| Budget Amount *help |
¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1992: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1991: ¥500,000 (Direct Cost: ¥500,000)
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| Research Abstract |
We studied cell kinetics in human skin cancer using the following three methods, (1) in vitro bromodeoxyuridine labeling, (2) DNA quantification by flow cytometry, and (3) silver staining of nucleolar organizer region (AgNOR).
The DNA synthesizing cells in benign and malignant keratotic diseases including psoriasis, seborrheic keratosis, Bowen's disease and basal cell carcinoma as well as normal skin were studied to understand the pathophysiology of these diseases. Cells in S-phase were labelled by incubating small pieces of excised skin tumors in the tissue culture medium containing 100 muM 5-bromo-2'-deoxyuridine (BrdUrd) under 3 atmospheric pressures of 95%O_2-5%CO_2 for one hour at 37ﾟC. Incorporated BrdUrd was demonstrated with an immunohistochemical method using anti-BrdUrd monoclonal antibody.
We analyzed the DNA labelling using the labelling index and labelling pattern. The labelling pattern was examined to determine if the polarity in labelling was preserved or lost. Although ma
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lignant diseases tended to show higher DNA labelling index, neither DNA labelling index nor the loss of polarity alone was associated with the malignancy. However, high DNA labelling index concomitant with the loss of polarity in labelling in the tumor seemed to be characteristic of the malignancy.
We studied the presence of DNA-aneuploidy using flow cytometry mainly in squamous cell carcinoma and malignant melanoma. Both diseases tended to show DNA-aneuploidy in advanced diseases. Intra-epidermal carcinomas including Bowen's disease did not show DNA-aneuploidy with flow cytometry, however it does not necessarily mean the absence of DNA-aneuploidy in these diseases. It could be possible that diploid pattern due to surrounding normal cells covered the DNA-aneuploidy due to small number of malignant cells.
We applied AgNOR, which stained acidic protein associated with rRNA transcription, to malignant melanomas and basal cell carcinomas. In melanoma and basal cell carcinoma, the number of AgNOR was significantly higher than melanocytic nevi and normal epidermis respectively. Less
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