1989 Fiscal Year Final Research Report Summary
In situ DNA cytofluorometry in paraffin sections and its application to malignacy diagnosis.
| Project/Area Number |
62440030
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Human pathology
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
FUJITA Setsuya Kyoto Prefectural University of Medicine Department of Pathology, 医学部, 教授 (00079716)
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| Project Period (FY) |
1987 – 1989
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| Keywords | DNA contents / cytofluorometry / in situ measurement / diagnosis of cancer / microtomography / confocal microscopy / laser scanning |
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| Research Abstract |
It has recently been made clear that by measuring DNA contents of cells one can differentiate malignant changes from benign ones. Benign changes including non-neoplasmic and neoplasmic alteration of cells in the gastro-intestinal epithelium never show aneuploidy nor polyploidy. Extrapolating this result, we can infer that measurements of DNA contents of cells in suspicious changes such as dysplasia or incipient cancers showing no overt atypia in histology may reveal their true nature whether benign or malignant.
To carry out this kind of investigation on paraffin sections which are common and important goods among pathologists, there have been serious obstacles. DNA cytofluorometry is straight-forward inapplicable to in situ measurement of DNA in paraffin sections, due to overlapping of nuclei, non-specific light-loss occurring in the sections.
This year, as a continuation of the former year project, we applied confocal microscopy to get microtomography of fluoresently labelled paraffin
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sections to calculate total intensity of all the pixel values that are above certain threshold and estimated DNA content of a cell. Or by applying normal cytofluorometry to metaphase figures in a section in order to ascertain validity of the confocal measurement.
As a result, it is confirmed that by this technique in situ DNA measurements are possible and that this technique actually revealed precise distribution of malignant cell clones (better to say subclones). Application of this technique has made it clear that human cancers start as an apparently benign, quasi-diploid tumor, which grows rather slowly, to progress gradually toward more malignant tumors, by mutating their DNA from quasi-diploid to neuploid conditions. The investigation is proved very important to the understanding of behaviors of the human cancer in vivo.
Together with these results of the research, the present investigation suggests that further improvement of the confocal microscope and dyes that bind to DNA would be important to correct diagnosis of otherwise ambiguous tumor-identification and to proper understanding of the entire course of human cancer progression in the human body. Less
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