title: Clusterin regulates vascular smooth muscle cell nodule formation and migration.
abstract: Vascular smooth muscle cells (VSMC) are the principal cellular component of the blood vessel wall where they exist in a differentiated state to maintain vascular tone. However, VSMC are not terminally differentiated and can be induced to dediffentiate, proliferate, and migrate. In fact, smooth muscle cell migration from the vascular wall into the lumen of the vessel is a central feature of occlusive vascular pathologies including atherosclerosis and intimal hyperplasia. In vitro, in the presence of an extracellular matrix, cultured vascular smooth muscle cells can migrate and invade the underlying gelatinous matrix, form multicellular nodular aggregations, and secrete the glycoprotein clusterin. Nodular cultures appear to mimic some of the properties of differentiated VSMC, in vivo. Here, to test the hypothesis that clusterin functions to modulate the formation of VSMC nodules and to facilitate cell migration a clusterin negative VSMC clone, SM-CLU13AS (Moulson and Millis, 1999, J Cell Physiol 180:355), was transiently transfected with plasmid pRcCMVCLU that contains the full-length porcine clusterin cDNA sequence under control of the CMV promoter. The transiently transfected VSMC culture expressed and secreted clusterin and formed nodules. To determine if clusterin regulates VSMC migration we used modified Boyden chamber assays. Clusterin, at 10 microg/ml, clearly promotes VSMC migration. In addition, a 15 amino acid synthetic peptide, representing amino acids 118-132 [KQTCMKFYARVCRSG] of the mature clusterin polypeptide, inhibits VSMC attachment to gelatinous substrate. Finally, clusterin appears to have a role in regulating endogenous clusterin expression in the clusterin negative clone. These results clearly establish that clusterin has functional role in VSMC nodule formation and support the conclusion that clusterin is a critical component of smooth muscle cell phenotypic modulation.
authors.affiliation: Center for Comparative Functional Genomics, Department of Biological Sciences, University at Albany-SUNY, 1400 Washington Avenue, Albany, NY 12222, USA. millis@csc.albany.edu
authors.forename: A J
authors.initials: AJ
authors.lastname: Millis
authors.forename: M
authors.initials: M
authors.lastname: Luciani
authors.forename: H M
authors.initials: HM
authors.lastname: McCue
authors.forename: M E
authors.initials: ME
authors.lastname: Rosenberg
authors.forename: C L
authors.initials: CL
authors.lastname: Moulson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001013:Aorta, Thoracic / Q000166:cytology / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology*; D002455:Cell Division; D002465:Cell Movement / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002999:Clone Cells; D051152:Clusterin; D017077:Culture Media, Conditioned; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D051379:Mice; D018832:Molecular Chaperones / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000502:physiology*; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism; D013552:Swine; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: C495243:Clu protein, mouse; D051152:Clusterin; D017077:Culture Media, Conditioned; D006023:Glycoproteins; D018832:Molecular Chaperones; D010446:Peptide Fragments; D011994:Recombinant Proteins