目的：探讨鸦胆子油乳剂（BJOE）对食管鳞状细胞癌TE-1细胞增殖、凋亡和自噬的影响及其可能的机制。方法：按干预措施的不同，将TE-1细胞分为对照组、RAPA（自噬激动剂）组、740Y-P（PI3K激活剂）组、BJOE 组、BJOE+RAPA组和BJOE+740Y-P 组。采用FCM、克隆形成、Transwell 实验检测细胞凋亡、增殖、迁移和侵袭能力，qPCR 法检测细胞中PI3K、Akt、mTOR、LC3Ⅰ、LC3Ⅱ、p62、Beclin 1、caspase-3的mRNA表达，WB法检测PI3K、Akt、mTOR及其磷酸化、LC3Ⅱ/Ⅰ、p62、Beclin 1、caspase-3的蛋白表达。结果: 与对照组比较，RAPA组和BJOE 组细胞凋亡率均显著升高（均P<0.01），细胞克隆形成率、迁移和侵袭能力均显著降低（均P<0.01），细胞中PI3K、Akt、mTOR 的mRNA 和蛋白磷酸化水平均显著降低（均P<0.05），p62的mRNA 和蛋白水平显著降低（均P<0.01），LC3Ⅱ/Ⅰ、Beclin 1和caspase-3的mRNA和蛋白水平显著升高（均P<0.05），740Y-P 组的结果则相反（均P<0.05）；与RAPA组或740Y-P 组比较，BJOE+RAPA组或BJOE+740Y-P 组细胞凋亡率显著升高（均P<0.01），克隆形成率、细胞侵袭和迁移能力显著降低（均P<0.01），PI3K、Akt、mTOR的mRNA和蛋白磷酸化水平均显著降低（均P<0.05），p62 的mRNA和蛋白水平均显著降低（均P<0.05），LC3Ⅱ/Ⅰ、Beclin 1 和caspase-3 mRNA和蛋白水平显著升高（均P<0.05）。结论: BJOE显著抑制TE-1细胞增殖、迁移、侵袭并促进细胞凋亡与自噬，其机制可能与抑制PI3K/Akt/mTOR信号通路的激活有关。

Objective: To explore the effects of brucea javanica oil emulsion (BJOE) on proliferation, apoptosis and autophagy of esophageal squamous cell carcinoma TE-1 cells and the possible mechanism. Methods: According to the different interventions TE-1 cells were divided into control group, RAPA (autophagy agonist) group, 740Y-P (PI3K activator) group, BJOE group, BJOE+RAPA group and BJOE+740Y-P group. Cell apoptosis, proliferation, migration and invasion were detected by FCM, clonogenesis and Transwell assay; the mRNA expression of PI3K, Akt, mTOR, LC3Ⅰ, LC3Ⅱ, p62, Beclin 1 and caspase-3 in cells was detected by qPCR; and the protein expression levels of PI3K,Akt, mTOR and their phosphorylation as well as the protein expression of LC3Ⅱ/Ⅰ, p62, Beclin1 and caspase-3 were detected by Western blotting. Results: Compared with the control group, RAPA group and BJOE group exhibited increased cell apoptosis rate (both P<0.01),reduced clone formation rate, cell invasion and migration ability (all P<0.01), decreased mRNA and protein phosphorylation levels of PI3K,Akt, mTOR (all P<0.05) as well as decreased mRNA and protein levels of p62 (all P<0.01), and increased mRNA and protein levels of LC3 Ⅱ/Ⅰ, Beclin 1 and caspase-3 (all P<0.05); however, the results of 740Y-P group were opposite (all P<0.05). Compared with RAPA group or 740Y-P group alone, BJOE+RAPA or BJOE+740Y-P group exhibited increased apoptosis rate (P<0.01), decreased clone formation rate,cell invasion and migration ability (all P<0.01), decreased mRNA and protein phosphorylation levels of PI3K, Akt, mTOR (all P<0.05) as well as decreased mRNA and protein levels of p62 (all P<0.05), and increased mRNA and protein levels of LC3Ⅱ/Ⅰ, Beclin 1 and caspase-3 (all P<0.05). Conclusion: BJOE significantly inhibits the proliferation, invasion, migration and promotes apoptosis and autophagy of TE-1 cells, and the mechanism may be related to inhibition of the activation of PI3K/Akt/mTOR signaling pathway.

国家中医药管理局第七批全国名老中医药专家学术继承项目[No.国中医药人教函（2022）76号]