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

1997 Fiscal Year Final Research Report Summary

• Project/Area Number: 08670627
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: TOKAI UNIVERSITY
• Principal Investigator: KAGAWA Tatehiro Tokai University School of Medicine Assistant Professor, 医学部, 講師 (30245469)
• Co-Investigator(Kenkyū-buntansha): NISHIZAKI Yasuhiro Tokai University School of Medicine Assistant Professor, 医学部, 講師 (80237693) ; WATANABE Norihito Tokai University School of Medicine Associate Professor, 医学部, 助教授 (90167156)
• Project Period (FY): 1996 – 1997
• Keywords: antisense / K-ras / pancreatic cancer / gene therapy

Research Abstract

1. Establishment and characterization of a pancreatic cancer cell line
A 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 oligodeoxynucleotides
Antisense 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 line
Cells 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.