先前的研究支持信号蛋白 3A (SEMA3A) 在包括口腔鳞状细胞癌 (OSCC) 在内的多种肿瘤中具有肿瘤抑制作用。然而，SEMA3A 在 OSCC 中的作用及其潜在分子机制的深入表征尚缺乏。使用定量实时PCR、蛋白质印迹测定和免疫组织化学检测基因和蛋白质表达。使用 Transwell 评估 OSCC 细胞转移，并使用管形成测定法测定人脐静脉内皮细胞 (HUVEC) 的血管生成。使用生物信息学分析预测分子之间的相互作用，并使用荧光素酶活性实验和RNA免疫沉淀测定进行验证。在构建小鼠肺转移肿瘤模型后，使用苏木精和伊红染色评估肺转移。 OSCC 细胞中 SEMA3A 表达降低，其过度表达导致 OSCC 细胞上皮间质转化（EMT）、迁移和侵袭以及 HUVEC 血管生成受到抑制。 miR-32-5p被鉴定为SEMA3A的上游分子，长非编码RNA NR2F2反义RNA 1（NR2F2-AS1）被验证为miR-32-5p的上游基因。进一步的实验表明，NR2F2-AS1过表达对EMT、OSCC细胞迁移、侵袭、HUVEC血管生成以及小鼠肿瘤生长和转移的抑制作用是通过miR-32-5p/SEMA3A轴介导的。总之，NR2F2-AS1 可能会通过 miR-32-5p/SEMA3A 轴减弱小鼠 OSCC 细胞转移和 HUVEC 血管生成，并抑制肿瘤生长和转移。© 2024 作者。约翰·威利出版的《高雄医学科学杂志》

Previous studies have supported a tumor-suppressive role of semaphorin 3A (SEMA3A) in several tumors including oral squamous cell carcinoma (OSCC). However, in-depth characterization of the role of SEMA3A in OSCC and the underlying molecular mechanisms is lacking. Gene and protein expressions were detected using quantitative real-time PCR, western blot assay, and immunohistochemistry. OSCC cell metastasis was evaluated using Transwell and angiogenesis of human umbilical vein endothelial cells (HUVECs) was determined using tube formation assay. The interactions among molecules were predicted using bioinformatics analysis and validated using luciferase activity experiment and RNA immunoprecipitation assay. Pulmonary metastasis was evaluated using hematoxylin and eosin staining after constructing a lung metastasis tumor model in mice. SEMA3A expression was decreased in OSCC cells and its overexpression led to suppression of epithelial-mesenchymal transition (EMT), migration, and invasion of OSCC cells and angiogenesis of HUVECs. miR-32-5p was identified as an upstream molecule of SEMA3A and long non-coding RNA NR2F2 antisense RNA 1 (NR2F2-AS1) was validated as an upstream gene of miR-32-5p. Further experiments revealed that the inhibitory effects of NR2F2-AS1 overexpression on EMT, migration, invasion of OSCC cells, and angiogenesis of HUVECs as well as tumor growth and metastasis in mice were mediated via the miR-32-5p/SEMA3A axis. To conclude, NR2F2-AS1 may attenuate OSCC cell metastasis and angiogenesis of HUVECs and suppress tumor growth and metastasis in mice via the miR-32-5p/SEMA3A axis.© 2024 The Author(s). The Kaohsiung Journal of Medical Sciences published by John Wiley & Sons Australia, Ltd on behalf of Kaohsiung Medical University.