The control of chondroid cell's adhesiveness by modulation of focal adhesion kinase(FAK) expression.
- Author:
Jin Woo LEE
1
;
Yun Hee KIM
;
Soo Bong HAHN
;
Kyu Ho SHIN
;
Hee Boong PARK
Author Information
1. Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea. ljwos@yumc.yonsei.ac.kr
- Publication Type:Original Article
- Keywords:
Chondroid cell;
cell attachment;
focal adhesion kinase;
integrin-beta1;
FAK-CD;
adenovirus vector
- MeSH:
Adenoviridae;
Adhesiveness*;
Antibodies;
Biocompatible Materials;
Blotting, Western;
Cartilage;
Chondrocytes;
Collagen Type II;
Focal Adhesion Protein-Tyrosine Kinases;
Focal Adhesions*;
Immunoprecipitation;
Peptides;
Phosphorylation;
Regeneration;
RNA, Messenger;
Transfection;
Tyrosine
- From:Journal of Korean Orthopaedic Research Society
2001;4(2):159-166
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
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 C-terminal 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-beta1 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-beta1 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 a2 and integrin a5 was increased but was not significant. Protein expression of integrin a2 and integrin a5 showed no changes. CONCLUSION: We found that the attachment of FAK-overexpressing cells could be mediated through integrin-beta1 receptor. We concluded that the modification of FAK expression will contribute to increase the cell attachment to biomaterials and regeneration of cartilage defects.
