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J.–C.A. Marshall, A. Nantel, P.L. Blanco, J. Ash, S.R. Cruess, E. Achenson, M.N. Burnier, Jr; Transcriptional Profiling of the Evolution of Uveal Melanoma From Cell Lines to Intraocular Tumors to Metastasis . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3390.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Uveal melanoma cells must undergo several genetic changes before they are able to produce metastasis. The aim of this study is to measure the transcriptional profiles of the human cell line 92.1 as it evolved from growth in vitro, to the development of intraocular primary tumors, followed by lung metastatic tumors in a rabbit model of uveal melanoma. Methods: Human 19k microarrays and universal human reference RNA were used to measure the diversity between four cell lines, derived from uveal melanomas (92.1, MKT–BR, OCM–2, SP6.5) and a fifth, originally established from normal melanocytes (UW–1). 92.1 was then used to produce primary intraocular tumors and metastasis in immunosuppressed rabbits. Five samples from both the intraocular and metastatic tumors were then subjected to transcriptional profiling analysis and the results were compared to those of the cultured cell lines. Results: Statistical analysis demonstrated that all five cell lines had virtually the same expression profiles, suggesting that UW–1 has undergone transformations that have made it virtually indistinguishable from the uveal melanoma cell lines. Clustering of individual sample profiles by Principal Components Analysis clearly shows a gradual evolution in the abundance of specific transcripts as the cells adapt from one environment to the next. A very stringent ANOVA cut off (p value < 0.001) was used to identify 555 transcripts that best distinguish one stage from another. The changes from the cell lines to primary tumors are extensive. The abundance of many melanocyte markers, including CD63, gradually decreases as cells evolve which might be indicative of dedifferentiation. Other interesting tumor progression markers include the small GTPase RAB26 and the NF–kB modulator IL1RAP as well as known metastatic markers such as Kangai 1, a tumor suppressor, the adaptor protein IRS–2 and the NME1 nucleoside diphosphate kinase. Conclusions: The UW–1 cell line, used as a normal melanocyte, has undergone transformational changes, which explains its ability to develop tumors in our experimental model. In addition we have produced a detailed analysis of the molecular changes that take place as human uveal melanoma cells evolve from a primary tumor to metastasis including the decrease in expression of specific melanoma markers. Future research will focus on elaborating the role of these markers in the establishment of metastasis.
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