目的：探讨叉头框K1（FOXK1）在喉鳞状细胞癌（LSCC）中的表达、临床意义及对细胞增殖、迁移、侵袭的影响，并阐明其分子机制。方法：RT-qPCR、免疫组化检测LSCC组织及LSCC细胞TU177中FOXK1表达，分析其与临床病理参数及预后的相关性；通过瞬时转染构建FOXK1过表达与敲低细胞模型，通过MTS法、克隆形成实验、划痕愈合实验和Transwell实验检测细胞恶性生物学行为的变化；采用RT-qPCR检测上皮-间充质转化（EMT）相关因子mRNA的表达；采用WB法检测PI3K/AKT信号通路相关蛋白的表达；结合生物信息学预测与功能回复实验，验证FOXK1对AKT2的调控作用。结果：FOXK1在LSCC组织及细胞中高表达（P < 0.01），且主要定位于细胞核；FOXK1高表达与较晚TNM分期相关（P < 0.05），且FOXK1高表达患者5年生存率较低（P < 0.05）；过表达FOXK1显著增强TU177细胞增殖、迁移与侵袭能力并诱导EMT（P < 0.01或P < 0.05），敲低FOXK1则产生相反的效应；FOXK1可上调AKT2表达并激活PI3K/AKT信号通路；敲低AKT2可部分逆转FOXK1过表达对细胞增殖、迁移、侵袭及EMT 进程的促进作用（P < 0.01 或P < 0.05）。结论：FOXK1 在LSCC 中高表达，与肿瘤进展及不良预后密切相关。FOXK1可能通过上调AKT2激活PI3K/AKT信号通路，诱导EMT，从而促进LSCC细胞的恶性生物学行为。

Objective: To investigate the effects of FOXK1 on the proliferation, migration and invasion of laryngeal squamous cell carcinoma (LSCC) and the prognosis of patients, as well as its molecular mechanism. Methods: RT-qPCR and immunohistochemistry were applied to determine FOXK1 expression in LSCC tissues and TU177 cells, with subsequent analysis of its association with clinicopathological parameters and patient prognosis. FOXK1 overexpression and knockdown vectors were constructed and transfected into LSCC cell line TU177. MTS assay, colony formation assay, scratch wound healing assay, and Transwell assay were employed to detect changes in the malignant biological behaviors of cells. RT-qPCR was employed to detect the mRNA expression of epithelial- mesenchymal transition (EMT)-related factors, and Western blotting was used to detect the protein expression of PI3K/AKT signaling pathway-related proteins. Combined with bioinformatics prediction and functional rescue experiments, the regulatory effect of FOXK1 on AKT2 was verified. Results: FOXK1 was highly expressed in LSCC tissues and cells (P < 0.01), and was mainly localized in the cell nucleus; high FOXK1 expression was associated with a higher TNM stage (P < 0.05), and a lower 5-year survival rate (P < 0.05); FOXK1 overexpression significantly promoted the proliferation, migration and invasion of TU177 cells and induced EMT (P < 0.01 or P < 0.05), whereas FOXK1 knockdown produced the opposite effects; FOXK1 upregulated AKT2 expression and activated the PI3K/ AKT signaling pathway; knockdown of AKT2 partially reversed the promoting effects of FOXK1 overexpression on proliferation, migration, invasion and EMT progression (P < 0.01 or P < 0.05). Conclusion: FOXK1 is highly expressed in LSCC and is closely associated with tumor progression and poor prognosis. FOXK1 may activate the PI3K/AKT pathway by upregulating AKT2 expression, thereby inducing EMT and promoting the malignant biological behaviors of LSCC cells.

河北省医学科学研究课题计划（20250114）