Fibrillar collagen specifically regulates human vascular smooth muscle cell genes involved in cellular responses and the pericellular matrix environment.
Proliferation and alpha(v)beta(3) integrin-dependent migration of vascular smooth muscle cells are suppressed on polymerized type I collagen. To identify genes specifically regulated in human smooth muscle cells by polymerized collagen, we used the suppressive subtraction hybridization technique. Compared with smooth muscle cells cultured on monomer collagen, polymerized collagen suppresses the following: (1) a number of other extracellular matrix proteins, including fibronectin, thrombospondin-1, tenascin-C, and cysteine-rich protein 61; (2) actin binding proteins including alpha-actinin; (3) signaling molecules; (4) protein synthesis-associated proteins; and (5) genes with unknown functions. Some of the identified genes, including cysteine-rich protein 61, show unique kinetics of mRNA regulation by monomer or polymerized collagen distinct from growth factors, suggesting extracellular matrix-specific gene modulation. Moreover, in vivo balloon catheter-mediated injury to the rat carotid artery induces many of the genes that are suppressed by polymerized collagen. Protein levels of thrombospondin-1 and fibronectin are also suppressed by polymerized collagen. Thrombospondin-1-mediated smooth muscle cell migration on vitronectin is significantly inhibited after culture on polymerized collagen for 24 hours, which is associated with decreased alpha-actinin accumulation at focal adhesions. Thus, polymerized type I collagen dynamically regulates gene expression, pericellular accumulation of extracellular matrix molecules, and the response to a given matrix molecule.
Ichii
T
T
Department of Biochemistry, Internal Medicine, Osaka City University Medical School, Osaka, Japan.
Koyama
H
H
Tanaka
S
S
Kim
S
S
Shioi
A
A
Okuno
Y
Y
Raines
E W
EW
Iwao
H
H
Otani
S
S
Nishizawa
Y
Y
D000185:Actinin / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D015152:Blotting, Northern; D002339:Carotid Arteries / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D020212:Carotid Artery Injuries / Q000209:etiology / Q000235:genetics; D002404:Catheterization; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D002633:Chemotaxis / Q000187:drug effects; D003094:Collagen / Q000737:chemistry / Q000494:pharmacology*; D018076:DNA, Complementary / Q000235:genetics; D004195:Disease Models, Animal; D016326:Extracellular Matrix Proteins / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006133:Growth Substances / Q000494:pharmacology; D006801:Humans; D016023:Integrins / Q000502:physiology; D007700:Kinetics; D008297:Male; D018613:Microscopy, Confocal; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D009693:Nucleic Acid Hybridization / Q000379:methods; D011108:Polymers; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D019700:Thrombospondin 1 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D019096:Vitronectin / Q000494:pharmacology
D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D006133:Growth Substances; D016023:Integrins; D011108:Polymers; D012333:RNA, Messenger; D019700:Thrombospondin 1; D019096:Vitronectin; D000185:Actinin; D003094:Collagen