Synthetic microRNA targeting glioma-associated antigen-1 protein

Methods Mol Biol. 2009:487:435-49. doi: 10.1007/978-1-60327-547-7_21.

Abstract

The transcription factor glioma-associated antigen-1 (Gli-1) mediates activation of the sonic hedgehog (Shh) pathway, a process that precedes the transformation of tissue stem cells into cancerous stem cells and that is involved in early and late epithelial tumorigenesis. Hypothesizing that targeting the 3'-untranslated region (3'-UTR) of Gli-1 mRNA would effectively inhibit epithelial tumor cell proliferation, we evaluated several complementary miRNA molecules for their ability to do so. The synthetic miRNAs and corresponding duplex/small temporal RNAs were introduced as 3-nucleotide (nt) loops into GU-rich portions of the 3'UTR Gli-1 sequence. One particular miRNA (miRNA Gli-1-3548) and its corresponding duplex (Duplex 3548) significantly inhibited proliferation of Gli-1+ ovarian (SK-OV-3) and pancreatic (MiaPaCa-2) tumor cells by delaying cell division and activating late apoptosis in MiaPaCa-2 cells. Here, we describe the design of effective miRNA sequences and their applications as anti-gene agents.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis
  • Cell Line, Tumor
  • Cell Proliferation
  • Female
  • Gene Expression
  • Gene Silencing
  • Genetic Therapy / methods*
  • Humans
  • MicroRNAs / chemical synthesis*
  • MicroRNAs / genetics*
  • Ovarian Neoplasms / genetics*
  • Ovarian Neoplasms / metabolism
  • Pancreatic Neoplasms / genetics*
  • Pancreatic Neoplasms / metabolism
  • RNA, Messenger / genetics
  • Transcription Factors / antagonists & inhibitors*
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Zinc Finger Protein GLI1

Substances

  • GLI1 protein, human
  • MicroRNAs
  • RNA, Messenger
  • Transcription Factors
  • Zinc Finger Protein GLI1