| Project/Area Number |
12557101
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
General surgery
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
MAEHARA Yoshihiko Kyushu University, Surgery and Science, professor, 大学院・医学研究院, 教授 (80165662)
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| Co-Investigator(Kenkyū-buntansha) |
TOKUNAGA Eriko Kyushu University, Hospital, Instructor, 医学部附属病院, 助手 (50325453)
KAKEJI Yoshihiro Kyushu University, Hospital, Assistant professor, 医学部附属病院, 講師 (80284488)
北村 薫 九州大学, 医学部・附属病院, 講師 (70234276)
高橋 郁雄 九州大学, 医学部・附属病院, 講師 (70325439)
池田 陽一 九州大学, 医学部・附属病院, 助手 (50311840)
古賀 聡 九州大学, 医学部・附属病院, 医員
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥13,300,000 (Direct Cost: ¥13,300,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2000: ¥8,200,000 (Direct Cost: ¥8,200,000)
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| Keywords | Real-time PCR / micrometastasis / peripheral blood / bone marrow / microsatellite instability / mismatch repair / genetic analysis / 複製エラー / DNAミスマッチ修復 / 大腸癌 / HNPCC / 遺伝子ノックアウトマウス / micrometastasis / 定量的RT-PCR / 微小癌細胞 |
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| Research Abstract |
Micrometastasis; Real time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect disseminated cancer cells in bone marrow and peripheral blood. We compared CEA, CK18, CK19 and CK20 expression using 20 of cell lines in order to select the most appropriate. In an analysis of CK 18 mRNA expression in peripheral venous blood in 13 healthy volunteers, we found that CK 18 mRNA was much less expressed than in cancer cell lines. However, the expression in all samples was at a level which was also detected using conventional RT-PCR. It would thus seem that not only qualitative, but also quantitative analysis, of the target mRNA is important to detect micrometastasis. Quantitative RT-PCR methods will make comparisons of the possible differences in expression levels of the target gene. For clinical applications, much further study is needed.
Microsalellite Instability; Using high-resolution fluorescent microsatellite analysis (HFRMA), we studied over 800 patients with sporadic breast, gastric, colon, lung and liver cancer. In this system, several devices were prepared to improve reproducibility of polymerase chain reaction products, migration accuracy of electrophoresis, and characteristics of the detection system. All the microsatellite changes observed in this system can be classified into two types: type A with relatively small changes in microsatellite sequences observed in limited loci and type B with drastic and widely dispersed changes. The former was thought to be connected to abnormal activity in DNA mismatch repair (MMR).
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