3'-End conjugates of minimally phosphorothioate-protected oligonucleotides with 1-O-hexadecylglycerol: synthesis and anti-ras activity in radiation-resistant cells

Bioconjug Chem. 2000 Mar-Apr;11(2):153-60. doi: 10.1021/bc990106n.

Abstract

Activation of the ras oncogene has been implicated in many types of human tumors. It has been shown that downmodulation of ras expression can lead to the reversion of the transformed phenotype of these tumor cells. Antisense oligodeoxyribonucleotides (ODNs) can inhibit gene expression by hybridization to complementary mRNA sequences. To minimize toxicity associated with all-phosphorothioated ODNs and improve cellular uptake, we used partially phosphorothioate (PPS)-modified ODNs having an additional hydrophobic tail at the 3'-end (PPS-C(16)). The PPS ODNs are protected against degradation by PS internucleotide linkages at both the 3'- and 5'-ends and additionally stabilized at internal pyrimidine sites, which are the major sites of endonuclease cleavage. Here we show that anti-ras PPS-C(16) ODN retains the high sequence-specificity of PPS ODNs and provides maximal inhibition of Ras p21 synthesis with minimal toxicity even without the use of a cellular uptake enhancer. Moreover, treatment of T24, a radiation-resistant human tumor cell line that carries a mutant ras gene, with anti-ras PPS-C(16) ODN resulted in a reduction in the radiation resistance of the cells in vitro. We also demonstrate that the growth of RS504 (a human c-Ha-ras transformed NIH/3T3 cell line) mouse tumors was significantly inhibited by the combination of intratumoral injection of anti-ras PPS-C(16) ODN and radiation treatment. These findings indicate the potential of this combination of antisense and conventional radiation therapy as a highly effective cancer treatment modality.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions / chemistry*
  • Animals
  • Codon, Initiator / drug effects
  • Codon, Initiator / metabolism
  • Female
  • Genes, ras / genetics
  • Glyceryl Ethers / chemistry*
  • Humans
  • Mice
  • Mice, Nude
  • Molecular Structure
  • Neoplasms, Experimental / drug therapy
  • Neoplasms, Experimental / genetics
  • Oligoribonucleotides / chemistry
  • Oligoribonucleotides / pharmacology*
  • Oligoribonucleotides, Antisense / metabolism
  • Oligoribonucleotides, Antisense / pharmacology
  • Oncogene Protein p21(ras) / antagonists & inhibitors
  • Oncogene Protein p21(ras) / biosynthesis
  • Phenotype
  • RNA, Messenger / metabolism
  • Radiation Tolerance / drug effects
  • Structure-Activity Relationship
  • Thionucleotides / chemical synthesis*
  • Thionucleotides / pharmacology*
  • Transfection
  • Tumor Cells, Cultured / drug effects
  • Tumor Cells, Cultured / radiation effects

Substances

  • 3' Untranslated Regions
  • Codon, Initiator
  • Glyceryl Ethers
  • Oligoribonucleotides
  • Oligoribonucleotides, Antisense
  • RNA, Messenger
  • Thionucleotides
  • Oncogene Protein p21(ras)
  • chimyl alcohol