Evaluation of the correlation between focal adhesion kinase phosphorylation and cell adhesion force using "DEP" technology.
Dielectrophoresis (DEP) is the phenomenon in which a particle, such as a living cell, is polarized and moved by electrical gravity in a non-uniform electric field. In the present study, the DEP force is utilized to act on the cells to induce spatial movement for investigating the correlation between the cell adhesion force and activation level of focal adhesion kinase (FAK). The DEP force produced by the non-uniform electric field was used to measure the cell adhesion force of ECV304 cells, on type 1 collagen (COL1)- and fibronectin (FN)-coated polydimethylsiloxane (PDMS) membranes. For COL1-coating, ECV304 cells revealed weak and variable adhesion force (0.343-0.760 nN) in the first eight hours of incubation. Interestingly, the cell adhesion force of ECV304 at two and five hours of cultivation was significantly high and matched their FAK activation level. In comparison, ECV304 on FN-coated membrane had higher and more stable cell adhesion force (0.577-2.053 nN). FN coating intensified the cell adhesion force of ECV304 with culture time and similar outcome was present on the activation level of FAK. Therefore, this study demonstrated a relationship between cell adhesion force and FAK activation level that was dependent on the choice of the extracellular matrix (ECM) component. Subsequently, two tyrosine kinase inhibitors (AG18 and genistein) and one PI3K inhibitor (LY294002) were applied to study the influence of protein phosphorylation on the cell adhesion force. FAK plays an important role on cell attachment and DEP force measurement is a useful technique for studying cell adhesion.
Department of Biomechatronic Engineering, National Chiayi University, No. 300, University Road, East District, Chiayi 600, Taiwan. cay@mail.ncyu.edu.tw
Chyung
C
Ay
Chih-Chang
CC
Yeh
Min-Chih
MC
Hsu
Huaang-Youh
HY
Hurng
Philip Chi Lip
PC
Kwok
Hsin-I
HI
Chang
D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004586:Electrophoresis; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D001743:Urinary Bladder / Q000166:cytology / Q000201:enzymology
D004129:Dimethylpolysiloxanes; D047428:Protein Kinase Inhibitors; C013830:baysilon; D051416:Focal Adhesion Protein-Tyrosine Kinases