miR-17 regulates melanoma cell motility by inhibiting the translation of ETV1.
miR-17 regulates melanoma cell motility by inhibiting the translation of ETV1.
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DOI：
10.18632/oncotarget.4147
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发表时间：
2015-08-07
期刊：
影响因子：
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通讯作者：
Markel G
中科院分区：
文献类型：
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作者：
Cohen R;Greenberg E;Nemlich Y;Schachter J;Markel G
黑色素瘤是一种具有高转移潜力的侵袭性恶性肿瘤。 microRNA-17 (miR-17) 是致癌 miR-17/92 簇的成员。在这里，我们研究 miR-17 对黑色素瘤细胞运动的影响。在 WM-266-4 和 624mel 黑色素瘤系中过度表达 miR-17 的成熟或 pri-microRNA 形式可增强细胞运动性，这在伤口愈合和 Transwell 迁移测定中都很明显。 TargetScan 算法预测 PEA3 亚家族成员 ETV1 作为 miR-17 的直接靶标。事实上，将 miR-17 转染到各种黑色素瘤系后，观察到 ETV1 蛋白水平降低了 3-4 倍，而 ETV1 mRNA 表达没有显着变化。双荧光素酶实验证明 miR-17 与 ETV1 3'非翻译区直接结合，通过消除假定结合位点的点突变证实了这一点。这些综合结果表明 miR-17 通过直接翻译抑制来调节 ETV1。此外，在两种黑色素瘤细胞系中，选择性 siRNA 敲低 ETV1 成功地实现了促进细胞迁移的表型复制，而 ETV1 的过度表达则抑制细胞运动和迁移。 ETV1 表达的改变不会影响黑色素瘤的净增殖。总之，我们展示了 miR-17 在黑色素瘤中的新作用，通过靶向 ETV1 蛋白的翻译来促进细胞运动，这可能支持转移的发展。
Melanoma is an aggressive malignancy with a high metastatic potential. microRNA-17 (miR-17) is a member of the oncogenic miR-17/92 cluster. Here we study the effect of miR-17 on melanoma cell motility. Over expression of the mature or pri-microRNA form of miR-17 in WM-266-4 and 624mel melanoma lines enhances cell motility, evident in both wound healing and transwell migration assays. TargetScan algorithm predicts the PEA3-subfamily member ETV1 as a direct target of miR-17. Indeed, a 3–4-fold decrease of ETV1 protein levels are observed following miR-17 transfection into the various melanoma lines, with no significant change in ETV1 mRNA expression. Dual luciferase experiments demonstrate direct binding of miR-17 to the 3′-untranslated region of ETV1, confirmed by abolishing point mutations in the putative binding site. These combined results suggest regulation of ETV1 by miR-17 by a direct translational repression. Further, in both melanoma cell lines ETV1 knockdown by selective siRNA successfully pheno-copies the facilitated cell migration, while overexpression of ETV1 inhibits cell motility and migration. Altered ETV1 expression does not affect melanoma net-proliferation. In conclusion, we show a new role for miR-17 in melanoma, facilitating cell motility, by targeting the translation of ETV1 protein, which may support the development of metastasis.