1988 Fiscal Year Final Research Report Summary
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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| Keywords | formalin-fixed paraffin-embedded tissue / DNA extraction / oncogene amplification / hepatitis B virus / polymerase chain reaction |
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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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