The antisense c-Ha-ras oligodeoxynucleotides inhibit the proliferation of the gastrocarcinoma-DNA-transformed rat 3-3 cells and reduce the p21 expression of the cells

Sci China B. 1991 Jan;34(1):35-41.

Abstract

The Rat 3-3 is a secondary transformant of the rat fibroblast cell line (Rat-1) transfected with total DNA of a gastrocarcinoma cell line BGC-823. The cells over-express the c-Ha-ras oncogene which contains point mutation at the 12th codon. In order to determine how the activated c-Ha-ras oncogene expression governs the cell's transformation, two pentadecadeoxy-nucleotides AS-1 and AS-2 were synthesized. AS-1 was complementary to the single strand of the first three codons and the upstream sequence close to the ribosome binding site of c-Ha-ras mRNA. AS-2 was complementary to the 3' end of the first intron and the 5' end of the second exon of c-Ha-ras unripe RNA enclosed in the nucleus. The oligonucleotides could block either the translation of c-Ha-ras mRNA or the splicing of c-Ha-ras unripe RNA, thus inhibiting the expression of the activated c-Ha-ras oncogene and the proliferation of the transformed cells Rat 3-3. The inhibitory effect increased with a growing concentration of the antisense oligodeoxynucleotide (from 4-10 mumol/L) and reached its peak at 12 h after Rat 3-3 was treated with AS-1 and AS-2. This effect became weaker afterwards. The p21 level, product of c-Ha-ras, in the Rat 3-3, was examined by ELISA method. The result shows that the amount of p21 in the growth-inhibited cells is about 30% of that of the control cells.

MeSH terms

  • Animals
  • Cell Division / drug effects
  • DNA, Neoplasm / biosynthesis
  • Dose-Response Relationship, Drug
  • Fibroblasts / cytology
  • Gene Expression Regulation, Neoplastic / drug effects
  • Genes, ras*
  • Oligonucleotides, Antisense / pharmacology*
  • Protein Biosynthesis / drug effects
  • Proto-Oncogene Proteins p21(ras) / analysis*
  • RNA Splicing / drug effects
  • Rats
  • Stomach Neoplasms / genetics
  • Stomach Neoplasms / pathology*
  • Transfection
  • Tumor Cells, Cultured / drug effects

Substances

  • DNA, Neoplasm
  • Oligonucleotides, Antisense
  • Proto-Oncogene Proteins p21(ras)