目的 探讨槲皮素对人胃癌AGS细胞的抑制作用，分析其调控胃癌增殖的分子机制。方法 采用CCK-8法和平板克隆形成实验检测槲皮素对AGS细胞活力与增殖能力的影响；构建AGS细胞斑马鱼异种移植模型，观察槲皮素的体内抑瘤效果；通过转录组测序，筛选差异表达基因并进行基因本体论（GO）功能注释与京都基因和基因组百科全书（KEGG）通路富集分析；利用透射电镜观察细胞形态变化，共聚焦荧光显微镜检测细胞内脂质活性氧水平；采用分子对接模拟槲皮素与关键蛋白亚精胺/精胺N1-乙酰转移酶1（SAT1）的结合，并通过Western blotting验证SAT1及下游花生四烯酸15-脂氧合酶（ALOX15）蛋白的表达。结果 槲皮素能显著抑制AGS细胞活力与克隆形成（P＜0.01）。在斑马鱼异种移植模型中，槲皮素同样有效抑制了肿瘤细胞的增殖（P＜0.001）。转录组测序分析提示，铁死亡是槲皮素作用的核心通路之一（P＜0.05）。形态学与生化检测证实，槲皮素处理可诱发线粒体皱缩、嵴减少等铁死亡典型超微结构改变，并引起脂质活性氧的显著堆积。分子对接显示槲皮素与SAT1蛋白的C1口袋具有高亲和力（Vina score＝−8.7 kcal/mol）。Western blotting验证表明，槲皮素可上调SAT1及下游ALOX15蛋白表达（P＜0.05、0.001）。结论 槲皮素可诱导胃癌AGS细胞发生铁死亡，可能与上调SAT1/ALOX15通路有关。

Objective To investigate the inhibitory effect of quercetin on human gastric cancer AGS cells, and analyze its molecular mechanism of regulating the proliferation of gastric cancer. Methods The CCK-8 assay and plate colony formation assay were used to detect the effects of quercetin on the viability and proliferation capacity of AGS cells. A zebrafish xenotransplantation model of AGS cells was constructed to observe the antitumor effect of quercetin in vivo. Transcriptome sequencing was performed to screen differentially expressed genes, followed by GO functional annotation and KEGG pathway enrichment analysis. The morphological changes of cells were observed by transmission electron microscope, and the level of lipid reactive oxygen species in cells was detected by confocal fluorescence microscope. Molecular docking was used to simulate the binding of quercetin to the key protein SAT1, and the expression of SAT1 and ALOX15 proteins were validated by Western blotting. Results Quercetin significantly inhibited AGS cell viability and clone formation (P < 0.01). Quercetin also effectively inhibited the proliferation of tumor cells in zebrafish xenotransplantation model (P < 0.001). Transcriptome sequencing analysis suggested that ferroptosis was one of the core pathways of quercetin (P < 0.05). Morphological and biochemical tests confirmed that quercetin could induce the typical ultrastructural changes of ferroptosis, such as mitochondrial shrinkage and crista reduction, and caused significant accumulation of lipid ROS. Molecular docking showed that quercetin had a high affinity with C1 pocket of SAT1 protein (Vina score=-8.7 kcal/mol).Western blotting analysis verified that quercetin could upregulate the protein expression of both SAT1 and its downstream protein ALOX15 (P < 0.05, 0.001). Conclusion Quercetin could induce ferroptosis in gastric cancer AGS cells, which might be associated with the upregulating SAT1/ALOX15 pathway.

R286.5

国家自然科学基金资助项目(82374410); 国家中管局中医脾胃病学高水平重点学科建设项目(zyyzdxk-202375); 江苏省中医药管理局科技项目(JD2022SZ02); 江苏省中医院科技项目(Y2020CX68)