Extraction of DNA from formalin-fixed paraffin-embedded tissues and its application to the study of molecular pathology.
| Project/Area Number |
62480139
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Human pathology
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| Research Institution | National Cancer Center Research Institute |
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Principal Investigator |
HIROHASHI Setsuo Pathology Division, National Cancer Center Research Institute, 室長 (70129625)
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| Co-Investigator(Kenkyū-buntansha) |
MUKAI Kiyoshi Pathology Division, National Cancer Center Research Institute (20190837)
YOSHIDA Teruhiko Genetics Division, National Cancer Center Research Institute (10191602)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1988: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1987: ¥3,000,000 (Direct Cost: ¥3,000,000)
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| Keywords | formalin-fixed paraffin-embedded tissue / DNA extraction / oncogene amplification / hepatitis B virus / polymerase chain reaction / 乳癌の予後因子 / がんの悪性度 / Coxの重回帰分析 / 胃癌 / sam / c-erbB-2 / c-myc / 肝癌 / B型肝炎ウイスル / ホルマリン固定パラフィン包理組織 / ドットブロット法 / 癌遺伝子の増幅 / 予後因子 / Nーmyc / cーexbBー2 / hst |
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| Research Abstract |
Quality of DNAs extracted from formalin-fixed paraffin-embedded tissues was examined. The DNAs showd degradation such as fragmentation and were not suitable for southern blot analysis in some cases, but the copy number of amplified genes and presence of foreign viral DNA could be correctly analyzed using dot blot hybridization in all cases (this indicates that the base sequence is preserved even in fragmented DNA). In addition, application of newly devised polymerase chain reaction (PCR) made it possible to amplify short length of DNA to a enormous amount in vitro using DNA extracted formalin-fixed tissues or a single tissue section as a template. Base sequences of the amplified DNA fragments can be easily determined.
Thus, we are able to get the correct informations on genetic abnormalities from formalin-fixed tissues, and it is suggested that diagnoses of diseases will be made by the combination of pathological and genetic analyses of the same material in the future.
The results we have obtained by the application of these methods are as follow. 1) Amplification of N-myc in 51% of neuroblastoma cases correlated with the poor prognosis of patients. 2) Breast cancers carrying amplified c-erbB-2, hst-1 or c-myc showed poor prognosis of patients, and the c-erbB-2 amplification correlated with histological grade of malignancy. 3) Increasing incidence of hepatocellular carcinoma associated with non-A, non-B hepatitis. 4) Amplification of ras gene sequences using polymerase chain reaction and formalin-fixed tissues.
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Report
(3 results)
Research Products
(19 results)
