[摘 要] 目的：探讨川楝素（TSN）对食管鳞状细胞癌（ESCC）KYSE150细胞增殖、凋亡、迁移和侵袭的影响及其分子机制。 方法：通过CCK-8法、克隆形成和EdU实验检测不同浓度TSN（0.062 5、0.125、0.25 μmol/L）对KYSE150细胞增殖的影响，流式细 胞术、划痕实验和Transwell实验检测TSN对KYSE150细胞凋亡、迁移和侵袭的影响。通过GEPIA数据库数据分析食管癌组织 中低氧诱导因子1α（HIF1A）的表达，qPCR法检测人食管上皮细胞Het-1A和KYSE150细胞，以及TSN处理的各组KYSE150细胞 中HIF1A mRNA的表达水平。WB法检测TSN对HIF1A的上游信号通路AKT/mTOR和下游与细胞迁移、侵袭和凋亡相关蛋白 表达的影响。结果：经不同浓度TSN处理后，KYSE150细胞的增殖、迁移和侵袭能力均显著降低（P < 0.05或P < 0.01），细胞凋 亡率均显著升高（P < 0.05或P < 0.01）。HIF1A mRNA在KYSE150细胞中呈高表达（P < 0.05），TSN处理后KYSE150细胞中 HIF1A mRNA表达显著降低（P < 0.05或P < 0.01）。TSN能够显著抑制KYSE150细胞中HIF1A及其上游通路关键蛋白p-AKT、 p-mTOR及下游迁移、侵袭和凋亡相关蛋白N-cadherin、vimentin、Bcl-2、caspase-3 的表达均显著下调，E-cadherin表达上调 （P < 0.05或P < 0.01）。结论：TSN通过AKT/mTOR信号通路下调HIF1A表达，从而抑制KYSE150细胞增殖、迁移、侵袭并诱导 细胞凋亡。

[Abstract] Objective: To investigate the effects of toosendanin (TSN) on the proliferation, apoptosis, migration and invasion of esophageal squamous cell carcinoma (ESCC) KYSE150 cells, and to elucidate its underlying molecular mechanisms. Methods: CCK-8 assay, colony formation assay, and EdU assay were used to assess the effects of varying TSN concentrations (0.062 5, 0.125, and 0.25 μmol/L) on KYSE150 cell proliferation. The impacts of TSN on the apoptosis, migration, and invasion of KYSE150 cells were evaluated using flow cytometry, wound healing assay, and Transwell chamber assay, respectively. The expression of hypoxia-inducible factor 1 alpha (HIF1A) in esophageal cancer tissues was analyzed using the GEPIA database. qPCR was used to detect the expression level of HIF1A mRNA in human esophageal epithelial Het-1A and KYSE150 cells, and in TSN-treated KYSE150 cells. Western blot (WB) was performed to detect the effects of TSN on the upstream signaling pathway AKT/mTOR of HIF1A and the expression of downstream proteins related to cell migration, invasion, and apoptosis. Results: TSN of varying concentrations significantly inhibited proliferation, migration, and invasion of KYSE150 cells and promoted apoptosis in a dose-dependent manner (P < 0.05 or P < 0.01). HIF1A mRNA was highly expressed in KYSE150 cells, and its expression was significantly downregulated after TSN treatment (P < 0.05 or P < 0.01). TSN markedly downregulated the expression of HIF1A and key upstream signaling proteins p-AKT and p-mTOR. In addition, TSN significantly suppressed the expression of downstream proteins associated with cell migration, invasion, and apoptosis, including N-cadherin, vimentin, Bcl-2, and caspase-3, while upregulating the expression of E-cadherin (P < 0.05 or P < 0.01). Conclusion: TSN inhibits the proliferation, migration, and invasion, and induces apoptosis in ESCC KYSE150 cells by down-regulating HIF1A expression through suppression of the AKT/mTOR signaling pathway.

[基金项目] 南充市2023年市级科技研发计划专项（No. 23JCYJPT0026）