Queen Mary University of London, Barts and The London School of Medicine and Dentistry, Centre for Clinical and Diagnostic Oral Sciences, Institute of Dentistry, London, UK.
Department of Pharmaceutical and Biological Chemistry, UCL School of Pharmacy, London, UK.
Oncogene. 2014 May 1;33(18):2363-74. doi: 10.1038/onc.2013.186. Epub 2013 Jun 10.
Desmoglein 3 (Dsg3), the pemphigus vulgaris antigen, has recently been shown to be upregulated in squamous cell carcinoma (SCC) and has been identified as a good tumor-specific marker for clinical staging of cervical sentinel lymph nodes in head and neck SCC. However, little is known about its biological function in cancer. The actin-binding protein Ezrin and the activator protein 1 (AP-1) transcription factor are implicated in cancer progression and metastasis. Here, we report that Dsg3 regulates the activity of c-Jun/AP-1 as well as protein kinase C (PKC)-mediated phosphorylation of Ezrin-Thr567, which contributes to the accelerated motility of cancer cells. Ectopic expression of Dsg3 in cancer cell lines caused enhanced phosphorylation at Ezrin-Thr567 with concomitant augmented membrane protrusions, cell spreading and invasive phenotype. We showed that Dsg3 formed a complex with Ezrin at the plasma membrane that was required for its proper function of interacting with F-actin and CD44 as Dsg3 knockdown impaired these associations. The increased Ezrin phosphorylation in Dsg3-overexpressing cells could be abrogated substantially by various pharmacological inhibitors for Ser/Thr kinases, including PKC and Rho kinase that are known to activate Ezrin. Furthermore, a marked increase in c-Jun S63 phosphorylation, among others, was found in Dsg3-overexpressing cells and the activation of c-Jun/AP-1 was further supported by a luciferase reporter assay. Taken together, our study identifies a novel Dsg3-mediated c-Jun/AP-1 regulatory mechanism and PKC-dependent Ezrin phosphorylation that could be responsible for Dsg3-associated cancer metastasis.
桥粒芯糖蛋白 3(Dsg3)是寻常性天疱疮的抗原,最近研究表明其在鳞状细胞癌(SCC)中上调,并被鉴定为头颈部 SCC 颈哨淋巴结临床分期的良好肿瘤特异性标志物。然而,其在癌症中的生物学功能知之甚少。细胞骨架结合蛋白 Ezrin 和激活蛋白 1(AP-1)转录因子与癌症进展和转移有关。在这里,我们报告 Dsg3 调节 c-Jun/AP-1 的活性以及蛋白激酶 C(PKC)介导的 Ezrin-Thr567 的磷酸化,这有助于癌细胞的快速运动。在癌细胞系中异位表达 Dsg3 导致 Ezrin-Thr567 的磷酸化增强,同时伴随着膜突起、细胞铺展和侵袭表型的增强。我们表明 Dsg3 在质膜上与 Ezrin 形成复合物,这是其与 F-肌动蛋白和 CD44 相互作用的适当功能所必需的,因为 Dsg3 敲低会损害这些关联。PKC 和 Rho 激酶等已知激活 Ezrin 的丝氨酸/苏氨酸激酶的各种药理学抑制剂可显著抑制 Dsg3 过表达细胞中 Ezrin 磷酸化的增加。此外,在 Dsg3 过表达细胞中发现 c-Jun S63 磷酸化等显著增加,c-Jun/AP-1 的激活进一步得到荧光素酶报告基因检测的支持。总之,我们的研究确定了一种新的 Dsg3 介导的 c-Jun/AP-1 调节机制和 PKC 依赖性 Ezrin 磷酸化,这可能是 Dsg3 相关癌症转移的原因。