Preferences help
enabled [disable] Abstract
Number of results
2006 | 53 | 2 | 311-316
Article title

Regulation of β1 integrin expression in endothelial cells by chimeric tRNAVal ribozyme

Title variants
Languages of publication
To downregulate expression of the β1 integrin subunit in endothelial cells, plasmid encoding the ribozyme cassette containing hammerhead ribozyme flanked at the 5' terminus by tRNAVal and at the 3' terminus by constitutive transport element sequences was constructed. When used to transfect immortalized human endothelial cell line EA.hy 926, it selectively blocked the synthesis of the β1 integrin subunit and thus inhibited migration and proliferation of the cells. Thus, this construct may be a valuable tool to control the proangiogenic phenotype of stimulated endothelial cells.

Physical description
  • Department of Molecular and Medical Biophysics, Medical University of Lodz, Lodz, Poland
  • Department of Bioorganic Chemistry, Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Lodz, Poland
  • Bayless KJ, Salazar R, Davis GE (2000) RGD-dependent vacuolation and lumen formation observed during endothelial cell morphogenesis in three-dimensional fibrin matrices involves the αvβ3 and α5β1 integrins. Am J Pathol 156: 1673-1683.
  • Bloch W, Forsberg E, Lentini S, Brakebusch C, Martin K, Krell HW, Weidle UH, Addicks K, Fassler R (1997) β1 integrin is essential for teratoma growth and angiogenesis. J Cell Biol 139: 265-278.
  • Bouck N, Stellmach V, Hsu SC (1996) How tumors become angiogenic. Adv Cancer Res 69: 135-174.
  • Cieslak M, Niewiarowska J, Nawrot M, Koziolkiewicz M, Stec WJ, Cierniewski CS (2002) DNAzymes to β1 and β3 mRNA downregulate expression of the targeted integrins and inhibit cell capillary tube formation in fibrin and Matrigel J Biol Chem 277: 6779-6787.
  • Cieślak M, Szymanski J, Adamiak R, Cierniewski CS (2003) Structural rearrangements of the “10-23” DNAzyme to β3 integrin subunit mRNA induced by cations and their relations to the catalytic activity. J Biol Chem 278: 47987-47996.
  • Dallabrida SM, DeSouza MA, Farrell DH (2000) Expression of antisense to integrin subunit β3 inhibits microvascular endothelial cell capillary tube formation in fibrin. J Biol Chem 275: 32281-32288.
  • Eliceiri BP, Cheresh DA (1999) The role of αv integrins during angiogenesis: insights into potential mechanisms of action and clinical development J Clin Invest 103: 1227-1212.
  • Hanahan D, Folkman J (1996) Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell 86: 353-364.
  • Huttenlocher A, Ginsberg MH, Horwitz AF (1996) Modulation of cell migration by integrin-mediated cytoskeletal linkages and ligand-binding affinity. J Cell Biol 134: 1551-1562.
  • Hynes RO (1992) Integrins: versatility, modulation, and signaling in cell adhesion. Cell 69: 11-25.
  • Kato Y, Kuwabara T, Warashina M, Toda H, Taira K (2001) Relationships between the activities in vitro and in vivo of various kinds of ribozyme and their intracellular localization in mammalian cells. J Biol Chem 276: 15378-15385.
  • Koivisto L, Heino J, Hakkinen L, Larjava H (1994) The size of the intracellular beta 1-integrin precursor pool regulates maturation of beta 1-integrin subunit and associated alpha-subunits. Biochem J 300: 771-779.
  • Koseki S, Tanabe T, Tani K, Asano S, Shioda T, Nagai Y, Shimada T, Ohkawa J, Taira K (1999) Factors governing the activity in vivo of ribozymes transcribed by RNA polymerase III. J Virol 73: 1868-1877.
  • Kuwabara T, Warashina M, Koseki S, Sano M, Ohkawa J, Nakayama K, Taira K (2001) Significantly higher activity of a cytoplasmic hammerhead ribozyme than a corresponding nuclear counterpart: engineered tRNAs with an extended 3’ end can be exported efficiently and specifically to the cytoplasm in mmamalian cells. Nucleic Acids Res 29: 2780-2788.
  • Lauffenburger DA, Horwitz AF (1996) Cell migration: a physically integrated molecular process. Cell 84: 359-369.
  • Leong KG, Hu X, Li L, Noseda M, Larrivée B, Hull C, Hood L, Wong F, Karsan A (2002) Activated Notch4 inhibits angiogenesis: role of β1-integrin activation. Mol Cell Biol 22: 2830-2841.
  • Missol-Kolka E, Sochanik A, Szala S, Szary J (2003) In Gene Therapy (Szala S, eds) Wydawnictwo Naukowe PWN SA, Warsaw (in Polish).
  • Nawrot B, Antoszczyk S, Maszewska M, Kuwabara T, Warashina M, Taira K, Stec WJ (2003) Efficient inhibition of β-secretase gene expression in HEK293 cells by tRNAVal-driven and CTE-helicase associated hammerhead ribozymes. Eur J Biochem 270: 3962-3970.
  • O’Brien ER, Garvin MR, Dev R, Stewart DK, Hinohara T, Simpson JB, Schwartz SM (1994) Angiogenesis in human coronary atherosclerotic plaques. Am J Pathol 145: 883-894.
  • Palecek SP, Huttenlocher A, Horwitz AF, Lauffenburger DA (1998) Physical and biochemical regulation of integrin release during rear detachment of migrating cells. J Cell Sci 111: 929-940.
  • Panda D, Kundu GC, Lee BI, Peri A, Fohl D, Chackalaparamil I, Mukherjee BB, Li XD, Mukherjee DC, Seides S, Rosenberg J, Stark K, Mukherjee AB (1997) Potential roles of osteopontin and alphaVbeta3 integrin in the development of coronary artery restenosis after angioplasty. Proc Natl Acad Sci USA 94: 9308-9313.
  • Rola M, Kuzmak J (2001) The application of the hammerhead ribozymes as antivirial tools. Postepy Biochem 47: 282-291 (in Polish).
  • Saelman EU, Keely PJ, Santoro SA (1995) Loss of MDCK cell alpha 2 beta 1 integrin expression results in reduced cyst formation, failure of hepatocyte growth factor/scatter factor-induced branching morphogenesis, and increased apoptosis. J Cell Sci 108: 3531-3540.
  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning, A Laboratory Manual, 2nd edn, Cold Spring Harbor Laboratory Press.
  • Shimayama T, Nishikawa S, Taira K (1995) Generality of the NUX rule: kinetic analysis of the results of systematic mutations in the trinucleotide at the cleavage site of hammerhead ribozymes. Biochemistry 34: 3649-3654.
  • Warashina M, Kuwabara T, Kato Y, Sano M, Taira K (2001) RNA-protein-hybrid ribozymes that efficiently cleave any mRNA independently of the structure of the target RNA. Proc Natl Acad Sci USA 98: 5572-5577.
  • Zuker M, Mathews D, Turner D (1999) Algorithms and thermodynamics for RNA secondary structure prediction: a practical guide. In RNA Biochemistry and Biotechnology (Barciszewski J, Clarc B, eds) NATO Asi Series, Kulwer Academic Publishers.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.