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

1.胰腺癌细胞系的建立和鉴定胰腺癌细胞系Pca-S是从60岁患有胰腺癌的女性建立的。DNA分析显示K-ras基因第12.2位密码子由GGT（Gly）突变为GTT（瓦尔）。硫代磷酸寡核苷酸针对K-ras mRNA第12位密码子设计反义硫代磷酸寡核苷酸（AS），以v-K-ras mRNA第12位密码子的正义硫代磷酸寡核苷酸作为对照. AS对Pca-S细胞系增殖的影响细胞在含10%胎牛血清的RPM 1 -1640中培养。在第0天和第1天加入不同浓度的AS，并在第4天和第7天使用MTT评价细胞增殖。浓度为5和10 μ M的AS在第4天特异性抑制增殖达35%。然而，在第7天，AS的抗增殖作用被取消，表明AS的作用不持久。综上所述，针对v-K-ras密码子12的反义硫代寡核苷酸对胰腺癌细胞具有有限的抗增殖作用。

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.