| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Research Abstract |
It has been reported that betam actin which was cloned from murine melanoma cell line B16F1 with low metastatic ability suppressed metastatisis to the lung, the invasiveness of collagen gels, in vivo invasiveness in murine system. In order to investivate the effect of betam actin on human melanoma cells, we transfected the betam actin cDNA with SV40 promotor into human melanoma cell lines. First, we chose one cell line named 28.1 with high transfection efficacy, and examined in vitro cell growth, and tumor growth in nude mice. Then, we co-transfected betam actin cDNA (10-20mug) and pSV_2 neo (1mug) into 28.1 cell line by calcium phosphate precipitation method, and isolated neomycine-resistant clones. These cloned were examined by anti-betam actin antibody whether betam actin is expressed in these cells or not. However, no clones expressed betam actin. These results suggested that betam actin may suppress the growth of human melanoma cells, although there are several possibilities. In order confirm this sugestion, we should constract betam cDNA with an inducer in future.
We also focused on alpha-smooth muscle actin in human melanoma and observed that alpha-smooth muscle actin was decreased in human malignant melanoma. Then, we analyzed human melanomas in various stages by immunostaing, and found that in primary and metastatic melanoma, the vessel walls were negative for alpha-smooth muscle actin, while in melanoma in situ the vessel walls were positive for alpha-smooth muscle actin. These findings suggest that the analysis of alpha-smooth muscle actin is useful for the determination of the stages of human malignant melanomas.
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