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

K-ras反义寡聚脱氧核苷酸对胰腺癌细胞株的抗增殖作用

• 批准号: 08670627
• 负责人: KAGAWA Tatehiro
• 金额: $ 1.41万
• 依托单位: TOKAI UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670627/
• 关键词: antisense; K-ras; pancreatic cancer; gene therapy

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.

1.胰腺癌细胞系Pca-S的建立及生物学特性DNA分析显示K-ras密码子12.2由GGT(Gly)突变为GTT(Val)。设计针对K-ras mRNA12密码子的硫代反义寡核苷酸，以针对v-K-ras mRNA12密码子的正义硫代寡核苷酸作为对照。砷对前列腺癌-S细胞系增殖的影响细胞培养在含10%胎牛血清的RPM1-1640中。在第0天和第1天加入不同浓度的AS，在第4天和第7天用四甲基偶氮唑盐比色法检测细胞的增殖情况，在第4天，AS浓度为5和10µM时，细胞增殖抑制达35%，但在第7天，AS的抗增殖作用消失，说明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的抗增殖作用有限。