FAM129B is a novel regulator of Wnt/β-catenin signal transduction in melanoma cells.
FAM129B is a novel regulator of Wnt/β-catenin signal transduction in melanoma cells.
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DOI：
10.12688/f1000research.2-134.v2
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Chien AJ
中科院分区：
文献类型：
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作者：
Conrad W;Major MB;Cleary MA;Ferrer M;Roberts B;Marine S;Chung N;Arthur WT;Moon RT;Berndt JD;Chien AJ
靶向BRAF抑制剂不能在黑色素瘤的临床结果中产生长期持续的改善，这突显了寻找抑制黑色素瘤生长的其他方法的必要性。最近的研究表明，Wnt/β-catenin通路的激活抑制了肿瘤的生长，并与ERK/MAPK通路抑制剂协同促进黑色素瘤细胞的凋亡。因此，Wnt/β-Catenin调节剂的发现可能会促进治疗该病的新方法的发展。为了达到这一目标，我们在人纤维肉瘤细胞中进行了大规模的小干扰核糖核酸(SiRNA)筛选，以寻找β-连环蛋白激活的报告活性的调节因子。结合大规模的小干扰RNA筛选数据，结合磷蛋白质组数据和生物信息学浓缩，确定了一个蛋白质，FAM129B，作为WNT/β-连环蛋白信号的潜在调节者。从功能上，我们证明了在A375和A2058黑色素瘤细胞系中，小干扰RNA介导的FAM129B的敲除抑制了WNT3a介导的β-连环蛋白反应荧光素酶报告基因的激活，并抑制了内源性WNT/β-连环蛋白靶基因AXIN2的表达。我们还证明了FAM129B基因敲除抑制了Wnt3a处理的黑色素瘤细胞的凋亡。这些实验支持FAM129B在将WNT/β-连环蛋白信号与黑色素瘤细胞凋亡联系起来的作用。
The inability of targeted BRAF inhibitors to produce long-lasting improvement in the clinical outcome of melanoma highlights a need to identify additional approaches to inhibit melanoma growth. Recent studies have shown that activation of the Wnt/β-catenin pathway decreases tumor growth and cooperates with ERK/MAPK pathway inhibitors to promote apoptosis in melanoma. Therefore, the identification of Wnt/β-catenin regulators may advance the development of new approaches to treat this disease. In order to move towards this goal we performed a large scale small-interfering RNA (siRNA) screen for regulators of β-catenin activated reporter activity in human HT1080 fibrosarcoma cells. Integrating large scale siRNA screen data with phosphoproteomic data and bioinformatics enrichment identified a protein, FAM129B, as a potential regulator of Wnt/β-catenin signaling. Functionally, we demonstrated that siRNA-mediated knockdown of FAM129B in A375 and A2058 melanoma cell lines inhibits WNT3A-mediated activation of a β-catenin-responsive luciferase reporter and inhibits expression of the endogenous Wnt/β-catenin target gene, AXIN2. We also demonstrate that FAM129B knockdown inhibits apoptosis in melanoma cells treated with WNT3A. These experiments support a role for FAM129B in linking Wnt/β-catenin signaling to apoptosis in melanoma.