J Korean Orthop Res Soc.  2001 Oct;4(2):159-166.

The control of chondroid cell's adhesiveness by modulation of focal adhesion kinase(FAK) expression

• Lee JW
• Kim YH
• Hahn SB
• Shin KH
• Park HB

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.

Keyword

Chondroid cell; cell attachment; focal adhesion kinase; integrin-beta1; FAK-CD; adenovirus vector

MeSH Terms

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
Antibodies
Biocompatible Materials
Collagen Type II
Focal Adhesion Protein-Tyrosine Kinases
Peptides
RNA, Messenger
Tyrosine