Journal of Korean Orthopaedic Research Society (대한정형외과연구학회지)
Purpose : We propose that cell attachment can be regulated by the modulation of FAK expression using an adenovirus vector. Materials and Methods : Chondrocytes and chondroid cells were used in cell attachment test by blocking or non-blocking of antibodies and synthetic peptides on type II collagen precoated 96-well immunoplates. The Cterminal domain of FAK(FAK-CD) was transfected through infection of the recombinant adenovirus. Also tyrosine phosphorylation of FAK was checked by immunoprecipitation of FAK followed by western blot analysis with anti-phosphotyrosine antibody. For evaluating the change of integrin expression, semi-quantitative reverse-transcription polymerase chain(RT-PCR) reactions were done after transfection of FAK-CD. Results : We observed more increased expression of FAK in the chondroid cells than that in chondrocytes using western blotting. The level of attachment to type II collagen was significantly inhibited by blocking with the monoclonal antibody of integrin-β1 and synthetic RGD peptides. Also adenovirus mediated transfection of FAK-CD resulted in inhibition of phosphorylation of FAK and significantly inhibited cell attachment in only JJ102. Integrin-β1 antibody blocking after transfection with FAK-CD showed inhibition of cell attachment in more than 95% of all cells. The mRNA expression of both Integrin 2 and integrin 5 was increased but was not significant. Protein expression of integrin 2 and integrin 5 showed no changes. Conclusion : We found that the attachment of FAK-overexpressing cells could be mediated through integrin-β1 receptor. We concluded that the modification of FAK expression will contribute to increase the cell attachment to biomaterials and regeneration of cartilage defects.