Ribozyme cleavage leads to decreased expression of fibroblast growth factor receptor 3 in human multiple myeloma cells, which is associated with apoptosis and downregulation of vascular endothelial growth factor

Oligonucleotides. 2005 Spring;15(1):1-11. doi: 10.1089/oli.2005.15.1.

Abstract

The aim of this study was to investigate the fibroblast growth factor receptor 3 (FGFR3) mRNA cleavage by ribozymes targeting FGFR3, effect of growth inhibition and associated with mechanism on multiple myeloma (MM). We designated two ribozyme-expressing plasmids that target the FGFR3 genes, Rz52 and Rz32. In vitro catalytic activity of Rz52 and Rz32 in KMS11 cells decreased FGFR3 mRNA expression to 45% (p < 0.05) and 80% (p < 0.5), respectively, of that of the control. In vivo examination of the Rz52-transfected KMS11 clone showed that FGFR3 mRNA expression decreased to 20% (p < 0.05) of the control. In the Rz52-transfected H929 clone, FGFR3 mRNA decreased to 50% of the control. Protein expression of FGFR3 decreased to 70% of the parental KMS11 and H929 clones. DNA synthesis in the Rz52-transfected KMS11 clone decreased to 20% of that of the control, whereas the viability of cells decreased to 2% (p < 0.01) of that of the control. Ribozyme cleavage-associated increase in apoptosis of Rz52 KMS11 transfectants was twice that of the control. The inhibition of FGFR3 expression by ribozymes was associated with decreased vascular endothelial growth factor (VEGF) expression and upregulation of Flt-1 but not of the KDR receptor. Our data indicate that FGFR3 is an important cell survival and antiapoptotic factor for MM cells and that ribozyme-targeted downregulation of FGFR3 might be useful as a novel therapeutic intervention in MM characterized by t(4;14).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis*
  • Blotting, Western
  • Catalysis
  • Cell Cycle
  • Cell Line
  • Cell Line, Tumor
  • Cell Proliferation
  • Cell Survival
  • Cell-Free System
  • Chromosomes, Human, Pair 14
  • Chromosomes, Human, Pair 4
  • DNA / metabolism
  • Down-Regulation*
  • Humans
  • Immunohistochemistry
  • Models, Genetic
  • Multiple Myeloma / metabolism
  • Multiple Myeloma / pathology*
  • Plasmids / metabolism
  • Polymerase Chain Reaction
  • Protein-Tyrosine Kinases / biosynthesis*
  • Protein-Tyrosine Kinases / genetics*
  • RNA, Catalytic / chemistry*
  • RNA, Messenger / metabolism
  • Receptor, Fibroblast Growth Factor, Type 3
  • Receptors, Fibroblast Growth Factor / biosynthesis*
  • Receptors, Fibroblast Growth Factor / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Thymidine / chemistry
  • Transfection
  • Translocation, Genetic
  • Tumor Cells, Cultured
  • Up-Regulation
  • Vascular Endothelial Growth Factor A / biosynthesis*
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • RNA, Catalytic
  • RNA, Messenger
  • Receptors, Fibroblast Growth Factor
  • Vascular Endothelial Growth Factor A
  • DNA
  • FGFR3 protein, human
  • Protein-Tyrosine Kinases
  • Receptor, Fibroblast Growth Factor, Type 3
  • Thymidine