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Detection of 1 mutant cell in normal 10^5〜10^6 cells : Genetic diagnosis using enriched PCR

Research Project

Project/Area Number 10470267
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field Thoracic surgery
Research InstitutionTohoku University

Principal Investigator

SAGAWA Motoyasu  Tohoku University Hospital, Research Associa, 医学部・附属病院, 助手 (70292274)

Co-Investigator(Kenkyū-buntansha) SATO Masami  Tohoku University Hospital, Research Associa, 医学部・附属病院, 助手 (30250830)
藤村 重文  東北大学, 加齢医学研究所, 教授 (40006078)
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 2000: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1999: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1998: ¥2,100,000 (Direct Cost: ¥2,100,000)
Keywordsenriched PCR / PCR / point mutation / enriched PCR / mutation enrichment / ras / DNA ポリメラーゼ / K-ras 遺伝子
Research Abstract

Although sputum cytology and brushing cytology under the bronchoscope are useful for the detection of early lung cancer, it is impossible to detect cancer cells which have mutant DNA, but is not accompanied by the morphological change. On the other hand, it is important to detect very small ratio of mutant cells which exists in many normal cells in the clinical sample. In such case, detection method which has higher sensitivity is required. Enriched polymerase chain reaction method is the technology developed on the basis of the PCR-RFLP method, which can detect 1 mutant cell in normal 105〜106 cells. The principle of this method is as follows. After the samples are amplified (1st PCR), using the primer which introduced restriction enzyme cleavage site, normal DNA (the PCR product) was cut by the restriction enzyme. And then, only abnormal DNA was amplified by the second PCR.The problem of this method is misreading of the Taq DNA polymerase. In our study, DNA polymerase other than Taq was used, because some DNA polymerase has proofreading exonuclease activity. We have investigated various condition for enriched PCR with cultured cell, for the detection of point mutation of K-ras. Then, the investigation was proceeded to the establishment of the condition for the enriched PCR with pathological and cytological specimens.

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] Sagawa M, et al.: "Segmentectomy for roentgenographically occult bronchogenic squamous cell carcinoma"Ann Thorac Surg. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sagawa M, et al.: "CDDP-based chemotherapy/chemoradiotherapy vs.short-term immunochemotherapy in lung cancer"J Cardiovas Surg. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Oyaizu T,Sagawa M, et al.: "Lambert-Eaton syndrome associated with an anterior mediastinal small cell carcinoma"J Thoracic Cardiovas Surg. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Fujimura S,Sagawa M, et al.: "A therapeutic approach to roentgenographically occult squamous cell carcinoma of the lung"Cancer. 89. 2445-2448 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Aikawa H,Sagawa M, et al.: "MDM2 expression is associated with progress of disease and WAF1 expression in resected lung cancer"Int J Mol Med. 5. 631-633 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sato M,Sagawa M, et al.: "Second operation for upper mediastinal lymphadenopathy after complete resection for cancer of the left lung"J Thoracic Cardiovas Surg. 119. 1291-1292 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sagawa M, et al.: "Segmentectomy for roentgenographically occult bronchogenic squamous cell carcinoma."Ann Thorac Surg. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sagawa M, et al.: "A randomized trial of postoperative CDDP-based chemotherapy/chemoradiotherapy vs. short-term immunochemotherapy in lung cancer."J Cardiovasc Surg. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Oyaizu T, Sagawa M, et al.: "Lambert-Eaton syndrome associated with an anterior mediastinal small cell carcinoma."J Thorac Cardiovasc Surg. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Fujimura S, Sagawa M, et al.: "A therapeutic approach to roentgenographically occult squamous cell carcinoma of the lung."Cancer. 89. 2445-2448 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Aikawa H, Sagawa M, et al.: "MDM2 expression is associated with progress of disease and WAF1 expression in resected lung cancer."Int J Mol Med. 5. 631-633 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sato M, Sagawa M, et al.: "Second operation for upper mediastinal lymphadenopathy after complete resection for cancer of the left lung."J Thorac Cardiovasc Surg. 119. 1291-1292 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sagawa M, et al.: "Segmentectomy for roentgenographically occult bronchogenic squamous cell carcinoma"Ann Thorac Surg. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sagawa M, et al.: "CDDP-based chemotherapy/chemoradiotherapy vs.short-term immunochemotherapy in lung cancer"J Cardiovas Surg. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Oyaizu T,Sagawa M, et al.: "Lambert-Eaton syndrome associated with an anterior mediastinal small cell carcinoma"J Thoracic Cardiovas Surg. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Fujimura S,Sagawa M, et al.: "A therapeutic approach to roentgenographically occult squamous cell carcinoma of the lung"Cancer. 89. 2445-2448 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Aikawa H,Sagawa M, et al.: "MDM2 expression is associated with progress of disease and WAF1 expression in resected lung cancer"Int J Mol Med. 5. 631-633 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sato M,Sagawa M, et al.: "Second operation for upper mediastinal lymphadenopathy after complete resection for cancer of the left lung"J Thoracic Cardiovas Surg. 119. 1291-1292 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sagawa M. et al.: "Five-year survivors with resected pN2 nonsmall cell lung carcinoma"Cancer. 85. 864-868 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sato G.,Sagawa M. et al.: "Prognostic value of nucleolar protein p120 in patients wrth・・・・・・"J Clin Oncol. 17. 2721-2727 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Chen Y.,Sagawa M. et al.: "Expression of Bcl-2, Bax, and p53 proteins in carcinogenesis of・・・・・"Anticancer Res. 19. 1351-1356 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Alkawa H.,Sagawa M. et al.: "Immunohistochemical study on tumor angiogenic factors in non・・・・・・"Anticancer Res. 19. 4305-4310 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 佐川元保,藤村重文 他: "呼吸器腫瘍外科学"南江堂. 335 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 佐川元保 他: "気管支鏡による超音波検査"金芳堂. 81 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sagawa,M.et al: "Specific primer design and exonuclease III treatment for・・・・・" Cell Vision. 5・1. 20-24 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sagawa,M.et al: "Sequential loss of heterozygosity in the progression of・・・・" Britishe Journal of Cancer. 78・5. 612-615 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sato,M.et al: "Chromosome bands 3p14.2,9p21,and 13q14 are・・・・" Genes,Chromosomes & Cancer. 23. 367-370 (1998)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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