2002 Fiscal Year Annual Research Report

遺伝子変異を標的とした大腸癌微小転移の定量的検出法の開発

Research Project

Project/Area Number	14571222
Research Institution	Iwate Medical University
Principal Investigator	樋口太郎岩手医科大学, 医学部, 助手 (40244909)
Co-Investigator(Kenkyū-buntansha)	前沢千早岩手医科大学, 医学部, 講師 (10326647)
Keywords	MGB / ASO / k-ras / p53 / APC
Research Abstract	本研究課題では2年間の研究期間内に大腸癌のリンパ節・門脈血・末梢血中の微小残存病変をP53,k-ras, APC遺伝子の変異を標的にして定量評価可能なASO RQ-PCR(allele-specific oliognucleotide PCR)法を開発することを目的として,平成14年度分として以下の成果を得た. (1)MGB (Minor Groove Binder-conjugated) DNA probeを用いた定量法の開発:conventionalなprobeでは一塩基多型を認識できず,一本鎖DNAを直鎖にし,高いTmを保持できるMGB probeを採用することにした.MGB probeでは20万コピーまでの正常plasmidを認識せず,一方標的変異を含んだplasmidは2 copyまで正確に検出可能であった.K-rasにつていは新しい方法論を開発し,論文を投稿中である. (2)MGB probeの特性:MGB probeは3'端に結合させた場合には不安定であり,5'端に結合させた方がより正確な定量が可能であることが明らかとなった. (3)MGB probeはprimerとしても有用であり,ASO strategy様に開発した新規primerを作製中である. MGBテクノロジーを用いた微小残存病変の検出法は,従来型の検出法に比較して鋭敏で優れた方法である.次年度以降はp53,APC遺伝子での解析を行い,本法の有用性を臨床検体で検討する.

Research Products
(6 results)

All Other

All Publications (6 results)

[Publications] Wada K, Maesawa C, Satoh T, Akasaka T, Masuda T.: "A case of primary cutaneous CD30+ T-cell lymphoproliferative disorder with features of granulomatous slack skin disease"Br J Dermatol. 147. 998-1002 (2002)
[Publications] Tarusawa M, Yashima A, Endo M, Maesawa C.: "Quantitative assessment of minimal residual disease in childhood lympholid malignancies using on allele-specific oligonucleotide real-time quantitative polymerase chain reaction"Int J Hematol. 75. 166-173 (2002)
[Publications] Sugai T, Uesugi N, Nakamura S, Habano W, Jiao YF, Higuchi, T, et al.: "Evolution of DNA ploidy state and DNA index in colorectal adenomas and carcinomas using the crypt isolation technique : New hypothesis in colorectal tumorigenesis"Pathol Int. 53. 154-162 (2003)
[Publications] Nakatani K, Hagihara S, Sando S, Yamaguchi K, Maesawa C, Saito I: "Induction of a remarkable conformational change in a human telomeric sequence by the binding of naphthyridine dimer : inhibition on the elongation of a telomeric repeat by telomerase"J Am Chem Soc. 125. 662-666 (2003)
[Publications] Maesawa C, Inaba T, Sato H, Iijima S, Ishida K, Terashima M, Sato R, et al.: "A rapid biosensor chip assay for measuring of telomerase activity using surface plasmon resonance"Nucleic Acids Res. 31. E4-4 (2003)
[Publications] Ishikawa K, Koyama T, Masuda T.: "Prevalence of HBV genotypes in asymptomatic carrier residents and their clinical characteristics during long-term follow-up : the revelence to change in the HBeAg/anti-HBe system"Hepatol Res. 24. 1 (2002)

2002 Fiscal Year Annual Research Report

遺伝子変異を標的とした大腸癌微小転移の定量的検出法の開発

Principal Investigator

樋口 太郎 岩手医科大学, 医学部, 助手 (40244909)

Research Products

[Publications] Wada K, Maesawa C, Satoh T, Akasaka T, Masuda T.: "A case of primary cutaneous CD30+ T-cell lymphoproliferative disorder with features of granulomatous slack skin disease"Br J Dermatol. 147. 998-1002 (2002)

[Publications] Tarusawa M, Yashima A, Endo M, Maesawa C.: "Quantitative assessment of minimal residual disease in childhood lympholid malignancies using on allele-specific oligonucleotide real-time quantitative polymerase chain reaction"Int J Hematol. 75. 166-173 (2002)

[Publications] Sugai T, Uesugi N, Nakamura S, Habano W, Jiao YF, Higuchi, T, et al.: "Evolution of DNA ploidy state and DNA index in colorectal adenomas and carcinomas using the crypt isolation technique : New hypothesis in colorectal tumorigenesis"Pathol Int. 53. 154-162 (2003)

[Publications] Nakatani K, Hagihara S, Sando S, Yamaguchi K, Maesawa C, Saito I: "Induction of a remarkable conformational change in a human telomeric sequence by the binding of naphthyridine dimer : inhibition on the elongation of a telomeric repeat by telomerase"J Am Chem Soc. 125. 662-666 (2003)

[Publications] Maesawa C, Inaba T, Sato H, Iijima S, Ishida K, Terashima M, Sato R, et al.: "A rapid biosensor chip assay for measuring of telomerase activity using surface plasmon resonance"Nucleic Acids Res. 31. E4-4 (2003)

[Publications] Ishikawa K, Koyama T, Masuda T.: "Prevalence of HBV genotypes in asymptomatic carrier residents and their clinical characteristics during long-term follow-up : the revelence to change in the HBeAg/anti-HBe system"Hepatol Res. 24. 1 (2002)

樋口太郎岩手医科大学, 医学部, 助手 (40244909)