[Focal adhesion kinase and mitogen activated protein kinase are involved in vascular smooth muscle cells migration and proliferation stimulated by fibronectin].
OBJECTIVE
To study the roles of focal adhesion kinase (FAK) and mitogen activated protein kinase (p42/44MAPK) in the process of migration and proliferation of vascular smooth muscle cells (VSMC) stimulated by fibronectin (FN).


METHODS
VSMCs were taken from the tunica media of SD rats and cultured. Migration and proliferation of cultured VSMCs were stimulated by different concentrations of FN; FAK, p42/44MAPK and their phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense oligodeoxynucleotide (ODN) was transfected into VSMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation. VSMC migration and proliferation were also measured by modified Boyden Chamber and (3)H-thymidine respectively.


RESULTS
FAK and p42/44MAPK were expressed when VSMC adhesion and migration were successfully simulated by FN (5, 10, 20, 40, and 60 microg/ml), high contents of FAK and p42/44MAPK phosphorylation were detected in groups with 20 microgram/ml FN or more. FAK antisense ODN was transfected efficiently by cationic lipid. Phosphorylation of FAK and p42/44MAPK was inhibited significantly after FAK antisense ODN transfection. Cell migration stimulated by FN of the concentrations of 10, 20, 40, and 60 microg/ml was reduced by 23.26%, 21.63%, 19.31% and 17.88% respectively (P < 0.05). VSMC proliferation in 5 approximately 60 microgram/ml FN groups was reduced by 27.67% approximately 46.67% (P < 0.05).


CONCLUSIONS
FAK and p42/44MAPK play an important role in VSMC migration and proliferation stimulated by extracellular matrix. The process can be inhibited by FAK antisense ODN effectively.
Department of Cardiology, First Hospital, Peking University, Beijing 100034, China.
Hang
H
Yin
Lihui
L
Wang
Yong
Y
Huo
Xu
X
Peng
Chunfang
C
Xia
Chaoshu
C
Tang
D000818:Animals; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D005353:Fibronectins / Q000494:pharmacology*; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D066298:In Vitro Techniques; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D005353:Fibronectins; D020319:Oligodeoxyribonucleotides, Antisense; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496052:Ptk2 protein, rat; D020928:Mitogen-Activated Protein Kinases