Department of Otolaryngology-Head and Neck Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, P.R. China.
Exp Biol Med (Maywood). 2021 Jun;246(11):1244-1252. doi: 10.1177/1535370221992703. Epub 2021 Feb 16.
Laryngeal squamous cell cancer (LSCC) is a common carcinoma with high morbidity and mortality. Metastasis constitutes the major cause of death and poor prognosis among patients with LSCC. Recent evidence confirms critical function of Wnt1-inducible signaling protein 1 (WISP1) in several cancers. However, its contribution in LSCC metastasis remains unclear. Specimens of tumor tissues and adjacent normal mucosa were collected from patients with LSCC. The mRNA and protein levels were determined using quantitative real-time PCR and Western blot, respectively. RNA interference was applied to silence the expression of WISP1 and TGF-β, and recombinant adenovirus was used to overexpress WISP1 in human LSCC cell line TU212 cells. Cell invasion and migration were determined by transwell assay. High expression of WISP1 was observed in LSCC tissues, especially in those from metastatic groups. Ectopic expression of WISP1 enhanced invasion and migration of TU212 cells. On the contrary, WISP1 knockdown reduced numbers of invasive and migrated cells. Additionally, elevation of WISP1 depressed the expression of epithelial marker E-cadherin and increased levels of mesenchymal marker vimentin in TU212 cells, whereas WISP suppression yielded the opposite effects. Further analysis corroborated that WISP1 overexpression enhanced activation of TGF-β-Smad signaling by increasing expression of TGF-β1, p-Smad2, and p-Smad3, which was abrogated following WISP1 down-regulation. Moreover, TGF-β1 exposure facilitated LSCC cell invasion and migration. Notably, blockage of the TGF-β-Smad pathway by si-TGF-β overturned WISP-1-evoked epithelial-to-mesenchymal transition (EMT), and subsequent cell invasion and migration. These findings highlight the pro-metastatic function of WISP1 in LSCC by regulating cell invasion and migration via TGF-β-Smad-mediated EMT, supporting a promising invention target for LSCC therapy.
喉鳞状细胞癌（LSCC）是一种常见的癌，发病率和死亡率都很高。转移是 LSCC 患者死亡和预后不良的主要原因。最近的证据证实 Wnt1 诱导信号蛋白 1（WISP1）在几种癌症中具有重要功能。然而，它在 LSCC 转移中的作用尚不清楚。收集 LSCC 患者的肿瘤组织和相邻正常黏膜标本。分别采用实时定量 PCR 和 Western blot 测定 mRNA 和蛋白水平。应用 RNA 干扰沉默 WISP1 和 TGF-β 的表达，并用重组腺病毒过表达人 LSCC 细胞系 TU212 细胞中的 WISP1。通过 Transwell 测定细胞侵袭和迁移。在 LSCC 组织中观察到 WISP1 的高表达，特别是在转移组中。异位表达 WISP1 增强了 TU212 细胞的侵袭和迁移能力。相反，WISP1 敲低减少了侵袭和迁移细胞的数量。此外，WISP1 的升高降低了 TU212 细胞中上皮标志物 E-钙粘蛋白的表达，增加了间充质标志物波形蛋白的水平，而 WISP 抑制则产生了相反的效果。进一步分析证实，WISP1 通过增加 TGF-β1、p-Smad2 和 p-Smad3 的表达，增强 TGF-β-Smad 信号的激活，而下调 WISP1 则消除了这种作用。此外，TGF-β1 暴露促进了 LSCC 细胞的侵袭和迁移。值得注意的是，si-TGF-β 阻断 TGF-β-Smad 通路逆转了 WISP1 诱导的上皮间质转化（EMT）以及随后的细胞侵袭和迁移。这些发现强调了 WISP1 通过 TGF-β-Smad 介导的 EMT 调节细胞侵袭和迁移在 LSCC 中的促转移功能，为 LSCC 治疗提供了有前途的靶点。
