miR-375 Regulates Invasion-Related Proteins Vimentin and L-Plastin
miR-375 Regulates Invasion-Related Proteins Vimentin and L-Plastin
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DOI：
10.1016/j.ajpath.2017.02.019
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发表时间：
2017-07-01
影响因子：
6
通讯作者：
Segall, Jeffrey E.
中科院分区：
文献类型：
--
作者：
Jimenez, Lizandra;Lim, Jihyeon;Segall, Jeffrey E.
侵袭是晚期头颈部鳞状细胞癌（HNSCC）的标志。我们先前确定，相对较低的miR-375表达与患者预后不良相关。具有增加的miR-375表达的HNSCC细胞具有较低的侵袭特性和受损的侵袭柄活性。我们使用稳定同位素标记细胞培养物中的氨基酸和反相液相色谱质谱法，评估了miR-375表达对UM-SCC-1细胞中蛋白质水平的影响。miR-375表达的增加与参与细胞组装和组织、死亡和存活以及运动的蛋白质的下调相关。两种侵袭相关蛋白，波形蛋白和L-plastin，被miR-375强烈下调。荧光素酶报告基因分析表明，高miR-375表达降低了波形蛋白启动子活性，表明波形蛋白是miR-375的间接靶点。Runt相关转录因子1（RUNX 1）是一个潜在的miR-375直接靶点，其敲低可降低波形蛋白和L-plastin的表达。癌症基因组图谱HNSCC数据库中的数据显示，miR-375表达与RUNX 1、波形蛋白和L-plastin RNA表达之间存在显著负相关。这些临床相关性验证了我们的体外模型发现，并支持miR-375抑制RUNX 1水平，导致波形蛋白和L-plastin表达减少的机制。此外，RUNX 1，L-plastin和vimentin的敲低导致体外细胞侵袭显著减少，表明miR-375调节参与HNSCC侵袭的特定蛋白质的功能意义。
Invasion is a hallmark of advanced head and neck squamous cell carcinoma (HNSCC). We previously determined that low relative miR-375 expression was associated with poor patient prognosis. HNSCC cells with increased miR-375 expression have lower invasive properties and impaired invadopodium activity. Using stable isotope labeling with amino acids in cell culture and reverse-phase liquid chromatography mass spectrometry, we assessed the impact of miR-375 expression on protein levels in UM-SCC-1 cells. Increased miR-375 expression was associated with down-regulation of proteins involved in cellular assembly and organization, death and survival, and movement. Two invasion-associated proteins, vimentin and L-plastin, were strongly down-regulated by miR-375. Luciferase reporter assays demonstrated that high miR-375 expression reduced vimentin promoter activity, suggesting that vimentin is an indirect target of miR-375. Runt-related transcription factor 1 (RUNX1) is a potential miR-375 direct target, and its knockdown reduced vimentin and L-plastin expression. Data in The Cancer Genome Atlas HNSCC database showed a significant inverse correlation between miR-375 expression and RUNX1, vimentin, and L-plastin RNA expression. These clinical correlations validate our in vitro model findings and support a mechanism in which miR-375 suppresses RUNX1- levels, resulting in reduced vimentin and L-plastin expression. Furthermore, knockdown of RUNX1, L-plastin, and vimentin resulted in significant reductions in cell invasion in vitro, indicating the functional significance of miR-375 regulation of specific proteins involved in HNSCC invasion.