[摘 要] 目的：探讨月见草素B调控磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）/核因子-κB（NF-κB）通路对卵巢癌SKOV3细胞恶性生物学行为的影响。方法：CCK-8法测定月见草素B对SKOV3细胞的半数抑制浓度（IC50），参考IC??值及相关文献确定后续实验剂量。SKOV3细胞分为对照组，月见草素B低（6.25 μmol/L）、中（12.5 μmol/L）、高（25 μmol/L）剂量组，PI3K抑制剂 （LY294002）组，月见草素B高剂量 + PI3K激活剂（740Y-P）组。CCK-8法、5-乙炔基-2'-脱氧尿苷（EdU）染色法检测各组细胞增殖能力；Transwell侵袭实验、划痕实验、流式细胞术分别检测各组细胞侵袭、迁移能力与凋亡情况；RT-qPCR检测各组细胞中增殖相关基因细胞周期蛋白D1（Cyclin D1）、迁移相关基因迁移侵袭增强子1（MIEN1）及凋亡相关基因p53 mRNA表达情况；Western blotting检测各组细胞中PI3K/AKT/NF-κB通路相关蛋白p-PI3K、p-AKT、NF-κB p65的表达。结果：月见草素B处理SKOV3细胞24 h的IC??值为28.70 μmol/L。与对照组相比，月见草素B低、中、高剂量组及LY294002组SKOV3细胞增殖能力、侵袭细胞数及划痕愈合率均降低（均P < 0.05），Cyclin D1、MIEN1 mRNA及p-PI3K、p-AKT、NF-κB p65蛋白表达下调（均P < 0.05），而细胞凋亡率与p53 mRNA表达则升高（均P < 0.05）；与LY294002组相比，高剂量月见草素B对SKOV3细胞增殖能力、侵袭细胞数及划痕愈合率的抑制作用更显著（均P < 0.05），对Cyclin D1、MIEN1 mRNA及p-PI3K、p-AKT、NF-κB p65蛋白的下调及对细胞凋亡率与p53 mRNA表达的上调均更明显（均P < 0.05）。与月见草素B高剂量组相比，月见草素B高剂量+740Y-P组SKOV3细胞增殖能力、侵袭细胞数及划痕愈合率升高（均P < 0.05），Cyclin D1、MIEN1 mRNA及p-PI3K、p-AKT、NF-κB p65蛋白表达上调（均P < 0.05），细胞凋亡率与p53 mRNA表达降低（P < 0.05）。结论：月见草素B可能通过抑制PI3K/AKT/NF-κB通路，抑制卵巢癌SKOV3细胞增殖、侵袭与迁移，并诱导细胞凋亡。

[Abstract] Objective: To investigate the effect of oenothein B on the malignant biological behavior of ovarian cancer SKOV3 cells by regulating the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/nuclear factor-kappa B (NF-κB) pathway. Methods: The half-maximal inhibitory concentration (IC50) of oenothein B on SKOV3 cells was determined by the CCK-8 assay, and the subsequent experimental doses were determined based on the IC50 value and relevant literature. SKOV3 cells were divided into the control group, low-dose oenothein B group (6.25 μmol/L), medium-dose oenothein B group (12.5 μmol/L), high-dose oenothein B group (25 μmol/L), PI3K inhibitor (LY294002) group, and high-dose oenothein B combined with PI3K activator (740Y-P) group. The cell proliferation ability of each group was detected by CCK-8 assay and 5-ethynyl-2'-deoxyuridine (EdU) staining. Transwell invasion assay, wound healing assay and flow cytometry were used to detect cell invasion, migration ability and apoptosis in each group, respectively. RT- qPCR was applied to detect the mRNA expression levels of proliferation-related gene Cyclin D1, migration-related gene migration and invasion enhancer 1 (MIEN1), and apoptosis-related gene p53 in each group. Western blotting was used to determine the expression levels of PI3K/AKT/NF-κB pathway-related proteins including p-PI3K, p-AKT and NF-κB p65 in each group. Results: The IC50 value · · 670 [[PAGE_INDEX=2 FILE=37dae296-6188-4ec2-9b39-45b08e05f9a4.pdf]] 张颖, 等. 月见草素B调控PI3K/AKT/NF-κB通路对卵巢癌SKOV3细胞恶性生物学行为的影响of oenothein B against SKOV3 cells after 24 h of treatment was 28.70 μmol/L. Compared with the control group, the proliferation ability, number of invasive cells and wound healing rate of SKOV3 cells were significantly decreased in the low-, medium- and high- dose oenothein B groups as well as the LY294002 group (all P < 0.05); the mRNA expression levels of Cyclin D1 and MIEN1, as well as the protein expression levels of p-PI3K, p-AKT and NF-κB p65 were markedly downregulated (all P < 0.05), while the cell apoptosis rate and p53 mRNA expression were remarkably upregulated (all P < 0.05). Compared with the LY294002 group, the high-dose oenothein B exhibited more pronounced inhibitory effects on cell proliferation, invasion, and migration (all P < 0.05), as well as greater downregulation of Cyclin D1 and MIEN1 mRNA and p-PI3K, p-AKT, and NF- κB p65 protein expression, and more significant upregulation of apoptosis rate and p53 mRNA expression (all P < 0.05). Compared with the high-dose oenothein B group, the high-dose oenothein B + 740Y-P group showed increased cell proliferation, invasion, and migration (all P < 0.05), upregulated Cyclin D1 and MIEN1 mRNA and p-PI3K, p-AKT, and NF-κB p65 protein expression (all P < 0.05), and decreased apoptosis rate and p53 mRNA expression (all P < 0.05). Conclusion: Oenothein B may inhibit the PI3K/AKT/NF- κB signaling pathway, thereby suppressing the proliferation, invasion and migration of ovarian cancer SKOV3 cells and inducing cell apoptosis.

[基金项目] 青海省卫生健康科研项目（2023-wjzdx-17）