Department of Oral Implantology, The Affiliated Stomatological Hospital of Kunming Medical University, Kunming, Yunnan 650101, P.R. China.
Department of Stomatology, The Affiliated Yan'an Hospital of Kunming Medical University, Kunming, Yunnan 650051, P.R. China.
Oncol Rep. 2018 Mar;39(3):1356-1368. doi: 10.3892/or.2017.6169. Epub 2017 Dec 20.
The present study aimed to assess the induction of epithelial-mesenchymal transition (EMT), invasion, and metastasis by the chemokine CXCL9/receptor CXCR3 axis in tongue squamous cell carcinoma (TSCC), unveiling the underlying mechanisms and providing new insights into the prevention and treatment of oral cancer metastasis. The expression levels of CXCL9 and CXCR3 in TSCC tissue specimens were determined by immunohistochemistry, assessing differences between samples with cervical lymph node metastasis and those without. Moreover, protein expression or activity in the TSCC Cal-27 cell line was controlled by neutralizing antibodies, gene transfection, or knock-out. Then, alterations of cell proliferation, migration, invasion, and the cytoskeleton were analyzed by CCK-8, cell scratch, Transwell, and cyto-skeleton staining assays, respectively. Alterations of EMT markers (E-cadherin and vimentin) in Cal-27 cells were detected by immunofluorescence and western blotting. In addition, western blotting was utilized to detect protein expression levels of Akt2, p-Akt2, eIF4E and p-eIF4E, and to explore the regulatory roles and mechanisms of the CXCL9/CXCR3 axis in invasion and metastasis. Significantly increased expression levels of CXCL9 and CXCR3 were detected in tissue specimens with lymph node metastasis compared with those without (P<0.01). Overexpression of CXCL9/CXCR3 in Cal-27 cells resulted in cytoskeleton alterations, decreased E-cadherin expression, increased vimentin levels, enhanced migration and invasion (P<0.05), and increased phosphorylated Akt2 and eIF4E levels (P<0.05). These results revealed that in TSCC, the CXCL9/CXCR3 axis could activate the Akt signaling pathway, with EMT and cytoskeleton rearrangement, promoting invasion and metastasis.
本研究旨在评估趋化因子 CXCL9/受体 CXCR3 轴在舌鳞状细胞癌（TSCC）中诱导上皮-间充质转化（EMT）、侵袭和转移的能力，揭示其潜在机制，并为口腔癌转移的预防和治疗提供新的见解。采用免疫组织化学法检测 TSCC 组织标本中 CXCL9 和 CXCR3 的表达水平，评估有颈部淋巴结转移和无颈部淋巴结转移的标本之间的差异。此外，通过中和抗体、基因转染或基因敲除来控制 TSCC Cal-27 细胞系中的蛋白表达或活性。然后，通过 CCK-8、细胞划痕、Transwell 和细胞骨架染色试验分别分析细胞增殖、迁移、侵袭和细胞骨架的变化。通过免疫荧光和 Western blot 检测 Cal-27 细胞中 EMT 标志物（E-钙黏蛋白和波形蛋白）的变化。此外，Western blot 用于检测 Akt2、p-Akt2、eIF4E 和 p-eIF4E 的蛋白表达水平，并探讨 CXCL9/CXCR3 轴在侵袭和转移中的调节作用和机制。与无淋巴结转移的组织标本相比，有淋巴结转移的组织标本中 CXCL9 和 CXCR3 的表达水平显著升高（P<0.01）。Cal-27 细胞中 CXCL9/CXCR3 的过表达导致细胞骨架改变、E-钙黏蛋白表达减少、波形蛋白水平增加、迁移和侵袭增强（P<0.05），以及磷酸化 Akt2 和 eIF4E 水平升高（P<0.05）。这些结果表明，在 TSCC 中，CXCL9/CXCR3 轴可激活 Akt 信号通路，通过 EMT 和细胞骨架重排，促进侵袭和转移。
