Ezrin Ser66 phosphorylation regulates invasion and metastasis of esophageal squamous cell carcinoma cells by mediating filopodia formation
Ezrin Ser66 phosphorylation regulates invasion and metastasis of esophageal squamous cell carcinoma cells by mediating filopodia formation
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Ezrin Ser66磷酸化通过介导丝状伪足形成调节食管鳞癌细胞的侵袭和转移
DOI：
10.1016/j.biocel.2017.05.018
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发表时间：
2017-07-01
影响因子：
4
通讯作者：
Li, En-Min
中科院分区：
文献类型：
--
作者：
Li, Li-Yan;Xie, Ying-Hua;Li, En-Min
背景Ezrin是连接细胞质膜和肌动蛋白骨架的蛋白，在食管鳞癌的发生、发展中起重要作用。然而，埃兹蛋白S66磷酸化在ESCC肿瘤发生中的作用仍不清楚。方法：通过分析去污剂可溶性/不可溶性组分和胞浆/膜分级分离来检查埃兹蛋白在膜和胞浆组分中的分布。采用免疫荧光和活体成像技术研究ezrin S66磷酸化对细胞丝状伪足中ezrin和肌动蛋白行为的影响。分别采用细胞增殖和迁移实验检测ESCC细胞的增殖、迁移和侵袭能力。结果：Ezrin S66磷酸化增强了Ezrin在ESCC细胞膜上的募集。此外，不可磷酸化的ezrin（S66 A）显著防止丝状伪足形成，以及引起丝状伪足的数量、长度和寿命的减少。此外，功能实验显示，非磷酸化ezrin（S66 A）的表达在体外可显著抑制ESCC细胞的迁移和侵袭，但对ESCC细胞的增殖无明显影响;在体内可显著抑制ESCC细胞的局部侵袭和区域淋巴结转移，但对原发肿瘤的生长无明显影响。Ezrin S66磷酸化促进丝状伪足的形成，有助于调节食管鳞癌细胞的侵袭和转移。
Background. Ezrin, links the plasma membrane to the actin cytoskeleton, and plays an important role in the development and progression of human esophageal squamous cell carcinoma (ESCC). However, the roles of ezrin S66 phosphorylation in tumorigenesis of ESCC remain unclear.Methods: Distribution of ezrin in membrane and cytosol fractions was examined by analysis of detergent soluble/-insoluble fractions and cytosol/membrane fractionation. Both immunofluorescence and live imaging were used to explore the role of ezrin S66 phosphorylation in the behavior of ezrin and actin in cell filopodia. Cell proliferation, migration and invasion of ESCC cells were investigated by proliferation and migration assays, respectively. Tumorigenesis, local invasion and metastasis were assessed in a nude mouse model of regional lymph node metastasis.Results: Ezrin S66 phosphorylation enhanced the recruitment of ezrin to the membrane in ESCC cells. Additionally, non-phosphorylatable ezrin (S66A) significantly prevented filopodia formation, as well as caused a reduction in the number, length and lifetime of filopodia. Moreover, functional experiments revealed that expression of non-phosphorylatable ezrin (S66A) markedly suppressed migration and invasion but not proliferation of ESCC cells in vitro, and attenuated local invasion and regional lymph node metastasis, but not primary tumor growth of ESCC cells in vivo.Conclusion: Ezrin S66 phosphorylation enhances filopodia formation, contributing to the regulation of invasion and metastasis of esophageal squamous cell carcinoma cells.