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Olga Dratviman-Storobinsky, Yoram Cohen, Shahar Frenkel, Efrat Merhavi-Shoham, Shimrit Dadon-Bar El, Natalia Binkovsky, Jacob Pe'er, Nitza Goldenberg-Cohen; The Role of RASSF1A in Uveal Melanoma. Invest. Ophthalmol. Vis. Sci. 2012;53(6):2611-2619. doi: https://doi.org/10.1167/iovs.11-7730.
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© ARVO (1962-2015); The Authors (2016-present)
RASSF1A inactivation in uveal melanoma (UM) is common and methylation-induced. We investigated the effect of RASSF1A re-expression on the UM phenotype in vivo and in vitro.
The phenotypic effect of methylation-induced inactivation of RASSF1A in UM was explored using a stable RASSF1A-expressing UM-15 clone. RASSF1A expression was assessed using QRT-PCR. Proliferation was evaluated in vitro using MTT assays. Additionally, athymic NOD/SCID mice were injected subcutaneously or intraocularly with RASSF1A-expressing and –non-expressing UM-15 clones, and euthanized when tumors reached a volume of 1500 mm3, or at 56 or 46 days, respectively. Tumor tissues, eyes, and livers were analyzed histologically.
In vitro analysis confirmed the lack of RASSF1A expression and full methylation of the RASSF1A promoter region in the UM-15 cell line, which was reversible following treatment with 5-Aza-2-deoxycytidine. Cells expressing exogenous RASSF1A showed slower proliferation than controls and regained sensitivity to cisplatin. Compared to mice injected with control cells, mice treated with UM-15 cells expressing exogenous RASSF1A did not acquire intraocular tumors, and their subcutaneous tumors were relatively delayed and small. Neither group had liver metastases.
UM cells reduced tumorigenicity in the presence of activated RASSF1A. RASSF1A apparently has an important role in the development of UM, and its reactivation might be applied in the development of new treatments.
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