目的 基于体内外模型，探讨蒲公英甾醇对非小细胞肺癌的作用机制及其与Wnt/β-catenin信号传导的相关性。方法 用不同浓度（0、5、10、20、40、80 μmol/L）的蒲公英甾醇处理A549细胞，优化药物浓度选择。将A549细胞分为对照组、蒲公英甾醇（10、20、40 μmol/L）、蒲公英甾醇（40 μmol/L）＋CHIR99021（10 μmol/L）组。通过平板克隆实验检测增殖能力，运用流式细胞技术分析凋亡情况，结合划痕实验和Transwell小室法分别测定迁移和侵袭活性。利用Western blotting测定Wnt/β-catenin信号转导途径关键分子的表达量。通过构建非小细胞肺癌荷瘤小鼠模型，进一步评估蒲公英甾醇的体内抑瘤效果，并证实Wnt/β-catenin通路在其中起关键介导作用。结果 蒲公英甾醇在5～80 μmol/L浓度下对A549细胞活性具有显著抑制作用（P＜0.05），IC₅₀值为33.95 μmol/L。相较于对照组，蒲公英甾醇（10、20、40 μmol/L）组检测克隆形成数、划痕愈合率、侵袭细胞数以及Wnt通路关键分子（β-catenin、Cyclin D1、c-Myc）蛋白表达水平及GSK-3β磷酸化程度均显著降低，而细胞凋亡率显著上升（P＜0.05）。CHIR99021能够逆转蒲公英甾醇对A549细胞的抑制效应。在动物实验中，与模型组相比，蒲公英甾醇组小鼠的皮下移植瘤体积、瘤质量显著降低（P＜0.05）；HE染色结果显示，蒲公英甾醇组的炎症浸润程度显著减轻；β-catenin、c-Myc、Cyclin D1蛋白表达量及GSK-3β磷酸化均显著下降（P＜0.05）。结论 蒲公英甾醇通过调控Wnt/β-catenin信号转导途径，发挥其抗非小细胞肺癌效应。

Objective Mechanism of action of taraxosterol on non-small cell lung cancer based on in vitro and in vivo models, and its correlation with Wnt/β-catenin signaling were investigated. Methods A549 cells was treated with taraxosterol at different concentrations (0, 5, 10, 20, 40, 80 μmol/L), to optimize the drug concentration selection. A549 cells were divided into control group, taraxosterol (10, 20, and 40 μmol/L), and taraxosterol (40 μmol/L) + CHIR99021 (10 μmol/L) group. Proliferation ability was measured by colony formation assay, apoptosis was analyzed by flow cytometry, and migration and invasion activities were measured by wound healing assay and Transwell chamber assay respectively. Western blotting was used to determine the expression of key molecules in the Wnt/β-catenin signal transduction pathway. Tumor-bearing mouse model was established to further evaluate the in vivo tumor-suppressive effect of taraxosterol and to confirm the critical mediating role of the Wnt/β-catenin pathway in this process. Results Taraxsterol exhibited a significant inhibitory effect on the activity of A549 cells at concentrations ranging from 5 to 80 μmol/L (P < 0.05), with IC50 value of 33.95 μmol/L. Compare with control group, taraxsterol (10, 20, and 40 μmol/L) groups showed significantly lower numbers of clone formation, scratch healing rates, invasive cell counts, and protein expression levels of key molecules in the Wnt pathway (β-catenin, Cyclin D1, c-Myc), as well as a significantly increased degree of GSK-3β phosphorylation, while the cell apoptosis rate significantly increased (P< 0.05). CHIR99021 could reverse the inhibitory effect of taraxsterol on A549 cells. In the animal experiments, compare with model group, the subcutaneous tumor volume and tumor mass of mice in the gypsophila sterol group were significantly reduced (P < 0.05). HE staining results showed that the degree of inflammatory infiltration in the taraxsterol group was significantly reduced, the protein expression levels of β-catenin, c-Myc, Cyclin D1, and GSK-3β phosphorylation were all significantly decreased (P< 0.05). Conclusion Taraxsterol exerts its anti-tumor effects by regulating the Wnt/β-catenin signaling pathway.

R285.5;R286.4