title: Inhibition of versican expression by siRNA facilitates tropoelastin synthesis and elastic fiber formation by human SK-LMS-1 leiomyosarcoma smooth muscle cells in vitro and in vivo.
abstract: Versican is an extracellular matrix (ECM) molecule that interacts with other ECM components to influence ECM organization, stability, composition, and cell behavior. Versican is known to increase in a number of cancers, but little is known about how versican influences the amount and organization of the ECM components in the tumor microenvironment. In the present study, we modulated versican expression using siRNAs in the human leiomyosarcoma (LMS) smooth muscle cell line SK-LMS-1, and observed the formation of elastin and elastic fibers in vitro and also in vivo in a nude mouse tumor model. Constitutive siRNA-directed knockdown of versican in LMS cells resulted in increased levels of elastin, as shown by immunohistochemical staining of the cells in vitro, and by mRNA and protein analyses. Moreover, versican siRNA LMS cells, when injected into nude mice, generated smaller tumors that had significantly greater immunohistochemical and histochemical staining for elastin when compared to control tumors. Additionally, microarray analyses were used to determine the influence of versican isoform modulation on gene expression profiles, and to identify genes that influence and relate to the process of elastogenesis. cDNA microarray analysis and TaqMan low density array validation identified previously unreported genes associated with downregulation of versican and increased elastogenesis. These results highlight an important role for the proteoglycan versican in regulating the expression and assembly of elastin and the phenotype of LMS cells.
authors.affiliation: Matrix Biology Program, Benaroya Research Institute, 1201 Ninth Avenue, Seattle, WA 98101, USA; Department of Pathology, University of Washington, Seattle, WA 98195, USA.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Keire
authors.affiliation: Matrix Biology Program, Benaroya Research Institute, 1201 Ninth Avenue, Seattle, WA 98101, USA.
authors.forename: Steven L
authors.initials: SL
authors.lastname: Bressler
authors.affiliation: Department of Pathology, University of Washington, Seattle, WA 98195, USA.
authors.forename: Eileen R
authors.initials: ER
authors.lastname: Mulvihill
authors.affiliation: Department of Biochemistry, University of Texas Health Science Center at Tyler, Tyler, TX 75708, USA.
authors.forename: Barry C
authors.initials: BC
authors.lastname: Starcher
authors.affiliation: Matrix Biology Program, Benaroya Research Institute, 1201 Ninth Avenue, Seattle, WA 98101, USA.
authors.forename: Inkyung
authors.initials: I
authors.lastname: Kang
authors.affiliation: Matrix Biology Program, Benaroya Research Institute, 1201 Ninth Avenue, Seattle, WA 98101, USA; Department of Pathology, University of Washington, Seattle, WA 98195, USA. Electronic address: twight@benaroyaresearch.org.
authors.forename: Thomas N
authors.initials: TN
authors.lastname: Wight
mesh_terms: D000818:Animals; D002460:Cell Line; D004547:Elastic Tissue / Q000378:metabolism / Q000473:pathology*; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D006801:Humans; D066298:In Vitro Techniques; D007890:Leiomyosarcoma / Q000235:genetics / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D034741:RNA, Small Interfering / Q000378:metabolism*; D014333:Tropoelastin / Q000096:biosynthesis*; D053675:Versicans / Q000235:genetics* / Q000378:metabolism
chemical_list: D034741:RNA, Small Interfering; D014333:Tropoelastin; C508322:VCAN protein, human; C508323:Vcan protein, mouse; D053675:Versicans
keywords: Elastin; Elastogenesis; Extracellular matrix; Proliferation; SiRNA; Versican