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Camila C. Simoes, J W. Harbour, Ruth Halaban, Antonella Bacchiocchi, Miguel A. Materin; Comparative Study of Oncogenic Mutations and Standard Prognostic Risk Factors for Metastasis in Uveal Melanomas Cell Lines Established by FNAB. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1438.
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(1) To report the establishment of uveal melanoma cell lines by FNAB and (2) to evaluate, in these cell lines, the direct relationship between gene expression profile (GEP) and recognized clinical and histopathologic prognostic factors for metastasis with presence of mutations in genes involved in different uveal melanoma pathways.
From June 2009 to October 2010 patients undergoing FNAB at the time of treatment for primary uveal melanoma had one sample of freshly isolated tissue submitted to cell culture. Cell lines were established and mutational analyses were performed in short-term passages cells. Mutations in the oncogenes B-RAF, N-RAS, CDKN2A, GNA11, GNAQ, PTEN, MITF, TP53, and β-catenin were detect by DNA sequencing. Clinical, histopathologic and GEP signature were reviewed for each patient in whom a new cell line was established.
Five cell lines were established from 5 different patients. At the time of FNAB 2 patients were treated by brachytherapy and 3 by enucleation. Patients ranged in age from 21 to 86 years (mean 51 years).The mean largest basal diameter was 15.5 mm (10 to 20 mm), and mean thickness was 9.1 mm (4.8 to 15 mm). Three tumors were located in the choroid and 2 involved the ciliary body. Three tumors were classified as mixed cell type, and in 2 tumors cytology was inconclusive for acellular material. GEP was categorized as class 1 for 3 tumors, class 2 for 1 tumor and in 1 tumor GEP analysis was not available at the time of treatment. As part of our mutational analysis the first established cell line expressed widely type (WT) BRAF, NRAS, and GNAQ mutation; the second established cell line expressed NRAS, CDKN2A, and WT BRAF and GNAQ mutation. Mutational analysis in these cell lines and in the other ones are still in process. The strongest relationship was between large tumor with class 2 signature and presence of somatic mutation in the genes GNAQ, BRAF and NRAS.
Despite the difficulties, establishment of cell lines by FNAB is a feasible method.Moreover, our study suggest that a combination of unfavorable clinical, histopathologic and GEP with somatic mutations in genes related to metastatic behavior may allow new insights in uveal melanoma metastasis prognostication and could lead to new therapeutic possibilities.
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