Anti-proliferative effect of K-ras antisense oligodeoxynucleotides on pancreatic cancer cell line

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1. Establishment and characterization of a pancreatic cancer cell lineA pancreatic cancer cell line, Pca-S was established from a 60 year-old woman bearing pancreatic carcinoma. The DNA analysis revealed a mutation from GGT (Gly) to GTT (Val) in K-ras codon 12.2. Phosphorothioate oligodeoxynucleotidesAntisense phosphorothioate oligodeoxynucleotides (AS) were designed against codon 12 of K-ras mRNA.Sense phosphorothioate oligodeoxynucleotides against codon 12 of v-K-ras mRNA were used as the controls.3. Effect of AS on proliferation of Pca-S cell lineCells were cultured in RPMl-1640 containing 10% fetal calf serum. AS were added on days 0 and 1 at various concentrations, and the proliferation of cells was evaluated using MTT on days 4 and 7. AS with concentrations of 5 and 10 muM specifically inhibited the proliferation up to 35% on day 4. However, on day 7, the anti-proliferative effects of AS were abolished, indicating that the effect of AS was not lasting. In summary, antisense phosphorothioate oligodeoxynucleotides against v-K-ras codon 12 would have anti-proliferative effect on pancreatic cancer cells with limited duration.