Fascin phosphorylation sites combine to regulate esophageal squamous cancer cell behavior
Fascin phosphorylation sites combine to regulate esophageal squamous cancer cell behavior
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肌成束蛋白磷酸化位点联合调节食管鳞癌细胞行为
DOI：
10.1007/s00726-017-2398-1
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发表时间：
2017-05-01
期刊：
影响因子：
3.5
通讯作者：
Li, En-Min
中科院分区：
文献类型：
--
作者：
Zeng, Fa-Min;Wang, Xiao-Ning;Li, En-Min
丝状伪足是由F-肌动蛋白捆绑产生的动态膜延伸，并参与癌细胞的迁移、侵袭和转移。肌成束蛋白是丝状伪足中关键的肌动蛋白捆绑蛋白，肌成束蛋白丝氨酸39处的磷酸化被很好地表征为调节肌成束蛋白介导的丝状伪足中的肌动蛋白捆绑。然而，越来越多的证据表明，肌成束蛋白是磷酸化的一些网站。其他位点的磷酸化是否也调节肌成束蛋白功能尚不清楚。在这项研究中，我们发现肌成束蛋白中的四个潜在磷酸化位点，特别是酪氨酸23，丝氨酸38，丝氨酸39和丝氨酸274，调节食管鳞癌细胞的细胞行为和丝状伪足形成。非磷酸化突变的表达在每个四个网站促进锚定独立的生长，细胞运动和丝状伪足的形成，而在这些网站的拟磷酸化突变抑制这些细胞的行为，这意味着肌成束蛋白在食管鳞癌的功能是由肌成束蛋白磷酸化在多个网站。此外，在S38和S39的磷酸化协同调节细胞行为和丝状伪足形成，在S38和S39残基的双重去磷酸化最大限度地增强细胞增殖、迁移和丝状伪足形成，并且在两个磷酸化位点中的任何一个的磷酸化导致增强降低。总之，我们的研究结果表明，肌成束蛋白氨基酸Y23，S38，S39和S274的磷酸化，在组合中，下调食管鳞状细胞癌细胞中的锚定非依赖性生长，细胞迁移和丝状伪足形成的程度。
Filopodia are dynamic membrane extensions generated by F-actin bundling and are involved in cancer cell migration, invasion and metastasis. Fascin is the crucial actin-bundling protein in filopodia, with phosphorylation at fascin serine 39 being well characterized to regulate fascin-mediated actin bundling in filopodia. However, increasing evidence indicates that fascin is phosphorylated at a number of sites. Whether phosphorylation at other sites also regulates fascin function is unknown. In this study, we show that four potential phosphorylation sites in fascin, specifically tyrosine 23, serine 38, serine 39 and serine 274, regulate cell behavior and filopodia formation in esophageal squamous cancer cells. Expression of non-phosphorylatable mutations at each of the four sites promoted anchorage-independent growth, cell motility and filopodia formation, whereas phosphomimetic mutations at each of these sites inhibited these cell behaviors, implying that fascin function in esophageal squamous cancer is regulated by fascin phosphorylation at multiple sites. Furthermore, phosphorylation at S38 and S39 cooperatively regulated cell behavior and filopodia formation, with dual dephosphorylation at both S38 and S39 residues maximally enhancing cell proliferation, migration and filopodia formation, and phosphorylation at any of the two phosphorylatable sites resulting in reduced enhancement. Taken together, our results reveal that phosphorylation at fascin amino acids Y23, S38, S39 and S274, in combination, downregulates the extent of anchorage-independent growth, cell migration and filopodia formation in esophageal squamous cancer cells.