Intracellular expression of the T-cell factor-1 RNA aptamer as an intramer

Mol Cancer Ther. 2006 Sep;5(9):2428-34. doi: 10.1158/1535-7163.MCT-05-0204.

Abstract

T-cell factor (TCF)-1 protein forms the transcriptional complex with beta-catenin and regulates the expression of diverse target genes during early development and carcinogenesis. We have selected previously an RNA aptamer that binds to the DNA-binding domain of TCF-1 and have shown that it interfered with binding of TCF-1 to its specific DNA recognition sequences in vitro. As an approach to modulate the transcription by TCF/beta-catenin complex in the cells, we have developed the RNA expression vector for stable expression of RNA aptamer inside of the mammalian cells. High level of RNA was expressed as an intramer in the fusion with the stable RNA transcript. The RNA intramer inhibited TCF/beta-catenin transcription activity as shown by luciferase assay. It also modulated the expression of TCF/beta-catenin target genes, such as cyclin D1 and matrix metalloproteinase-7, as predicted to be as an effective inhibitor of the TCF function. In addition, it efficiently reduced the growth rate and tumorigenic potential of HCT116 colon cancer cells. Such RNA intramer could lead to valuable gene therapeutics for TCF/beta-catenin-mediated carcinogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Nucleotide / biosynthesis*
  • Aptamers, Nucleotide / genetics
  • Base Sequence
  • Cell Growth Processes / genetics
  • Gene Expression
  • HCT116 Cells
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • T Cell Transcription Factor 1 / biosynthesis
  • T Cell Transcription Factor 1 / genetics*
  • Transcription, Genetic
  • beta Catenin / genetics
  • beta Catenin / metabolism

Substances

  • Aptamers, Nucleotide
  • T Cell Transcription Factor 1
  • beta Catenin