目的 探讨白藜芦醇逆转乳腺癌他莫昔芬（TAM）耐药的作用及分子机制。方法 采用CCK-8法检测TAM作用24、48、72 h对MCF-7亲本细胞及MCF-7/TAM耐药细胞的半数抑制浓度（IC₅₀），鉴定耐药模型；以白藜芦醇（10、20、50 μmol·L^-1）联合TAM干预MCF-7/TAM细胞，检测细胞增殖、凋亡与细胞周期；利用数据非依赖性采集（DIA）定量蛋白质组学筛选差异蛋白，进行基因本体（GO）功能、京都基因与基因组百科全书（KEGG）通路及蛋白质-蛋白质互作（PPI）分析；采用Western blotting验证PI3K/Akt通路关键蛋白表达。结果 MCF-7/TAM细胞各时间点IC₅₀显著高于亲本细胞（P＜ 0.05），成功构建稳定耐药模型。与单给TAM组比较，白藜芦醇联合TAM可显著抑制MCF-7/TAM细胞增殖、提高总凋亡率并将细胞阻滞于G1期（均P＜0.05）。蛋白组学共鉴定到491个显著差异蛋白（上调239个、下调252个），主要富集于细胞周期、细胞凋亡及药物转运过程，涉及原纤维蛋白1（Fbn1）、髓系细胞核分化抗原（Mnda）、高迁移率基团核小体结合域2（Hmgn2）、ATP酶9A（Atp9a）等关键蛋白。GO分析显示，差异蛋白主要富集于分子功能调节因子活性、胞内解剖结构及免疫系统过程。KEGG与PPI分析提示，PI3K/Akt信号通路为核心通路。Western blotting证实，与单给TAM组比较，联合给药可显著下调p-PI3K、p-Akt表达水平（P＜0.05）。结论 白藜芦醇可通过抑制PI3K/Akt信号通路，调控细胞周期、凋亡与药物转运相关蛋白表达，有效逆转MCF-7/TAM细胞对TAM的耐药性。

Objective To investigate the effect and molecular mechanism of resveratrol in reversing tamoxifen resistance in breast cancer. Methods The half-maximal inhibitory concentration (IC₅₀) of TAM on MCF-7 parental cells and MCF-7/TAM resistant cells was determined at 24, 48, and 72 hours using the CCK-8 assay to establish the resistance model. Resveratrol (10, 20, 50 μmol·L^-1) was combined with TAM to treat MCF-7/TAM cells, and cell proliferation, apoptosis, and cell cycle were assessed. Differential proteins were identified via DIA-based quantitative proteomics, followed by gene ontology (GO) functional analysis, KEGG pathway analysis, and protein-protein interaction (PPI) network analysis. Western blotting was used to validate the expression of key proteins in the PI3K/Akt signaling pathway. Results The IC₅₀ values of MCF-7/TAM cells at all time points were significantly higher than those of parental cells (P < 0.05), confirming successful establishment of a stable resistant model. Compared with TAM alone, the combination of resveratrol and TAM significantly suppressed MCF-7/TAM cell proliferation, increased total apoptosis rate, and arrested cells in the G1 phase (all P < 0.05). Proteomic analysis identified 491 differentially expressed proteins (239 upregulated, 252 downregulated), primarily enriched in cell cycle regulation, apoptosis, and drug transport processes, including key proteins such as fibrillin 1 (Fbn1), myeloid nuclear differentiation antigen (Mnda), high-mobility group nucleosome-binding domain 2 (Hmgn2), and ATPase 9A (Atp9a). GO analysis revealed that differentially expressed proteins were mainly involved in molecular function regulation, intracellular anatomical structure, and immune system processes. KEGG and PPI analyses indicated that the PI3K/Akt signaling pathway was the central pathway. Western blotting confirmed that compared with TAM treatment alone, the combination therapy significantly reduced the expression levels of p-PI3K and p-Akt (P < 0.05). Conclusion Resveratrol can effectively reverse tamoxifen resistance in MCF- 7/TAM cells by inhibiting the PI3K/Akt signaling pathway and regulating the expression of proteins related to cell cycle, apoptosis and drug transport.

R965

常州市晋陵药学发展基金项目（应用基础研究指导性）（2025YXHCG12）