Application of DNA microarray technique to mutagenesis assessment.

• Project/Area Number: 14570305
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Hygiene
• Research Institution: KEIO UNIVERSITY
• Principal Investigator: NAKASHIMA Hiroshi Keio University, School of Medicine, Instructor, 医学部, 助手 (80217710)
• Co-Investigator(Kenkyū-buntansha): OMAE Kazuyuki Keio University, School of Medicine, Professor, 医学部, 教授 (60118924)
• Project Period (FY): 2002 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥4,100,000 (Direct Cost: ¥4,100,000); Fiscal Year 2003: ¥2,000,000 (Direct Cost: ¥2,000,000); Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000)
• Keywords: genotoxicity / DNA chip / expression profile / n-methyl-nitrosourea / mtomycin C / bleomycin / UVC / UVA / 変異原

Research Abstract

High or low concentration mitomycin C (MMC) was exposed to lymphoblast stimulated by 4.0 micro g/ml of phytohemagglutinin (PHA). Total RNA was recovered 24 after exposure and expression profile was analyzed with DNA microarray. Prothymosin alpha showed significant change. Same experiment was done with 2.5 micro g/ml of PHA, because no probe relating cell cycle arrest or DNA repair was identified. However, significant change was observed only for coproporphyrinogen oxidase and 28S rRNA. MMC causes inter strand cross-link for double strand DNA but its distortion is said to be small. This might be the reason for the negative result.
High or low concentration n-methyl-nitrosourea (MNU) was exposed to lymphoblast stimulated by 2.5 micro g/ml of PHA. Total RNA was recovered 24 after exposure and expression profile was analyzed with DNA microarray. MNU is famous with O6 methylation of guanine residue and oxidative stress is induced by MNU. Probes related with cell cycle arrest, DNA repair or oxidative stress response showed significant change.
Reanalysis of BLM, UVA or UVC data revealed corresponding cellular response to the lesion.