The role of RASSF1A in uveal melanoma.
The role of RASSF1A in uveal melanoma.
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DOI：
10.1167/iovs.11-7730
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发表时间：
2012-10-03
影响因子：
4.4
通讯作者：
Goldenberg-Cohen, Nitza
中科院分区：
文献类型：
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作者：
Dratviman-Storobinsky, Olga;Cohen, Yoram;Goldenberg-Cohen, Nitza
目的：RASSF1A在葡萄膜黑色素瘤（UM）中失活是常见的，并且是甲基化诱导的。我们研究了在体内和体外的UM表型的RASSF1A再表达的效果。方法：甲基化诱导的失活RASSF1A在UM的表型效应进行了探讨使用一个稳定的RASSF1A表达UM-15克隆。使用QRT-PCR评估RASSF1A表达。使用MTT测定法在体外评价增殖。此外，无胸腺NOD/SCID小鼠皮下或眼内注射表达RASSF1A和不表达UM-15克隆，并在肿瘤体积达到1500 mm3时，或分别在56或46天时实施安乐死。肿瘤组织，眼睛和肝脏进行了组织学分析。结果：在体外分析证实，缺乏RASSF1A的表达和RASSF1A启动子区域的UM-15细胞系，这是可逆的治疗后，5-氮杂-2-脱氧胞苷的甲基化。表达外源性RASSF1A的细胞表现出比对照更慢的增殖，并恢复了对顺铂的敏感性。与注射对照细胞的小鼠相比，用表达外源性RASSF1A的UM-15细胞处理的小鼠没有获得眼内肿瘤，并且它们的皮下肿瘤相对延迟且较小。两组均无肝转移。结论：UM细胞在激活RASSF1A的存在下降低致瘤性。RASSF1A在UM的发生发展中起重要作用，其重新激活可能用于开发新的治疗方法。
PURPOSE: 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.METHODS: 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 mm(3), or at 56 or 46 days, respectively. Tumor tissues, eyes, and livers were analyzed histologically.RESULTS: 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.CONCLUSIONS: 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.