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| 431 The 67 Annual Meeting of Japanese Association for Laboratory Animal Science P50 Nanog + melanoma cell lines as a standard model for quantitative evaluation of metastatic potential ○ Mikako Saito,Tomohiro Hatakenaka,Nahoko Matsuki,Seiya Minagawa Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology Various stresses are thought to enhance malignancy of cancer cells with high metastatic potential. However, it is difficult to evaluate the effects of various factors on the metastatic potential in vivo. Here we focus the use of Nanog overexpressing mouse melanoma cell lines, NanogF1, NanogF10, NanogBL6. In vitro tests revealed that the metastatic potential was enhanced by Nanog overexpression. Differential gene expression between NanogBL6 and BL6 was analyzed by transcriptome sequencing. Among the highest 31 up/down-regulated genes, TGF-β1 was only one that could directly generate an excretable cytokine TGF-β1. TGF-β1 expression was confirmed by qPCR analysis and its involvement in the direct regulation of metastatic potential was analyzed by developing a TGF-β1 knockdown cell line (shTGF-β1F10). A relationship between the TGF-β1 expression and the number of metastatic colonies suggested a multi-role of TGF-β1 in the regulation of metastatic potential in vivo. In summary, the functional properties of Nanog overexpressing melanoma cells were clarified comprehensively. The range of metastatic potential change could be defined as 0 to 200 [metastatic colony number/organ]. This strongly suggests the feasibility of the present cell lines as an experimental standard for the quantitative evaluation of various factors with suppressive or enhancing effects on the metastatic potential in vivo.