Search tips
Search criteria 


Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. 1993 September 2; 122(6): 1285–1294.
PMCID: PMC2119864

Suppression of vinculin expression by antisense transfection confers changes in cell morphology, motility, and anchorage-dependent growth of 3T3 cells


The expression of vinculin, a major component of adhesion plaques and cell-cell junctions, is markedly modulated in cells during growth activation, differentiation, motility and cell transformation. The stimulation of quiescent cells by serum factors and the culturing of cells on highly adhesive matrices induce vinculin gene expression, whereas the transformation of fibroblast and epithelial cells often results in decreased vinculin expression (reviewed in Rodriguez Fernandez, J. L., B. Geiger, D. Salomon, I. Sabanay, M. Zoller, and A. Ben-Ze'ev. 1992. J. Cell Biol. 119:427). To study the effect of reduced vinculin expression on cell behavior, 3T3 cells were transfected with an antisense vinculin cDNA construct, and clones displaying decreased vinculin levels down to 10-30% of control levels were isolated. These cells showed a round phenotype with smaller and fewer vinculin-positive plaques localized mostly at the cell periphery. In addition, they displayed an increased motility compared to controls, manifested by a faster closure of "wounds" introduced into the monolayer, and by the formation of longer phagokinetic tracks. Moreover, the antisense transfectants acquired a higher cloning efficiency and produced larger colonies in soft agar than the parental counterparts. The results demonstrate that the regulation of vinculin expression in cells can affect, in a major way, cell shape and motility, and that decreased vinculin expression can induce cellular changes reminiscent of those found in transformed cells.

Full Text

The Full Text of this article is available as a PDF (3.7M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Barrandon Y, Green H. Cell migration is essential for sustained growth of keratinocyte colonies: the roles of transforming growth factor-alpha and epidermal growth factor. Cell. 1987 Sep 25;50(7):1131–1137. [PubMed]
  • Bellas RE, Bendori R, Farmer SR. Epidermal growth factor activation of vinculin and beta 1-integrin gene transcription in quiescent Swiss 3T3 cells. Regulation through a protein kinase C-independent pathway. J Biol Chem. 1991 Jun 25;266(18):12008–12014. [PubMed]
  • Bendori R, Salomon D, Geiger B. Contact-dependent regulation of vinculin expression in cultured fibroblasts: a study with vinculin-specific cDNA probes. EMBO J. 1987 Oct;6(10):2897–2905. [PubMed]
  • Bendori R, Salomon D, Geiger B. Identification of two distinct functional domains on vinculin involved in its association with focal contacts. J Cell Biol. 1989 Jun;108(6):2383–2393. [PMC free article] [PubMed]
  • Ben-Ze'ev A. Application of two-dimensional gel electrophoresis in the study of cytoskeletal protein regulation during growth activation and differentiation. Electrophoresis. 1990 Mar;11(3):191–200. [PubMed]
  • Ben-Ze'ev A. Animal cell shape changes and gene expression. Bioessays. 1991 May;13(5):207–212. [PubMed]
  • Ben-Ze'ev A. Cytoarchitecture and signal transduction. Crit Rev Eukaryot Gene Expr. 1992;2(3):265–281. [PubMed]
  • Ben-Ze'ev A, Amsterdam A. In vitro regulation of granulosa cell differentiation. Involvement of cytoskeletal protein expression. J Biol Chem. 1987 Apr 15;262(11):5366–5376. [PubMed]
  • Ben-Ze'ev A, Farmer SR, Penman S. Protein synthesis requires cell-surface contact while nuclear events respond to cell shape in anchorage-dependent fibroblasts. Cell. 1980 Sep;21(2):365–372. [PubMed]
  • Ben-Ze'ev A, Reiss R, Bendori R, Gorodecki B. Transient induction of vinculin gene expression in 3T3 fibroblasts stimulated by serum-growth factors. Cell Regul. 1990 Aug;1(9):621–636. [PMC free article] [PubMed]
  • Burridge K, Fath K, Kelly T, Nuckolls G, Turner C. Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Biol. 1988;4:487–525. [PubMed]
  • Burridge K, Turner CE, Romer LH. Tyrosine phosphorylation of paxillin and pp125FAK accompanies cell adhesion to extracellular matrix: a role in cytoskeletal assembly. J Cell Biol. 1992 Nov;119(4):893–903. [PMC free article] [PubMed]
  • Cunningham CC, Stossel TP, Kwiatkowski DJ. Enhanced motility in NIH 3T3 fibroblasts that overexpress gelsolin. Science. 1991 Mar 8;251(4998):1233–1236. [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Coutu MD, Craig SW. cDNA-derived sequence of chicken embryo vinculin. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8535–8539. [PubMed]
  • Geiger B. A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells. Cell. 1979 Sep;18(1):193–205. [PubMed]
  • Geiger B. Cytoskeleton-associated cell contacts. Curr Opin Cell Biol. 1989 Feb;1(1):103–109. [PubMed]
  • Geiger B, Ayalon O, Ginsberg D, Volberg T, Rodríguez Fernández JL, Yarden Y, Ben-Ze'ev A. Cytoplasmic control of cell adhesion. Cold Spring Harb Symp Quant Biol. 1992;57:631–642. [PubMed]
  • Geiger B, Ginsberg D, Salomon D, Volberg T. The molecular basis for the assembly and modulation of adherens-type junctions. Cell Differ Dev. 1990 Dec 2;32(3):343–353. [PubMed]
  • Geiger B, Salomon D, Takeichi M, Hynes RO. A chimeric N-cadherin/beta 1-integrin receptor which localizes to both cell-cell and cell-matrix adhesions. J Cell Sci. 1992 Dec;103(Pt 4):943–951. [PubMed]
  • Glück U, Rodríguez Fernández JL, Pankov R, Ben-Ze'ev A. Regulation of adherens junction protein expression in growth-activated 3T3 cells and in regenerating liver. Exp Cell Res. 1992 Oct;202(2):477–486. [PubMed]
  • Glück U, Kwiatkowski DJ, Ben-Ze'ev A. Suppression of tumorigenicity in simian virus 40-transformed 3T3 cells transfected with alpha-actinin cDNA. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):383–387. [PubMed]
  • Guan JL, Shalloway D. Regulation of focal adhesion-associated protein tyrosine kinase by both cellular adhesion and oncogenic transformation. Nature. 1992 Aug 20;358(6388):690–692. [PubMed]
  • Heckman CA, Oravecz KI, Schwab D, Pontén J. Ruffling and locomotion: role in cell resistance to growth factor-induced proliferation. J Cell Physiol. 1993 Mar;154(3):554–565. [PubMed]
  • Inouye M. Antisense RNA: its functions and applications in gene regulation--a review. Gene. 1988 Dec 10;72(1-2):25–34. [PubMed]
  • Izant JG. Antisense "pseudogenetics". Cell Motil Cytoskeleton. 1989;14(1):81–91. [PubMed]
  • Izant JG, Weintraub H. Inhibition of thymidine kinase gene expression by anti-sense RNA: a molecular approach to genetic analysis. Cell. 1984 Apr;36(4):1007–1015. [PubMed]
  • Izant JG, Weintraub H. Constitutive and conditional suppression of exogenous and endogenous genes by anti-sense RNA. Science. 1985 Jul 26;229(4711):345–352. [PubMed]
  • Jones P, Jackson P, Price GJ, Patel B, Ohanion V, Lear AL, Critchley DR. Identification of a talin binding site in the cytoskeletal protein vinculin. J Cell Biol. 1989 Dec;109(6 Pt 1):2917–2927. [PMC free article] [PubMed]
  • Juliano RL, Haskill S. Signal transduction from the extracellular matrix. J Cell Biol. 1993 Feb;120(3):577–585. [PMC free article] [PubMed]
  • Kim SK, Wold BJ. Stable reduction of thymidine kinase activity in cells expressing high levels of anti-sense RNA. Cell. 1985 Aug;42(1):129–138. [PubMed]
  • Knecht DA, Loomis WF. Antisense RNA inactivation of myosin heavy chain gene expression in Dictyostelium discoideum. Science. 1987 May 29;236(4805):1081–1086. [PubMed]
  • Knecht D. Application of antisense RNA to the study of the cytoskeleton: background, principles, and a summary of results obtained with myosin heavy chain. Cell Motil Cytoskeleton. 1989;14(1):92–102. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • LaFlamme SE, Akiyama SK, Yamada KM. Regulation of fibronectin receptor distribution. J Cell Biol. 1992 Apr;117(2):437–447. [PMC free article] [PubMed]
  • Liu T, Williams JG, Clarke M. Inducible expression of calmodulin antisense RNA in Dictyostelium cells inhibits the completion of cytokinesis. Mol Biol Cell. 1992 Dec;3(12):1403–1413. [PMC free article] [PubMed]
  • Price GJ, Jones P, Davison MD, Patel B, Bendori R, Geiger B, Critchley DR. Primary sequence and domain structure of chicken vinculin. Biochem J. 1989 Apr 15;259(2):453–461. [PubMed]
  • Pollenz RS, Chen TL, Trivinos-Lagos L, Chisholm RL. The Dictyostelium essential light chain is required for myosin function. Cell. 1992 Jun 12;69(6):951–962. [PubMed]
  • Raz A, Zöller M, Ben-Ze'ev Cell configuration and adhesive properties of metastasizing and non-metastasizing BSp73 rat adenocarcinoma cells. Exp Cell Res. 1986 Jan;162(1):127–141. [PubMed]
  • Ridley AJ, Hall A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell. 1992 Aug 7;70(3):389–399. [PubMed]
  • Rodríguez Fernández JL, Ben-Ze'ev A. Regulation of fibronectin, integrin and cytoskeleton expression in differentiating adipocytes: inhibition by extracellular matrix and polylysine. Differentiation. 1989 Dec;42(2):65–74. [PubMed]
  • Rodríguez Fernández JL, Geiger B, Salomon D, Sabanay I, Zöller M, Ben-Ze'ev A. Suppression of tumorigenicity in transformed cells after transfection with vinculin cDNA. J Cell Biol. 1992 Oct;119(2):427–438. [PMC free article] [PubMed]
  • Rodríguez Fernández JL, Geiger B, Salomon D, Ben-Ze'ev A. Overexpression of vinculin suppresses cell motility in BALB/c 3T3 cells. Cell Motil Cytoskeleton. 1992;22(2):127–134. [PubMed]
  • Schwartz MA. Transmembrane signalling by integrins. Trends Cell Biol. 1992 Oct;2(10):304–308. [PubMed]
  • Sefton BM, Hunter T, Ball EH, Singer SJ. Vinculin: a cytoskeletal target of the transforming protein of Rous sarcoma virus. Cell. 1981 Apr;24(1):165–174. [PubMed]
  • Shaulsky G, Goldfinger N, Tosky MS, Levine AJ, Rotter V. Nuclear localization is essential for the activity of p53 protein. Oncogene. 1991 Nov;6(11):2055–2065. [PubMed]
  • Schulze H, Huckriede A, Noegel AA, Schleicher M, Jockusch BM. Alpha-actinin synthesis can be modulated by antisense probes and is autoregulated in non-muscle cells. EMBO J. 1989 Dec 1;8(12):3587–3593. [PubMed]
  • Shin SI, Freedman VH, Risser R, Pollack R. Tumorigenicity of virus-transformed cells in nude mice is correlated specifically with anchorage independent growth in vitro. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4435–4439. [PubMed]
  • Stoker M, O'Neill C, Berryman S, Waxman V. Anchorage and growth regulation in normal and virus-transformed cells. Int J Cancer. 1968 Sep 15;3(5):683–693. [PubMed]
  • Trevor K, Linney E, Oshima RG. Suppression of endo B cytokeratin by its antisense RNA inhibits the normal coexpression of endo A cytokeratin. Proc Natl Acad Sci U S A. 1987 Feb;84(4):1040–1044. [PubMed]
  • Tsukita S, Tsukita S, Nagafuchi A, Yonemura S. Molecular linkage between cadherins and actin filaments in cell-cell adherens junctions. Curr Opin Cell Biol. 1992 Oct;4(5):834–839. [PubMed]
  • Ungar F, Geiger B, Ben-Ze'ev A. Cell contact- and shape-dependent regulation of vinculin synthesis in cultured fibroblasts. Nature. 319(6056):787–791. [PubMed]
  • Volberg T, Geiger B, Dror R, Zick Y. Modulation of intercellular adherens-type junctions and tyrosine phosphorylation of their components in RSV-transformed cultured chick lens cells. Cell Regul. 1991 Feb;2(2):105–120. [PMC free article] [PubMed]
  • Volberg T, Zick Y, Dror R, Sabanay I, Gilon C, Levitzki A, Geiger B. The effect of tyrosine-specific protein phosphorylation on the assembly of adherens-type junctions. EMBO J. 1992 May;11(5):1733–1742. [PubMed]
  • Weinstein DE, Shelanski ML, Liem RK. Suppression by antisense mRNA demonstrates a requirement for the glial fibrillary acidic protein in the formation of stable astrocytic processes in response to neurons. J Cell Biol. 1991 Mar;112(6):1205–1213. [PMC free article] [PubMed]
  • Westmeyer A, Ruhnau K, Wegner A, Jockusch BM. Antibody mapping of functional domains in vinculin. EMBO J. 1990 Jul;9(7):2071–2078. [PubMed]
  • Wittelsberger SC, Kleene K, Penman S. Progressive loss of shape-responsive metabolic controls in cells with increasingly transformed phenotype. Cell. 1981 Jun;24(3):859–866. [PubMed]
  • Zieske JD, Bukusoglu G, Gipson IK. Enhancement of vinculin synthesis by migrating stratified squamous epithelium. J Cell Biol. 1989 Aug;109(2):571–576. [PMC free article] [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press